LOCUS       sequence-475800-        3198 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes TRAV4 allele, P2A, and TRBC to generate TCRs via cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3198)
  AUTHORS   Gaglione SA, Mukkamala RS, Krishna C, Smith BE, Wadsworth II MH, 
            Jelinsky SA, Perez CR, Schmidt-Hong L, Katz EL, Gellatly KJ, Ali LR,
            Shen J, Holec PV, Zhao QH, Chan AO , Xu EJK, Kravarik KM, Guzova JA,
            Dobson CS, Singh H, Garber M, Dougan M, Dougan SK, Harris JE, 
            Winkler A, Birnbaum ME
  TITLE     Scalable TCR synthesis and screening enables antigen reactivity 
            mapping in vitiligo
  JOURNAL   bioRxiv 2025.04.24.649726
REFERENCE   2  (bases 1 to 3198)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2025.04.24.649726"
COMMENT     Sequence Label: TRAV4
FEATURES             Location/Qualifiers
     source          1..3198
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(57..73)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(57..73)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(70..92)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(245..833)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(325..344)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1014..1823)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1737..1756
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1815..1834
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(1965..1987)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2172..2194
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2186..2203
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2187..2203
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2811..2867
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
ORIGIN
        1 aggctaggtg gaggctcagt gatgataagt ctgcgatggt ggatgcatgt gtcatggtca
       61 tagctgtttc ctgtgtgaaa ttgttatccg ctcagagggc acaatcctat tccgcgctat
      121 ccgacaatct ccaagacatt aggtggagtt cagttcggcg tatggcatat gtcgctggaa
      181 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      241 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      301 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
      361 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
      421 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
      481 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
      541 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
      601 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
      661 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
      721 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
      781 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagaag
      841 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
      901 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
      961 gttttaaatc aagcccaatc tgaataatgt tacaaccaat taaccaattc tgattagaaa
     1021 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     1081 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     1141 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     1201 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     1261 ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta
     1321 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     1381 gcaagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     1441 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     1501 aatacctgga atgctgtttt tccggggatc gcagtggtga gtaaccatgc atcatcagga
     1561 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     1621 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     1681 ggcgcatcgg gcttcccata caagcgatag attgtcgcac ctgattgccc gacattatcg
     1741 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgac
     1801 gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta agcagacagt
     1861 tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag attttgagac
     1921 acgggccaga gctgcatcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     1981 ctcccgggac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     2041 gcgcgtcagc gggtgttggc tgttgtttgt cgcattatac gagcgtccag gttgggatac
     2101 ctgaaacaaa acccatcgta cggccaagga agtctccaat aactgtgatc caccacaagc
     2161 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtcatgcat aatccgcacg
     2221 catctggaat aaggaagtgc cattccgcct gacctctcga gtaatagaga cccgtctcag
     2281 tattcccacc cgaggtcgct gtgtttgagc catcagaagc agagatctcc cacacccaaa
     2341 aggccacact ggtgtgcctg gccacaggct tctaccccga ccacgtggag ctgagctggt
     2401 gggtgaatgg gaaggaggtg cacagtgggg tcagcacaga cccgcagccc ctcaaggagc
     2461 agcccgccct caatgactcc agatactgcc tgagcagccg cctgagagtc tcggccacct
     2521 tctggcagaa cccccgcaac cacttccgct gtcaagtcca gttctacggg ctctcggaga
     2581 atgacgagtg gacccaggat agggccaaac ctgtcaccca gatcgtcagc gccgaggcct
     2641 ggggtagagc agactgtggc ttcacctccg agtcttacca gcaaggggtc ctgtctgcca
     2701 ccatcctcta tgagatcttg ctagggaagg ccaccttgta tgccgtgctg gtcagtgccc
     2761 tcgtgctgat ggccatggtc aagagaaagg attccagagg cggaagtgga gcaaccaatt
     2821 tctccttgct gaagcaggcc ggtgacgttg aggaaaaccc aggaccaatg aggcaagtgg
     2881 cgagagtgat cgtgttcctg accctgagta ctttgagcct tgctaagacc acccagccca
     2941 tctccatgga ctcatatgaa ggacaagaag tgaacataac ctgtagccac aacaacattg
     3001 ctacaaatga ttatatcacg tggtaccaac agtttcccag ccaaggacca cgatttatta
     3061 ttcaaggata caagacaaaa gttacaaacg aagtggcctc cctgtttatc cctgccgaca
     3121 gaaagtccag cactctgagc ctgccccggg tttccctgag cgacactgca gtgggagacg
     3181 cggtctcaaa acctcgag
//
