{
  "pet28a": {
    "id": "pet28a",
    "name": "pET-28a(+)",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 5369,
    "promoter": "T7 lac",
    "induction": "IPTG (0.1–1 mM)",
    "selection": [
      "kanamycin (50 µg/mL)"
    ],
    "copy": "medium",
    "fusion_tag": {
      "n_term": "MGSSHHHHHHSSGLVPRGSHM (His6 + thrombin)",
      "c_term": "LEHHHHHH (His6)"
    },
    "mcs_enzymes": [
      "NcoI",
      "NheI",
      "NdeI",
      "BamHI",
      "EcoRV",
      "EcoRI",
      "SacI",
      "SalI",
      "HindIII",
      "NotI",
      "XhoI"
    ],
    "flanking_5p_max": "AAGGAGATATACCATGGCT",
    "flanking_3p_max": "CTCGAGCACCACCACCACCACCACTGAGATCC",
    "addgene": "69864-3",
    "supplier": "Sigma-Aldrich / Novagen",
    "notes": "Workhorse bacterial expression vector. T7 promoter requires DE3-lysogen host (BL21(DE3), Rosetta, etc.)."
  },
  "pet21a": {
    "id": "pet21a",
    "name": "pET-21a(+)",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 5443,
    "promoter": "T7 lac",
    "induction": "IPTG",
    "selection": [
      "ampicillin (100 µg/mL)"
    ],
    "copy": "medium",
    "fusion_tag": {
      "n_term": "T7-tag (MASMTGGQQMG)",
      "c_term": "LEHHHHHH (His6)"
    },
    "mcs_enzymes": [
      "NdeI",
      "NheI",
      "BamHI",
      "EcoRI",
      "SacI",
      "SalI",
      "HindIII",
      "NotI",
      "EagI",
      "XhoI"
    ],
    "flanking_5p_max": "AAGGAGATATACATATG",
    "flanking_3p_max": "CTCGAGCACCACCACCACCACCACTGA",
    "addgene": "69740",
    "supplier": "Sigma / Novagen",
    "notes": "Like pET-28a but amp-resistant and no N-term His tag."
  },
  "pet22b": {
    "id": "pet22b",
    "name": "pET-22b(+)",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 5493,
    "promoter": "T7 lac",
    "induction": "IPTG",
    "selection": [
      "ampicillin"
    ],
    "copy": "medium",
    "fusion_tag": {
      "n_term": "pelB signal (periplasmic export)",
      "c_term": "LEHHHHHH"
    },
    "mcs_enzymes": [
      "NdeI",
      "NcoI",
      "BamHI",
      "EcoRI",
      "SacI",
      "SalI",
      "HindIII",
      "NotI",
      "XhoI"
    ],
    "flanking_5p_max": "AAGGAGATATACATATG",
    "flanking_3p_max": "CTCGAGCACCACCACCACCACCACTGA",
    "notes": "pelB signal peptide directs protein to the periplasm — useful for disulfide-containing or toxic proteins."
  },
  "pet32a": {
    "id": "pet32a",
    "name": "pET-32a(+)",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 5900,
    "promoter": "T7 lac",
    "induction": "IPTG",
    "selection": [
      "ampicillin"
    ],
    "copy": "medium",
    "fusion_tag": {
      "n_term": "Trx + His6 + S-tag + thrombin",
      "c_term": "His6"
    },
    "mcs_enzymes": [
      "KpnI",
      "BamHI",
      "EcoRI",
      "EcoRV",
      "AvaI",
      "HindIII",
      "SalI",
      "XhoI",
      "NotI"
    ],
    "flanking_5p_max": "GGTACC",
    "flanking_3p_max": "CTCGAG",
    "notes": "N-terminal thioredoxin fusion enhances solubility of difficult proteins."
  },
  "pgex_6p_1": {
    "id": "pgex_6p_1",
    "name": "pGEX-6P-1",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 4984,
    "promoter": "tac",
    "induction": "IPTG",
    "selection": [
      "ampicillin"
    ],
    "copy": "medium",
    "fusion_tag": {
      "n_term": "GST + PreScission protease site (LEVLFQ↓GP)",
      "c_term": "none"
    },
    "mcs_enzymes": [
      "BamHI",
      "EcoRI",
      "SmaI",
      "SalI",
      "XhoI",
      "NotI"
    ],
    "flanking_5p_max": "CTGGAAGTTCTGTTCCAGGGGCCC",
    "flanking_3p_max": "TAATAA",
    "supplier": "Cytiva / GE Healthcare",
    "notes": "N-terminal GST fusion (~26 kDa). Purify on glutathione resin, cleave with PreScission to release tag."
  },
  "pmal_c5x": {
    "id": "pmal_c5x",
    "name": "pMAL-c5X",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 5677,
    "promoter": "tac",
    "induction": "IPTG",
    "selection": [
      "ampicillin"
    ],
    "copy": "medium",
    "fusion_tag": {
      "n_term": "MBP + Factor Xa site (IEGR↓)",
      "c_term": "none"
    },
    "mcs_enzymes": [
      "NdeI",
      "EcoRI",
      "BamHI",
      "XbaI",
      "SalI",
      "PstI",
      "HindIII"
    ],
    "flanking_5p_max": "GAAGGTAGGTCATATG",
    "flanking_3p_max": "AAGCTT",
    "supplier": "NEB",
    "notes": "N-terminal MBP (~42 kDa) — strong solubility enhancer for difficult proteins."
  },
  "puc19": {
    "id": "puc19",
    "name": "pUC19",
    "host": "e_coli",
    "category": "general_cloning",
    "size_bp": 2686,
    "promoter": "lac (for α-complementation)",
    "induction": "none",
    "selection": [
      "ampicillin"
    ],
    "copy": "high",
    "fusion_tag": {
      "n_term": "none",
      "c_term": "none"
    },
    "mcs_enzymes": [
      "EcoRI",
      "SacI",
      "KpnI",
      "SmaI",
      "BamHI",
      "XbaI",
      "SalI",
      "AccI",
      "HincII",
      "PstI",
      "SphI",
      "HindIII"
    ],
    "flanking_5p_max": "GAATTC",
    "flanking_3p_max": "AAGCTT",
    "notes": "High-copy cloning vector. Blue-white screening via lacZα. No expression machinery — for shuttling DNA only."
