LOCUS       sequence-244185-        3525 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  construction of TCR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3525)
  AUTHORS   Hu Z, Anandappa AJ, Sun J, Kim J, Leet DE, Bozym DJ, Chen C, 
            Williams L, Shukla SA, Zhang W, Tabbaa D, Steelman S, Olive O, Livak
            KJ, Kishi H, Muraguchi A, Guleria I, Stevens J, Lane WJ, Burkhardt 
            UE, Fritsch EF, Neuberg D, Ott PA, Keskin DB, Hacohen N, Wu CJ
  TITLE     A cloning and expression system to probe T-cell receptor specificity
            and assess functional avidity to neoantigens.
  JOURNAL   Blood. 2018 Nov 1;132(18):1911-1921. doi: 
            10.1182/blood-2018-04-843763. Epub 2018 Aug 27.
  PUBMED    30150207
REFERENCE   2  (bases 1 to 3525)
  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"; doi: 
            "10.1182/blood-2018-04-843763"; journalName: "Blood"; date: 
            "2018-11-1- 1"; volume: "132"; issue: "18"; pages: "1911-1921"
COMMENT     Sequence Label: TRAV39
FEATURES             Location/Qualifiers
     source          1..3525
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     179..201
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             805..870
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     primer_bind     complement(1250..1266)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1250..1266)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1263..1285)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1274..1290
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1298..1304,1305..1322,1323..1328))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1343..1364
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1481..1498)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1652..2240)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1732..1751)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2418..3227)
                     /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     3141..3160
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3219..3238
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3369..3391)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3491..3510
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
       61 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
      121 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
      181 cagtcacgac gttgtaaaac gacggccaga gaattcgagc tcggtacctc gcgaatacat
      241 ctagatggtc tctacctgaa caaggtgttc ccacccgagg tcgctgtgtt tgagccatca
      301 gaagcagaga tctcccacac ccaaaaggcc acactggtgt gcctggccac aggcttcttc
      361 cccgaccacg tggagctgag ctggtgggtg aatgggaagg aggtgcacag tggggtcagc
      421 acggacccgc agcccctcaa ggagcagccc gccctcaatg actccagata ctgcctgagc
      481 agccgcctga gggtttcggc caccttctgg cagaaccccc gcaaccactt ccgctgtcaa
      541 gtccagttct acgggctctc ggagaatgac gagtggaccc aggatagggc caaacccgtc
      601 acccagatcg tcagcgccga ggcctggggt agagcagact gtggctttac ctcggtgtcc
      661 taccagcaag gggtcctgtc tgccaccatc ctctatgaga tcctgctagg gaaggccacc
      721 ctgtatgctg tgctggtcag cgcccttgtg ttgatggcca tggtcaagag aaaggatttc
      781 aggaggagga ggagcggcag tggagtgaaa cagactttga attttgacct tctcaagttg
      841 gcgggagacg tggagtccaa cccagggccc atgaagaagc tactagcaat gattctgtgg
      901 cttcaactag accgcttaag tggagagctg aaagtggaac aaaaccctct gttcctgagc
      961 atgcaggagg gaaaaaacta taccatctac tgcaattatt caaccacttc agacagactg
     1021 tattggtaca ggcaggatcc tgggaaaagt ctggaatctc tgtttgtgtt gctatcaaat
     1081 ggagcagtga agcaggaggg acgattaatg gcctcacttg ataccaaagc ccgtctcagc
     1141 accctccaca tcacagctgc cgtgcatgac ctctctgcca cctactgaga ccatcggatc
     1201 ccgggcccgt cgactgcaga ggcctgcatg caagcttggt gtaatcatgg tcatagctgt
     1261 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     1321 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     1381 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     1441 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     1501 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     1561 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     1621 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1681 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1741 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1801 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1861 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1921 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1981 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2041 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2101 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2161 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2221 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2281 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2341 agatcctttt aaattaaaaa tgaagtttta aatcaagccc aatctgaata atgttacaac
     2401 caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc
     2461 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     2521 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     2581 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     2641 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag
     2701 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     2761 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa
     2821 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     2881 tcacctgaat caggatattc ttctaatacc tggaatgctg tttttccggg gatcgcagtg
     2941 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     3001 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     3061 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaagcg atagattgtc
     3121 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     3181 ttggaattta atcgcggcct cgacgtttcc cgttgaatat ggctcataac accccttgta
     3241 ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca
     3301 atgtaacatc agagattttg agacacgggc cagagctgca tcgcgcgttt cggtgatgac
     3361 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     3421 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     3481 cttaactatg cggcatcaga gcagattgta ctgagagtgc accat
//
