LOCUS       sequence-235460-        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: TRAV20
FEATURES             Location/Qualifiers
     source          1..3531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             990..1055
                     /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(1441..1457)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1441..1457)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1454..1476)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1465..1481
                     /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(1489..1495,1496..1513,1514..1519))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1534..1555
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1672..1689)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1843..2431)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1923..1942)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2609..3418)
                     /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     3332..3351
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3410..3429
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagagaatt cgagctcggt acctcgcgaa
      421 tacatctaga tggtctctac ctgaacaagg tgttcccacc cgaggtcgct gtgtttgagc
      481 catcagaagc agagatctcc cacacccaaa aggccacact ggtgtgcctg gccacaggct
      541 tcttccccga ccacgtggag ctgagctggt gggtgaatgg gaaggaggtg cacagtgggg
      601 tcagcacgga cccgcagccc ctcaaggagc agcccgccct caatgactcc agatactgcc
      661 tgagcagccg cctgagggtt tcggccacct tctggcagaa cccccgcaac cacttccgct
      721 gtcaagtcca gttctacggg ctctcggaga atgacgagtg gacccaggat agggccaaac
      781 ccgtcaccca gatcgtcagc gccgaggcct ggggtagagc agactgtggc tttacctcgg
      841 tgtcctacca gcaaggggtc ctgtctgcca ccatcctcta tgagatcctg ctagggaagg
      901 ccaccctgta tgctgtgctg gtcagcgccc ttgtgttgat ggccatggtc aagagaaagg
      961 atttcaggag gaggaggagc ggcagtggag tgaaacagac tttgaatttt gaccttctca
     1021 agttggcggg agacgtggag tccaacccag ggcccatgga gaaaatgttg gagtgtgcat
     1081 tcatagtctt gtggcttcag cttggctggt tgagtggaga agaccaggtg acgcagagtc
     1141 ccgaggccct gagactccag gagggagaga gtagcagtct taactgcagt tacacagtca
     1201 gcggtttaag agggctgttc tggtataggc aagatcctgg gaaaggccct gaattcctct
     1261 tcaccctgta ttcagctggg gaagaaaagg agaaagaaag gctaaaagcc acattaacaa
     1321 agaaggaaag ctttctgcac atcacagccc ctaaacctga agactcagcc acttattgag
     1381 accatcggat cccgggcccg tcgactgcag aggcctgcat gcaagcttgg tgtaatcatg
     1441 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     1501 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     1561 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     1621 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     1681 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1741 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1801 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1861 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1921 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1981 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2041 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2101 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2161 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2221 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2281 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2341 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2401 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2461 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2521 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaagcc caatctgaat
     2581 aatgttacaa ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact
     2641 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
     2701 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     2761 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     2821 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca
     2881 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     2941 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt
     3001 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     3061 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gtttttccgg
     3121 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     3181 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     3241 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaagc
     3301 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     3361 cagcatccat gttggaattt aatcgcggcc tcgacgtttc ccgttgaata tggctcataa
     3421 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt
     3481 tatcttgtgc aatgtaacat cagagatttt gagacacggg ccagagctgc a
//
