LOCUS       sequence-475857-        2918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes TRBV27 allele and TRAC to generate TCRs via cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2918)
  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 2918)
  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: TRBV27
FEATURES             Location/Qualifiers
     source          1..2918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(436..455)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(437..455)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(566..1225)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1140..1159
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1226..1328)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      1429..2017
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1918..1937
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2171..2188
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        2447..2465
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2447..2464
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      2513..2522
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ctgagagact ctaaatccag tgacaagtct gtctgcctat tcaccgattt tgattctcaa
       61 acaaatgtgt cacaaagtaa ggattctgac gtctatatca cagacaaaac tgtgctagac
      121 atgaggtcta tggacttcaa gagcaacagt gctgtggcct ggagcaacaa atctgacttt
      181 gcatgtgcaa acgccttcaa caacagcatt attccagaag ataccttctt ccccagccca
      241 gaaagttcct gtgatgtcaa gctggtcgag aaatcttttg aaacagatac gaacctaaac
      301 tttcaaaacc tgtcagtgat tgggttccga atcctcctcc tgaaagtggc cgggtttaat
      361 ctgctcatga cgctgcggct gtggtccagc tgaggaattc atcagaagag cctcgaggcg
      421 gccgcatgtg agtctcccta tagtgagtcg tattaatttc gcgggcggaa cccctatttg
      481 tttatttttc taaatacatt caaatatgta tccgctcatg agtagcacca ggcgtttaag
      541 ggcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg cagtactgtt
      601 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
      661 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg
      721 gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg
      781 gattggctga gacaaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt
      841 caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt
      901 attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt
      961 gaacactatc ccatatcacc agctcaccgt ctttcattgc catacgaaat tccggatgag
     1021 cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct
     1081 ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag
     1141 caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg
     1201 tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact
     1261 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     1321 gccgatcaaa gtctcatttt cgccaaaagt tgtcatgacc aaaatccctt aacgtgagtt
     1381 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     1441 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     1501 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     1561 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     1621 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     1681 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     1741 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     1801 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     1861 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     1921 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     1981 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaatg cggccttttt
     2041 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2101 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     2161 gaccgagcgc agcgagtcag tgagcgagga agccgaaaaa tcaataatca gacaacaaga
     2221 tgtgcgaact cgatatttta cacgactctc tttaccaatt ctgccccgaa ttacacttaa
     2281 aacgactcaa cagcttaacg ttggcttgcc acgcattact tgactgtaaa actctcactc
     2341 ttaccgaact tggccgtaac ctgccaacca aagcgagaac aaaacataac atcaaacgaa
     2401 tcgaccgatt gttaggtaat cgtcacttgc aggaaggttt aaacgcattt aggtgacact
     2461 atagaagtgt gtatcgctcg aggctcttca gcatctagag ccaccggcat gagccaccat
     2521 gggcccccag ctccttggct atgtggtcct ttgccttcta ggagcaggcc ccctggaagc
     2581 ccaagtgacc cagaacccaa gatacctcat cacagtgact ggaaagaagt taacagtgac
     2641 ttgttctcag aatatgaacc atgagtatat gtcctggtat cgacaagacc cagggctggg
     2701 cttaaggcag atctactatt caatgaatgt tgaggtgact gataagggag atgttcctga
     2761 agggtacaaa gtctctcgaa aagagaagag gaatttcccc ctgatcctgg agtcgcccag
     2821 ccccaaccag acctcgctag tcttcgtcac gagacgtaat ggagaccatg cgatatccgt
     2881 aggtctcgaa accgtctcgg ggtgaagaca tctatcag
//