  },
  "pbluescript": {
    "id": "pbluescript",
    "name": "pBluescript II SK(+)",
    "host": "e_coli",
    "category": "general_cloning",
    "size_bp": 2961,
    "promoter": "T7, T3 (for in vitro RNA)",
    "induction": "n/a",
    "selection": [
      "ampicillin"
    ],
    "copy": "high",
    "fusion_tag": {
      "n_term": "none",
      "c_term": "none"
    },
    "mcs_enzymes": [
      "KpnI",
      "ApaI",
      "XhoI",
      "SalI",
      "AccI",
      "HincII",
      "ClaI",
      "HindIII",
      "EcoRV",
      "EcoRI",
      "PstI",
      "SmaI",
      "BamHI",
      "SpeI",
      "XbaI",
      "NotI",
      "SacII",
      "SacI"
    ],
    "flanking_5p_max": "GGTACC",
    "flanking_3p_max": "GAGCTC",
    "supplier": "Agilent / Stratagene",
    "notes": "Versatile cloning vector with flanking T7 + T3 promoters for in vitro RNA transcription from either strand."
  },
  "pcdf_duet": {
    "id": "pcdf_duet",
    "name": "pCDFDuet-1",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 3781,
    "promoter": "two T7 lac (dual MCS)",
    "induction": "IPTG",
    "selection": [
      "streptomycin (50 µg/mL)"
    ],
    "copy": "low (CDF origin)",
    "fusion_tag": {
      "n_term": "optional His6 (MCS1)",
      "c_term": "optional S-tag (MCS2)"
    },
    "mcs_enzymes": [
      "NcoI",
      "NdeI",
      "BamHI",
      "EcoRI",
      "SacI",
      "NotI",
      "AvrII",
      "PstI",
      "HindIII",
      "XhoI",
      "BglII",
      "KpnI",
      "PacI"
    ],
    "flanking_5p_max": "AAGGAGATATACCATGG",
    "flanking_3p_max": "CTCGAG",
    "notes": "Dual-expression vector for co-expression with pET or pACYC partners (different antibiotics, different origins)."
  },
  "pcdna3_1": {
    "id": "pcdna3_1",
    "name": "pcDNA3.1(+)",
    "host": "mammalian",
    "category": "mammalian_expression",
    "size_bp": 5428,
    "promoter": "CMV",
    "induction": "constitutive",
    "selection": [
      "ampicillin (E. coli)",
      "G418/neomycin (mammalian)"
    ],
    "copy": "episomal in mammalian",
    "fusion_tag": {
      "n_term": "none (optional)",
      "c_term": "none"
    },
    "mcs_enzymes": [
      "NheI",
      "AflII",
      "HindIII",
      "KpnI",
      "BamHI",
      "BstXI",
      "EcoRI",
      "EcoRV",
      "NotI",
      "XhoI",
      "XbaI",
      "ApaI"
    ],
    "flanking_5p_max": "GCTAGC",
    "flanking_3p_max": "CTCGAGTCTAGAGG",
    "supplier": "Invitrogen / Thermo Fisher",
    "notes": "Standard CMV-driven mammalian expression vector. Includes BGH polyA and SV40 ori."
  },
  "pcdna3_4": {
    "id": "pcdna3_4",
    "name": "pcDNA3.4",
    "host": "mammalian",
    "category": "mammalian_expression",
    "size_bp": 5810,
    "promoter": "CMV",
    "induction": "constitutive",
    "selection": [
      "ampicillin",
      "G418"
    ],
    "copy": "episomal",
    "mcs_enzymes": [
      "NotI",
      "NheI",
      "AscI",
      "HindIII",
      "KpnI",
      "EcoRI",
      "BamHI",
      "EcoRV",
      "XhoI"
    ],
    "flanking_5p_max": "GCTAGC",
    "flanking_3p_max": "CTCGAG",
    "supplier": "Thermo Fisher",
    "notes": "Optimized for CHO/HEK293 transient expression. Used for therapeutic antibody / protein production."
  },
  "pegfp_n1": {
    "id": "pegfp_n1",
    "name": "pEGFP-N1",
    "host": "mammalian",
    "category": "mammalian_expression",
    "size_bp": 4733,
    "promoter": "CMV",
    "induction": "constitutive",
    "selection": [
      "kanamycin (E. coli)",
      "G418 (mammalian)"
    ],
    "fusion_tag": {
      "n_term": "none",
      "c_term": "EGFP"
    },
    "mcs_enzymes": [
      "NheI",
      "AgeI",
      "EcoRI",
      "PstI",
      "SalI",
      "AccI",
      "XhoI",
      "BamHI",
      "HindIII",
      "SacII",
      "KpnI",
      "XmaI",
      "SmaI"
    ],
    "flanking_5p_max": "GCTAGC",
    "flanking_3p_max": "GGTACCGCGGGCCCGGGATCC",
    "supplier": "Clontech / Takara",
    "notes": "C-terminal EGFP fusion for live-cell imaging. Insert in frame at the MCS, just before the EGFP ORF."
  },
  "pyes2": {
    "id": "pyes2",
    "name": "pYES2",
    "host": "yeast",
    "category": "yeast_expression",
    "size_bp": 5856,
    "promoter": "GAL1 (galactose-inducible)",
    "induction": "galactose 2%",
    "selection": [
      "ampicillin (E. coli)",
      "URA3 (yeast — ura3- strain required)"
    ],
    "mcs_enzymes": [
      "KpnI",
      "BamHI",
      "BstXI",
      "EcoRI",
      "NotI",
      "XhoI",
      "SphI",
      "XbaI"
    ],
    "flanking_5p_max": "GGTACC",
    "flanking_3p_max": "TCTAGA",
    "supplier": "Invitrogen",
    "notes": "S. cerevisiae GAL1-driven expression. Tight off-state on glucose; strong induction on galactose."
  },
  "ppicz": {
    "id": "ppicz",
    "name": "pPICZ-α",
    "host": "pichia",
    "category": "yeast_expression",
    "size_bp": 3593,
    "promoter": "AOX1 (methanol-inducible)",
    "induction": "methanol",
    "selection": [
      "Zeocin (E. coli + yeast)"
    ],
    "fusion_tag": {
      "n_term": "α-factor signal (secreted)",
      "c_term": "optional Myc + His6"
    },
    "mcs_enzymes": [
      "EcoRI",
      "PmlI",
      "SfiI",
      "BstBI",
      "AvrII",
      "PvuII",
      "NotI",
      "SacII",
      "XbaI"
    ],
    "flanking_5p_max": "GAATTC",
    "flanking_3p_max": "TCTAGA",
    "supplier": "Invitrogen",
    "notes": "Pichia pastoris methanol-inducible expression with secretion signal. High protein yields."
  },
  "pet15b": {
    "id": "pet15b",
    "name": "pET-15b",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 5708,
    "promoter": "T7 lac",
    "induction": "IPTG",
    "selection": [
      "ampicillin"
    ],
    "copy": "medium",
    "fusion_tag": {
      "n_term": "MGSSHHHHHHSSGLVPRGSHM (His6 + thrombin)",
      "c_term": "none"
    },
    "mcs_enzymes": [
      "NdeI",
      "XhoI",
      "BamHI"
    ],
    "flanking_5p_max": "AAGGAGATATACATATG",
    "flanking_3p_max": "GGATCCTGA",
    "supplier": "Sigma-Aldrich / Novagen",
    "notes": "Compact pET vector with N-terminal His6 + thrombin cleavage. Smaller MCS than pET-28a."
