LOCUS       sequence-244184-        3540 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 3540)
  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 3540)
  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: TRAV38-2
FEATURES             Location/Qualifiers
     source          1..3540
                     /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(1265..1281)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1265..1281)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1278..1300)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1289..1305
                     /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(1313..1319,1320..1337,1338..1343))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1358..1379
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1496..1513)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1667..2255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1747..1766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2433..3242)
                     /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     3156..3175
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3234..3253
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3384..3406)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3506..3525
                     /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 atggcatgcc ctggcttcct gtgggcactt
      901 gtgatctcca cctgtcttga atttagcatg gctcagacag tcactcagtc tcaaccagag
      961 atgtctgtgc aggaggcaga gacagtgacc ctgagctgca catatgacac cagtgagagt
     1021 gattattatt tattctggta caagcagcct cccagcaggc agatgattct cgttattcgc
     1081 caagaagctt ataagcaaca gaatgcaaca gagaatcgtt tctctgtgaa cttccagaaa
     1141 gcagccaaat ccttcagtct caagatctca gactcacagc tgggggatgc cgcgatgtat
     1201 tgagaccatc ggatcccggg cccgtcgact gcagaggcct gcatgcaagc ttggtgtaat
     1261 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     1321 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     1381 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     1441 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     1501 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1561 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1621 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1681 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1741 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1801 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     1861 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     1921 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     1981 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2041 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2101 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2161 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2221 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2281 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2341 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca agcccaatct
     2401 gaataatgtt acaaccaatt aaccaattct gattagaaaa actcatcgag catcaaatga
     2461 aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt
     2521 aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct
     2581 gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg
     2641 ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta
     2701 tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc
     2761 gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg
     2821 ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc
     2881 gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttt
     2941 ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg
     3001 gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca
     3061 ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac
     3121 aagcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat
     3181 aaatcagcat ccatgttgga atttaatcgc ggcctcgacg tttcccgttg aatatggctc
     3241 ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata
     3301 tttttatctt gtgcaatgta acatcagaga ttttgagaca cgggccagag ctgcatcgcg
     3361 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     3421 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     3481 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
//
