LOCUS       sequence-244186-        3531 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 3531)
  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 3531)
  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: TRAV41
FEATURES             Location/Qualifiers
     source          1..3531
                     /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(1256..1272)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1256..1272)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1269..1291)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1280..1296
                     /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(1304..1310,1311..1328,1329..1334))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1349..1370
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1487..1504)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1658..2246)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1738..1757)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2424..3233)
                     /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     3147..3166
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3225..3244
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3375..3397)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3497..3516
                     /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 atggtgaaga tccggcaatt tttgttggct
      901 attttgtggc ttcagctaag ctgtgtaagt gccgccaaaa atgaagtgga gcagagtcct
      961 cagaacctga ctgcccagga aggagaattt atcacaatca actgcagtta ctcggtagga
     1021 ataagtgcct tacactggct gcaacagcat ccaggaggag gcattgtttc cttgtttatg
     1081 ctgagctcag ggaagaagaa gcatggaaga ttaattgcca caataaacat acaggaaaag
     1141 cacagctccc tgcacatcac agcctcccat cccagagact ctgccgtcta ctgagaccat
     1201 cggatcccgg gcccgtcgac tgcagaggcc tgcatgcaag cttggtgtaa tcatggtcat
     1261 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     1321 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     1381 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     1441 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     1501 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     1561 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     1621 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     1681 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     1741 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     1801 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     1861 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1921 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1981 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     2041 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     2101 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     2161 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     2221 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2281 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     2341 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aagcccaatc tgaataatgt
     2401 tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg aaactgcaat
     2461 ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga
     2521 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg
     2581 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt
     2641 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt atgcatttct
     2701 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc
     2761 aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa
     2821 ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca
     2881 atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt tccggggatc
     2941 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga
     3001 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg
     3061 ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata caagcgatag
     3121 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca
     3181 tccatgttgg aatttaatcg cggcctcgac gtttcccgtt gaatatggct cataacaccc
     3241 cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat atttttatct
     3301 tgtgcaatgt aacatcagag attttgagac acgggccaga gctgcatcgc gcgtttcggt
     3361 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     3421 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     3481 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca t
//