  },
  "pet30a": {
    "id": "pet30a",
    "name": "pET-30a(+)",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 5422,
    "promoter": "T7 lac",
    "induction": "IPTG",
    "selection": [
      "kanamycin"
    ],
    "copy": "medium",
    "fusion_tag": {
      "n_term": "His6 + thrombin + S-tag + enterokinase",
      "c_term": "His6"
    },
    "mcs_enzymes": [
      "NcoI",
      "NdeI",
      "NheI",
      "BamHI",
      "EcoRI",
      "SacI",
      "SalI",
      "HindIII",
      "NotI",
      "XhoI"
    ],
    "flanking_5p_max": "AAGGAGATATACATATG",
    "flanking_3p_max": "CTCGAGCACCACCACCACCACCAC",
    "supplier": "Sigma-Aldrich / Novagen",
    "notes": "Like pET-28a but with N-terminal S-tag and enterokinase site for tag removal."
  },
  "pet3a": {
    "id": "pet3a",
    "name": "pET-3a",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 4640,
    "promoter": "T7",
    "induction": "IPTG",
    "selection": [
      "ampicillin"
    ],
    "copy": "medium",
    "fusion_tag": {
      "n_term": "T7 gene 10 leader (11 aa)",
      "c_term": "none"
    },
    "mcs_enzymes": [
      "NdeI",
      "BamHI"
    ],
    "flanking_5p_max": "AAGGAGATATACATATG",
    "flanking_3p_max": "GGATCCTGA",
    "notes": "Minimal pET — tagless expression for downstream crystallography or activity studies."
  },
  "pbad_hisa": {
    "id": "pbad_hisa",
    "name": "pBAD/HisA",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 4102,
    "promoter": "araBAD (arabinose-inducible)",
    "induction": "L-arabinose (0.0002–0.2%)",
    "selection": [
      "ampicillin"
    ],
    "copy": "medium",
    "fusion_tag": {
      "n_term": "His6 + Xpress epitope + enterokinase",
      "c_term": "none"
    },
    "mcs_enzymes": [
      "NcoI",
      "PstI",
      "KpnI",
      "EcoRI",
      "BglII",
      "XhoI",
      "EcoRV",
      "NheI",
      "HindIII"
    ],
    "flanking_5p_max": "CCATGG",
    "flanking_3p_max": "AAGCTT",
    "supplier": "Invitrogen",
    "notes": "Tightly tunable expression via arabinose titration. Best for toxic proteins where leaky expression kills cells."
  },
  "pcold_i": {
    "id": "pcold_i",
    "name": "pCold I",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 4407,
    "promoter": "cspA (cold-shock)",
    "induction": "15 °C cold shock + IPTG",
    "selection": [
      "ampicillin"
    ],
    "copy": "medium",
    "fusion_tag": {
      "n_term": "His6",
      "c_term": "none"
    },
    "mcs_enzymes": [
      "NdeI",
      "SacI",
      "KpnI",
      "XhoI",
      "BamHI",
      "EcoRI",
      "HindIII"
    ],
    "flanking_5p_max": "CATATG",
    "flanking_3p_max": "GGATCC",
    "supplier": "Takara Bio",
    "notes": "Cold-shock induction at 15 °C improves folding of difficult proteins. Most host proteases inactive at this temperature."
  },
  "ptrc99a": {
    "id": "ptrc99a",
    "name": "pTrc99A",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 4176,
    "promoter": "trc (trp/lac hybrid)",
    "induction": "IPTG",
    "selection": [
      "ampicillin"
    ],
    "copy": "medium",
    "fusion_tag": {
      "n_term": "none",
      "c_term": "none"
    },
    "mcs_enzymes": [
      "NcoI",
      "EcoRI",
      "SacI",
      "KpnI",
      "SmaI",
      "BamHI",
      "XbaI",
      "SalI",
      "AccI",
      "HincII",
      "PstI",
      "SphI",
      "HindIII"
    ],
    "flanking_5p_max": "CCATGG",
    "flanking_3p_max": "AAGCTT",
    "supplier": "Pharmacia / Cytiva",
    "notes": "Trc promoter is stronger than lac, weaker than T7 — does not require DE3 strain. Tagless expression."
  },
  "pet_duet": {
    "id": "pet_duet",
    "name": "pETDuet-1",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 5420,
    "promoter": "two T7 lac (dual MCS)",
    "induction": "IPTG",
    "selection": [
      "ampicillin"
    ],
    "copy": "medium (ColE1)",
    "fusion_tag": {
      "n_term": "His6 (MCS1)",
      "c_term": "S-tag (MCS2)"
    },
    "mcs_enzymes": [
      "NcoI",
      "BamHI",
      "EcoRI",
      "SacI",
      "NotI",
      "SalI",
      "HindIII",
      "PstI",
      "KpnI",
      "AvrII",
      "XhoI"
    ],
    "flanking_5p_max": "CCATGG",
    "flanking_3p_max": "CTCGAG",
    "notes": "Dual-expression for co-expressing two proteins from one plasmid. Compatible with pCDFDuet, pACYCDuet, pRSFDuet."
  },
  "pacyc_duet": {
    "id": "pacyc_duet",
    "name": "pACYCDuet-1",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 4008,
    "promoter": "two T7 lac (dual MCS)",
    "induction": "IPTG",
    "selection": [
      "chloramphenicol (34 µg/mL)"
    ],
    "copy": "low (p15A ori)",
    "fusion_tag": {
      "n_term": "His6 (MCS1)",
      "c_term": "S-tag (MCS2)"
    },
    "mcs_enzymes": [
      "NcoI",
      "BamHI",
      "EcoRI",
      "SacI",
      "NotI",
      "SalI",
      "HindIII",
      "PstI",
      "KpnI",
      "AvrII",
      "XhoI"
    ],
    "flanking_5p_max": "CCATGG",
    "flanking_3p_max": "CTCGAG",
    "notes": "Compatible with pETDuet/pCDFDuet/pRSFDuet for up to 8-protein co-expression. Different antibiotic, different origin."
  },
  "pcoldtf": {
    "id": "pcoldtf",
    "name": "pCold TF",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 5821,
    "promoter": "cspA",
    "induction": "15 °C cold shock + IPTG",
    "selection": [
      "ampicillin"
    ],
    "copy": "medium",
    "fusion_tag": {
      "n_term": "Trigger Factor (~48 kDa) + His6 + thrombin",
      "c_term": "none"
    },
    "mcs_enzymes": [
      "KpnI",
      "SacI",
      "BamHI",
      "EcoRI",
      "EcoRV",
      "HindIII",
      "SalI",
      "PstI",
      "XbaI",
      "XhoI"
    ],
    "flanking_5p_max": "GGTACC",
    "flanking_3p_max": "CTCGAG",
    "supplier": "Takara Bio",
    "notes": "Cold-shock + Trigger Factor chaperone fusion — strongest folding-rescue option for aggregation-prone proteins."
  },
  "psb1c3": {
    "id": "psb1c3",
    "name": "pSB1C3",
    "host": "e_coli",
    "category": "general_cloning",
    "size_bp": 2070,
    "promoter": "none (BioBrick assembly)",
    "induction": "n/a",
    "selection": [
      "chloramphenicol"
    ],
    "copy": "high (pUC ori)",
    "fusion_tag": {
      "n_term": "none",
      "c_term": "none"
    },
    "mcs_enzymes": [
      "EcoRI",
      "XbaI",
      "SpeI",
      "PstI",
      "NotI"
    ],
    "flanking_5p_max": "GAATTCGCGGCCGCTTCTAGAG",
    "flanking_3p_max": "TACTAGTAGCGGCCGCTGCAG",
    "notes": "iGEM standard BioBrick assembly vector (RFC 10). Part insertion between BioBrick prefix and suffix."
  },
  "pcdna3_1_neg": {
    "id": "pcdna3_1_neg",
    "name": "pcDNA3.1(-)",
    "host": "mammalian",
    "category": "mammalian_expression",
    "size_bp": 5427,
    "promoter": "CMV",
    "induction": "constitutive",
    "selection": [
      "ampicillin",
      "G418/neomycin"
    ],
    "copy": "episomal",
    "mcs_enzymes": [
      "BamHI",
      "EcoRI",
      "EcoRV",
      "BstXI",
      "NotI",
      "XhoI",
      "XbaI",
      "ApaI",
      "KpnI",
      "HindIII",
      "AflII",
      "NheI"
    ],
    "flanking_5p_max": "GGATCC",
    "flanking_3p_max": "GCTAGC",
    "supplier": "Invitrogen",
    "notes": "Reverse MCS orientation of pcDNA3.1(+). Use when cloning in opposite reading direction is needed."
  },
  "pcdna4_to": {
    "id": "pcdna4_to",
    "name": "pcDNA4/TO",
    "host": "mammalian",
    "category": "mammalian_expression",
    "size_bp": 5078,
    "promoter": "CMV + Tet operator (2× TetO2)",
    "induction": "tetracycline / doxycycline",
    "selection": [
      "ampicillin",
      "Zeocin"
    ],
    "copy": "episomal",
    "mcs_enzymes": [
      "HindIII",
      "KpnI",
      "BamHI",
      "EcoRI",
      "EcoRV",
      "NotI",
      "XhoI",
      "XbaI",
      "ApaI",
      "NheI"
    ],
    "flanking_5p_max": "AAGCTT",
    "flanking_3p_max": "CTCGAG",
    "supplier": "Invitrogen",
    "notes": "Tet-on inducible mammalian expression. Use with T-REx host cell line (stable TetR expression)."
  },
  "pcdh_cmv": {
    "id": "pcdh_cmv",
    "name": "pCDH-CMV-MCS-EF1-Puro",
    "host": "mammalian",
    "category": "lentiviral",
    "size_bp": 7385,
    "promoter": "CMV (transgene) + EF1α (Puro selection)",
    "induction": "constitutive",
    "selection": [
      "ampicillin (E. coli)",
      "puromycin (mammalian)"
    ],
    "copy": "integrated",
    "mcs_enzymes": [
      "EcoRI",
      "BamHI",
      "NheI",
      "XbaI",
      "NotI",
      "SwaI",
      "EcoRV",
      "SalI"
    ],
    "flanking_5p_max": "GAATTC",
    "flanking_3p_max": "GCGGCCGC",
    "supplier": "System Biosciences (SBI)",
    "notes": "3rd-gen lentiviral transfer vector. Co-transfect with pPAX2 + pMD2.G to produce virus. Integrates into host genome."
  },
  "pires2_egfp": {
    "id": "pires2_egfp",
    "name": "pIRES2-EGFP",
    "host": "mammalian",
    "category": "mammalian_expression",
    "size_bp": 5298,
    "promoter": "CMV",
    "induction": "constitutive",
    "selection": [
      "kanamycin (E. coli)",
      "G418 (mammalian)"
    ],
    "copy": "episomal",
    "fusion_tag": {
      "n_term": "none (bicistronic — separate EGFP via IRES)",
      "c_term": "none"
    },
    "mcs_enzymes": [
      "NheI",
      "EcoRI",
      "PstI",
      "SalI",
      "AccI",
      "XhoI",
      "BamHI",
      "BstXI",
      "EcoRV",
      "NotI",
      "XbaI",
      "SacII"
    ],
    "flanking_5p_max": "GCTAGC",
    "flanking_3p_max": "TCTAGA",
    "supplier": "Clontech / Takara",
    "notes": "Bicistronic: gene-of-interest + IRES + EGFP. EGFP marks transfected cells without fusing to your protein."
  },
  "ptre3g": {
    "id": "ptre3g",
    "name": "pTRE3G",
    "host": "mammalian",
    "category": "mammalian_expression",
    "size_bp": 3187,
    "promoter": "PTRE3G (Tet-On 3G inducible)",
    "induction": "doxycycline (1–1000 ng/mL)",
    "selection": [
      "ampicillin"
    ],
    "copy": "episomal",
    "mcs_enzymes": [
      "BamHI",
      "MluI",
      "SalI",
      "NheI",
      "EcoRI",
      "NotI",
      "PvuI",
      "EcoRV",
      "HindIII"
    ],
    "flanking_5p_max": "GGATCC",
    "flanking_3p_max": "AAGCTT",
    "supplier": "Clontech / Takara",
    "notes": "Tightest tet-inducible system. Requires co-expression of Tet-On 3G transactivator (pCMV-Tet3G)."
  },
  "pcmv_ha": {
    "id": "pcmv_ha",
    "name": "pCMV-HA",
    "host": "mammalian",
    "category": "mammalian_expression",
    "size_bp": 3787,
    "promoter": "CMV",
    "induction": "constitutive",
    "selection": [
      "ampicillin"
    ],
    "copy": "episomal",
    "fusion_tag": {
      "n_term": "HA epitope (YPYDVPDYA)",
      "c_term": "none"
    },
    "mcs_enzymes": [
      "SalI",
      "AccI",
      "XbaI",
      "NotI",
      "EcoRI",
      "BglII",
      "KpnI",
      "SacI"
    ],
    "flanking_5p_max": "GTCGAC",
    "flanking_3p_max": "GAGCTC",
    "supplier": "Clontech",
    "notes": "N-terminal HA tag for immunoprecipitation and detection. Anti-HA antibodies are well-validated."
  },
  "p3xflag_cmv10": {
    "id": "p3xflag_cmv10",
    "name": "p3xFLAG-CMV-10",
    "host": "mammalian",
    "category": "mammalian_expression",
    "size_bp": 6259,
    "promoter": "CMV",
    "induction": "constitutive",
    "selection": [
      "ampicillin"
    ],
    "copy": "episomal",
    "fusion_tag": {
      "n_term": "3×FLAG (DYKDHDGDYKDHDIDYKDDDDK) + preprotrypsin signal",
      "c_term": "none"
    },
    "mcs_enzymes": [
      "NotI",
      "HindIII",
      "BglII",
      "EcoRI",
      "EcoRV",
      "BamHI",
      "SalI",
      "KpnI",
      "XbaI",
      "XhoI"
    ],
    "flanking_5p_max": "AAGCTT",
    "flanking_3p_max": "CTCGAG",
    "supplier": "Sigma-Aldrich",
    "notes": "Secreted 3×FLAG fusion. M2 anti-FLAG resin allows highly specific purification + competitive elution with FLAG peptide."
  },
  "plko1": {
    "id": "plko1",
    "name": "pLKO.1-puro",
    "host": "mammalian",
    "category": "lentiviral",
    "size_bp": 7032,
    "promoter": "U6 (shRNA) + PGK (Puro)",
    "induction": "constitutive",
    "selection": [
      "ampicillin (E. coli)",
      "puromycin (mammalian)"
    ],
    "copy": "integrated",
    "mcs_enzymes": [
      "AgeI",
      "EcoRI"
    ],
    "flanking_5p_max": "ACCGG",
    "flanking_3p_max": "CTTTTTG",
    "addgene": "10878",
    "notes": "Standard lentiviral shRNA knockdown vector. Clone hairpin between AgeI and EcoRI behind U6 promoter."
  },
  "lenticrispr_v2": {
    "id": "lenticrispr_v2",
    "name": "lentiCRISPR v2",
    "host": "mammalian",
    "category": "lentiviral",
    "size_bp": 13044,
    "promoter": "EF1α-short (Cas9) + U6 (sgRNA)",
    "induction": "constitutive",
    "selection": [
      "ampicillin",
      "puromycin"
    ],
    "copy": "integrated",
    "mcs_enzymes": [
      "BsmBI"
    ],
    "flanking_5p_max": "CACCG",
    "flanking_3p_max": "GTTT",
    "addgene": "52961",
    "notes": "Single-plasmid CRISPR/Cas9 + sgRNA + Puro. Golden Gate BsmBI sgRNA cloning. Zhang lab."
  },
  "px330": {
    "id": "px330",
    "name": "pX330-U6-Chimeric_BB-CBh-hSpCas9",
    "host": "mammalian",
    "category": "mammalian_expression",
    "size_bp": 8484,
    "promoter": "CBh (Cas9) + U6 (sgRNA)",
    "induction": "constitutive",
    "selection": [
      "ampicillin"
    ],
    "copy": "episomal",
    "mcs_enzymes": [
      "BbsI"
    ],
    "flanking_5p_max": "CACCG",
    "flanking_3p_max": "GTTT",
    "addgene": "42230",
    "notes": "Non-viral all-in-one CRISPR/Cas9 + sgRNA expression. BbsI Golden Gate sgRNA cloning. Zhang lab."
  },
  "pgbkt7": {
    "id": "pgbkt7",
    "name": "pGBKT7",
    "host": "yeast",
    "category": "yeast_expression",
    "size_bp": 7301,
    "promoter": "ADH1",
    "induction": "constitutive",
    "selection": [
      "kanamycin (E. coli)",
      "TRP1 (yeast)"
    ],
    "copy": "low",
    "fusion_tag": {
      "n_term": "GAL4 DNA-binding domain (1–147) + Myc",
      "c_term": "none"
    },
    "mcs_enzymes": [
      "NdeI",
      "NcoI",
      "SfiI",
      "EcoRI",
      "SmaI",
      "BamHI",
      "SalI",
      "PstI"
    ],
    "flanking_5p_max": "CATATG",
    "flanking_3p_max": "AAGCTT",
    "supplier": "Clontech / Takara",
    "notes": "Yeast two-hybrid \"bait\" vector. Co-transform with pGADT7 prey in AH109/Y2HGold to screen interactions."
  },
  "pgadt7": {
    "id": "pgadt7",
    "name": "pGADT7",
    "host": "yeast",
    "category": "yeast_expression",
    "size_bp": 8000,
    "promoter": "ADH1",
    "induction": "constitutive",
    "selection": [
      "ampicillin (E. coli)",
      "LEU2 (yeast)"
    ],
    "copy": "high",
    "fusion_tag": {
      "n_term": "GAL4 activation domain (768–881) + HA",
      "c_term": "none"
    },
    "mcs_enzymes": [
      "NdeI",
      "SfiI",
      "NcoI",
      "EcoRI",
      "SmaI",
      "BamHI",
      "SacI",
      "XhoI"
    ],
    "flanking_5p_max": "CATATG",
    "flanking_3p_max": "CTCGAG",
    "supplier": "Clontech / Takara",
    "notes": "Yeast two-hybrid \"prey\" vector. Complements pGBKT7 — partner in Matchmaker Y2H system."
  },
  "pyd1": {
    "id": "pyd1",
    "name": "pYD1",
    "host": "yeast",
    "category": "yeast_expression",
    "size_bp": 5024,
    "promoter": "GAL1",
    "induction": "galactose 2%",
    "selection": [
      "ampicillin (E. coli)",
      "TRP1 (yeast)"
    ],
    "copy": "medium",
    "fusion_tag": {
      "n_term": "AGA2 signal + Aga2p (surface anchor) + HA",
      "c_term": "Myc"
    },
    "mcs_enzymes": [
      "NheI",
      "BamHI",
      "EcoRI",
      "NotI",
      "XhoI"
    ],
    "flanking_5p_max": "GCTAGC",
    "flanking_3p_max": "CTCGAG",
    "supplier": "Invitrogen",
    "notes": "Yeast surface display: protein displayed on cell wall via Aga1-Aga2 disulfide. Used for directed evolution by FACS."
  },
  "pcambia1300": {
    "id": "pcambia1300",
    "name": "pCAMBIA1300",
    "host": "plant",
    "category": "plant_expression",
    "size_bp": 8958,
    "promoter": "CaMV 35S (transgene cassette optional)",
    "induction": "n/a",
    "selection": [
      "kanamycin (bacteria)",
      "hygromycin (plant)"
    ],
    "copy": "low",
    "mcs_enzymes": [
      "EcoRI",
      "KpnI",
      "XbaI",
      "SalI",
      "PstI",
      "HindIII",
      "BamHI",
      "SacI",
      "XhoI"
    ],
    "flanking_5p_max": "GAATTC",
    "flanking_3p_max": "AAGCTT",
    "supplier": "CAMBIA",
    "notes": "Agrobacterium-mediated plant transformation binary vector. T-DNA borders flank the MCS."
  },
  "pbi121": {
    "id": "pbi121",
    "name": "pBI121",
    "host": "plant",
    "category": "plant_expression",
    "size_bp": 14758,
    "promoter": "CaMV 35S + NOS",
    "induction": "constitutive",
    "selection": [
      "kanamycin (bacteria + plant)"
    ],
    "copy": "low",
    "mcs_enzymes": [
      "HindIII",
      "XbaI",
      "SmaI",
      "BamHI",
      "SacI",
      "EcoRI"
    ],
    "flanking_5p_max": "TCTAGA",
    "flanking_3p_max": "GAGCTC",
    "supplier": "Clontech",
    "notes": "Classic Agrobacterium binary vector. GUS reporter in MCS — replace with your gene via XbaI + SacI."
  },
  "pgem_t_easy": {
    "id": "pgem_t_easy",
    "name": "pGEM-T Easy",
    "host": "e_coli",
    "category": "general_cloning",
    "size_bp": 3015,
    "promoter": "T7 + SP6 (in vitro RNA)",
    "induction": "n/a",
    "selection": [
      "ampicillin"
    ],
    "copy": "high",
    "mcs_enzymes": [
      "EcoRI",
      "BstZI",
      "NotI",
      "NdeI",
      "SacI",
      "SacII",
      "SpeI",
      "NsiI"
    ],
    "flanking_5p_max": "GAATTC",
    "flanking_3p_max": "GAATTC",
    "supplier": "Promega",
    "notes": "TA cloning vector with single 3′-T overhangs for direct ligation of Taq-amplified PCR products. Blue-white screening."
  },
  "pjet12": {
    "id": "pjet12",
    "name": "pJET1.2/blunt",
    "host": "e_coli",
    "category": "general_cloning",
    "size_bp": 2974,
    "promoter": "T7 (in vitro)",
    "induction": "n/a",
    "selection": [
      "ampicillin"
    ],
    "copy": "high",
    "mcs_enzymes": [
      "EcoRV (blunt)",
      "BglII",
      "XhoI",
      "XbaI",
      "BamHI",
      "NotI",
      "NcoI",
      "HindIII"
    ],
    "flanking_5p_max": "AGATCT",
    "flanking_3p_max": "AAGCTT",
    "supplier": "Thermo Fisher",
    "notes": "Blunt-end PCR cloning with positive selection (lethal eco47IR disrupted by insert). No blue-white needed."
  },
  "paav_mcs": {
    "id": "paav_mcs",
    "name": "pAAV-MCS",
    "host": "mammalian",
    "category": "aav",
    "size_bp": 5311,
    "promoter": "CMV",
    "induction": "constitutive",
    "selection": [
      "ampicillin"
    ],
    "copy": "packaged into AAV particles",
    "mcs_enzymes": [
      "EcoRI",
      "BamHI",
      "SalI",
      "PstI",
      "HindIII",
      "NotI",
      "XhoI",
      "XbaI"
    ],
    "flanking_5p_max": "GAATTC",
    "flanking_3p_max": "AAGCTT",
    "supplier": "Agilent / Stratagene",
    "notes": "AAV transfer vector. Co-transfect with pAAV-RC (rep/cap) + pHelper into HEK293 to produce virus. Insert capacity ~4.7 kb between ITRs."
  },
  "pfastbac1": {
    "id": "pfastbac1",
    "name": "pFastBac1",
    "host": "insect",
    "category": "baculovirus",
    "size_bp": 4776,
    "promoter": "polyhedrin (PH)",
    "induction": "baculovirus infection",
    "selection": [
      "ampicillin",
      "gentamicin (bacmid)"
    ],
    "copy": "medium",
    "mcs_enzymes": [
      "BamHI",
      "EcoRI",
      "SacI",
      "KpnI",
      "SmaI",
      "NotI",
      "XhoI",
      "XbaI",
      "SphI",
      "HindIII"
    ],
    "flanking_5p_max": "GGATCC",
    "flanking_3p_max": "AAGCTT",
    "supplier": "Invitrogen / Thermo Fisher",
    "notes": "Bac-to-Bac baculovirus expression. Transpose into DH10Bac bacmid, transfect Sf9/Sf21 cells. Strong PH-driven late expression."
  },
  "pfastbac_hta": {
    "id": "pfastbac_hta",
    "name": "pFastBac HT A",
    "host": "insect",
    "category": "baculovirus",
    "size_bp": 4856,
    "promoter": "polyhedrin (PH)",
    "induction": "baculovirus infection",
    "selection": [
      "ampicillin",
      "gentamicin"
    ],
    "copy": "medium",
    "fusion_tag": {
      "n_term": "His6 + TEV protease site (ENLYFQ↓G)",
      "c_term": "none"
    },
    "mcs_enzymes": [
      "BamHI",
      "EcoRI",
      "StuI",
      "SalI",
      "NotI",
      "NcoI",
      "PstI",
      "HindIII",
      "KpnI"
    ],
    "flanking_5p_max": "GGATCC",
    "flanking_3p_max": "AAGCTT",
    "supplier": "Invitrogen / Thermo Fisher",
    "notes": "Bac-to-Bac with N-terminal His6 + TEV site for cleavable affinity purification. Three frame variants (A/B/C)."
  },
  "prsf_duet": {
    "id": "prsf_duet",
    "name": "pRSFDuet-1",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 3829,
    "promoter": "two T7 lac (dual MCS)",
    "induction": "IPTG",
    "selection": [
      "kanamycin"
    ],
    "copy": "high (RSF ori)",
    "fusion_tag": {
      "n_term": "His6 (MCS1)",
      "c_term": "S-tag (MCS2)"
    },
    "mcs_enzymes": [
      "NcoI",
      "BamHI",
      "EcoRI",
      "SacI",
      "NotI",
      "SalI",
      "HindIII",
      "PstI",
      "KpnI",
      "AvrII",
      "XhoI"
    ],
    "flanking_5p_max": "CCATGG",
    "flanking_3p_max": "CTCGAG",
    "notes": "Highest-copy Duet vector. Compatible with pET / pCDF / pACYC Duets for multi-protein co-expression."
  },
  "pet24a": {
    "id": "pet24a",
    "name": "pET-24a(+)",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 5310,
    "promoter": "T7 lac",
    "induction": "IPTG",
    "selection": [
      "kanamycin"
    ],
    "copy": "medium",
    "fusion_tag": {
      "n_term": "T7 tag (optional)",
      "c_term": "LEHHHHHH"
    },
    "mcs_enzymes": [
      "NdeI",
      "NheI",
      "BamHI",
      "EcoRI",
      "SacI",
      "SalI",
      "HindIII",
      "NotI",
      "XhoI"
    ],
    "flanking_5p_max": "AAGGAGATATACATATG",
    "flanking_3p_max": "CTCGAGCACCACCACCACCACCAC",
    "notes": "Kanamycin-resistant version of pET-21a — C-terminal His6 only, no N-tag by default."
  },
  "pet43_1a": {
    "id": "pet43_1a",
    "name": "pET-43.1a (NusA)",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 7287,
    "promoter": "T7 lac",
    "induction": "IPTG",
    "selection": [
      "ampicillin"
    ],
    "copy": "medium",
    "fusion_tag": {
      "n_term": "NusA (~55 kDa) + His6 + thrombin",
      "c_term": "His6 + HSV-tag"
    },
    "mcs_enzymes": [
      "BamHI",
      "EcoRI",
      "SacI",
      "NotI",
      "EagI",
      "PstI",
      "SalI",
      "HindIII",
      "XhoI"
    ],
    "flanking_5p_max": "GGATCC",
    "flanking_3p_max": "CTCGAG",
    "supplier": "Sigma / Novagen",
    "notes": "NusA is the most effective E. coli solubility tag for aggregation-prone proteins (better than MBP/GST in many cases)."
  },
  "ptyb1": {
    "id": "ptyb1",
    "name": "pTYB1 (IMPACT)",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 7477,
    "promoter": "T7",
    "induction": "IPTG",
    "selection": [
      "ampicillin"
    ],
    "copy": "medium",
    "fusion_tag": {
      "n_term": "none",
      "c_term": "intein + chitin binding domain (CBD)"
    },
    "mcs_enzymes": [
      "NdeI",
      "NcoI",
      "NotI",
      "EcoRI",
      "BamHI",
      "EagI",
      "SmaI",
      "SapI"
    ],
    "flanking_5p_max": "CATATG",
    "flanking_3p_max": "TGCTCTTCCAAC",
    "supplier": "NEB (IMPACT system)",
    "notes": "C-terminal intein + CBD. Self-cleaves on chitin column with DTT — releases tag-free native protein with no proteolysis step."
  },
  "pcold_gst": {
    "id": "pcold_gst",
    "name": "pCold GST",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 5616,
    "promoter": "cspA (cold-shock)",
    "induction": "15 °C cold shock + IPTG",
    "selection": [
      "ampicillin"
    ],
    "copy": "medium",
    "fusion_tag": {
      "n_term": "GST + His6 + thrombin",
      "c_term": "none"
    },
    "mcs_enzymes": [
      "NdeI",
      "SacI",
      "KpnI",
      "XhoI",
      "BamHI",
      "EcoRI",
      "HindIII"
    ],
    "flanking_5p_max": "CATATG",
    "flanking_3p_max": "GGATCC",
    "supplier": "Takara Bio",
    "notes": "Cold-shock + GST solubility fusion — useful when both temperature control and GST tag are needed."
  },
  "pbr322": {
    "id": "pbr322",
    "name": "pBR322",
    "host": "e_coli",
    "category": "general_cloning",
    "size_bp": 4361,
    "promoter": "tet, amp (native promoters)",
    "induction": "n/a",
    "selection": [
      "ampicillin",
      "tetracycline"
    ],
    "copy": "medium (15–20 / cell)",
    "mcs_enzymes": [
      "EcoRI",
      "HindIII",
      "BamHI",
      "SalI",
      "PstI",
      "SphI",
      "ClaI",
      "NruI",
      "AvaI",
      "PvuII",
      "ScaI",
      "XmaIII"
    ],
    "flanking_5p_max": "GAATTC",
    "flanking_3p_max": "AAGCTT",
    "notes": "Classic foundational cloning vector. Lower copy than pUC19 — better for unstable inserts. Two antibiotic markers."
  },
  "pentr_d_topo": {
    "id": "pentr_d_topo",
    "name": "pENTR/D-TOPO (Gateway)",
    "host": "e_coli",
    "category": "gateway",
    "size_bp": 2580,
    "promoter": "none (entry clone)",
    "induction": "n/a",
    "selection": [
      "kanamycin"
    ],
    "copy": "high",
    "mcs_enzymes": [
      "(directional TOPO — CACC overhang)"
    ],
    "flanking_5p_max": "CACC",
    "flanking_3p_max": "",
    "supplier": "Invitrogen / Thermo Fisher",
    "notes": "Gateway entry vector. Clone via directional TOPO (5′ CACC overhang), then LR-recombine into any pDEST destination vector."
  },
  "pdonr221": {
    "id": "pdonr221",
    "name": "pDONR221 (Gateway)",
    "host": "e_coli",
    "category": "gateway",
    "size_bp": 4762,
    "promoter": "none (donor)",
    "induction": "n/a",
    "selection": [
      "kanamycin",
      "ccdB counterselection"
    ],
    "copy": "high",
    "mcs_enzymes": [
      "(attP1 / attP2 — BP clonase)"
    ],
    "flanking_5p_max": "GGGGACAAGTTTGTACAAAAAAGCAGGCT",
    "flanking_3p_max": "GGGGACCACTTTGTACAAGAAAGCTGGGT",
    "supplier": "Invitrogen",
    "notes": "Gateway donor vector. BP-recombine attB-flanked PCR product to make entry clone; then LR into destination."
  },
  "pbabe_puro": {
    "id": "pbabe_puro",
    "name": "pBABE-puro",
    "host": "mammalian",
    "category": "retroviral",
    "size_bp": 5169,
    "promoter": "5′ MoMuLV LTR (transgene) + SV40 (Puro)",
    "induction": "constitutive",
    "selection": [
      "ampicillin (E. coli)",
      "puromycin (mammalian)"
    ],
    "copy": "integrated",
    "mcs_enzymes": [
      "BamHI",
      "SnaBI",
      "SalI",
      "EcoRI",
      "NheI"
    ],
    "flanking_5p_max": "GGATCC",
    "flanking_3p_max": "GTCGAC",
    "addgene": "1764",
    "notes": "Classic Moloney MLV retroviral transfer vector. Co-transfect with packaging plasmids in Phoenix or 293T cells."
  },
  "plvx_ires_puro": {
    "id": "plvx_ires_puro",
    "name": "pLVX-IRES-Puro",
    "host": "mammalian",
    "category": "lentiviral",
    "size_bp": 8155,
    "promoter": "CMV (transgene + IRES + Puro)",
    "induction": "constitutive",
    "selection": [
      "ampicillin (E. coli)",
      "puromycin (mammalian)"
    ],
    "copy": "integrated",
    "mcs_enzymes": [
      "EcoRI",
      "BamHI",
      "XhoI",
      "NotI",
      "MluI",
      "XbaI",
      "SpeI"
    ],
    "flanking_5p_max": "GAATTC",
    "flanking_3p_max": "GCGGCCGC",
    "supplier": "Clontech / Takara",
    "notes": "3rd-gen lentivirus + IRES bicistronic Puro selection. Transgene and Puro from one transcript."
  },
  "pmscv_puro": {
    "id": "pmscv_puro",
    "name": "pMSCV-puro",
    "host": "mammalian",
    "category": "retroviral",
    "size_bp": 6347,
    "promoter": "MSCV 5′ LTR + PGK (Puro)",
    "induction": "constitutive",
    "selection": [
      "ampicillin",
      "puromycin"
    ],
    "copy": "integrated",
    "mcs_enzymes": [
      "BglII",
      "HpaI",
      "XhoI",
      "EcoRI"
    ],
    "flanking_5p_max": "AGATCT",
    "flanking_3p_max": "GAATTC",
    "supplier": "Clontech",
    "notes": "Murine stem cell virus (MSCV) LTR works well in hematopoietic / stem cells where MoMuLV LTR is silenced."
  },
  "pci_neo": {
    "id": "pci_neo",
    "name": "pCI-neo",
    "host": "mammalian",
    "category": "mammalian_expression",
    "size_bp": 5472,
    "promoter": "CMV (+ chimeric intron)",
    "induction": "constitutive",
    "selection": [
      "ampicillin",
      "G418/neomycin"
    ],
    "copy": "episomal",
    "mcs_enzymes": [
      "NheI",
      "EcoRI",
      "SacI",
      "KpnI",
      "MluI",
      "XhoI",
      "NotI",
      "XbaI",
      "BamHI",
      "SalI"
    ],
    "flanking_5p_max": "GCTAGC",
    "flanking_3p_max": "TCTAGA",
    "supplier": "Promega",
    "notes": "CMV + chimeric intron boosts expression ~2-fold over pcDNA3.1 in transient transfections."
  },
  "pcag_gfp": {
    "id": "pcag_gfp",
    "name": "pCAG-GFP",
    "host": "mammalian",
    "category": "mammalian_expression",
    "size_bp": 6230,
    "promoter": "CAG (CMV enhancer + chicken β-actin + rabbit β-globin intron)",
    "induction": "constitutive",
    "selection": [
      "ampicillin"
    ],
    "copy": "episomal",
    "fusion_tag": {
      "n_term": "none",
      "c_term": "EGFP"
    },
    "mcs_enzymes": [
      "EcoRI",
      "AgeI",
      "NheI"
    ],
    "flanking_5p_max": "GAATTC",
    "flanking_3p_max": "ACCGGT",
    "addgene": "11150",
    "notes": "CAG promoter gives very strong constitutive expression across many cell types — used in vivo (mouse) and in iPSC."
  },
  "pad_cmv_v5_dest": {
    "id": "pad_cmv_v5_dest",
    "name": "pAd/CMV/V5-DEST",
    "host": "mammalian",
    "category": "adenoviral",
    "size_bp": 36702,
    "promoter": "CMV",
    "induction": "constitutive",
    "selection": [
      "ampicillin (E. coli)"
    ],
    "copy": "packaged into Ad5 particles",
    "fusion_tag": {
      "n_term": "none (optional)",
      "c_term": "V5 + His6 (optional)"
    },
    "mcs_enzymes": [
      "(attR1 / attR2 — Gateway LR)"
    ],
    "flanking_5p_max": "CAAGTTTGTACAAAAAAGCTGAAC",
    "flanking_3p_max": "CAACTTTGTATAATAAAGTTG",
    "supplier": "Invitrogen",
    "notes": "Adenoviral expression via Gateway LR. Linearize with PacI, transfect 293A cells to produce Ad5 virus."
  },
  "pcdh1_mcs1": {
    "id": "pcdh1_mcs1",
    "name": "pCDH1-MCS1-EF1-copGFP",
    "host": "mammalian",
    "category": "lentiviral",
    "size_bp": 7559,
    "promoter": "CMV (transgene) + EF1α (copGFP)",
    "induction": "constitutive",
    "selection": [
      "ampicillin (E. coli)",
      "copGFP flow-sort (mammalian)"
    ],
    "copy": "integrated",
    "mcs_enzymes": [
      "EcoRI",
      "BamHI",
      "NheI",
      "NotI",
      "XbaI",
      "EcoRV",
      "SwaI",
      "ClaI",
      "SalI"
    ],
    "flanking_5p_max": "GAATTC",
    "flanking_3p_max": "GCGGCCGC",
    "supplier": "System Biosciences (SBI)",
    "notes": "Lentiviral transfer with copGFP marker — use FACS instead of antibiotic selection for transduced cells."
  },
  "px458": {
    "id": "px458",
    "name": "pSpCas9(BB)-2A-GFP (PX458)",
    "host": "mammalian",
    "category": "mammalian_expression",
    "size_bp": 9288,
    "promoter": "CBh (Cas9) + U6 (sgRNA)",
    "induction": "constitutive",
    "selection": [
      "ampicillin (E. coli)",
      "GFP FACS (mammalian)"
    ],
    "copy": "episomal",
    "fusion_tag": {
      "n_term": "none",
      "c_term": "2A-EGFP (Cas9 only)"
    },
    "mcs_enzymes": [
      "BbsI"
    ],
    "flanking_5p_max": "CACCG",
    "flanking_3p_max": "GTTT",
    "addgene": "48138",
    "notes": "Cas9 + sgRNA + 2A-GFP. Sort transfected cells by GFP+. Zhang lab."
  },
  "pmaltose_e": {
    "id": "pmaltose_e",
    "name": "pMAL-p5X (periplasmic)",
    "host": "e_coli",
    "category": "bacterial_expression",
    "size_bp": 5752,
    "promoter": "tac",
    "induction": "IPTG",
    "selection": [
      "ampicillin"
    ],
    "copy": "medium",
    "fusion_tag": {
      "n_term": "malE signal + MBP + Factor Xa",
      "c_term": "none"
    },
    "mcs_enzymes": [
      "NdeI",
      "EcoRI",
      "BamHI",
      "XbaI",
      "SalI",
      "PstI",
      "HindIII"
    ],
    "flanking_5p_max": "GAAGGTAGGTCATATG",
    "flanking_3p_max": "AAGCTT",
    "supplier": "NEB",
    "notes": "Periplasmic variant of pMAL — malE signal exports fusion to periplasm. Cytoplasmic MBP folding bypassed."
  }
}