LOCUS       sequence-235452-        3534 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 3534)
  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 3534)
  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: TRAV12-2
FEATURES             Location/Qualifiers
     source          1..3534
                     /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(1444..1460)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1444..1460)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1457..1479)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1468..1484
                     /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(1492..1498,1499..1516,1517..1522))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1537..1558
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1675..1692)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1846..2434)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1926..1945)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2612..3421)
                     /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     3335..3354
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3413..3432
                     /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 ggcccatgaa atccttgaga gttttactag
     1081 tgatcctgtg gcttcagttg agctgggttt ggagccaaca gaaggaggtg gagcagaatt
     1141 ctggacccct cagtgttcca gagggagcca ttgcctctct caactgcact tacagtgacc
     1201 gaggttccca gtccttcttc tggtacagac aatattctgg gaaaagccct gagttgataa
     1261 tgttcatata ctccaatggt gacaaagaag atggaaggtt tacagcacag ctcaataaag
     1321 ccagccagta tgtttctctg ctcatcagag actcccagcc cagtgattca gccacctact
     1381 gagaccatcg gatcccgggc ccgtcgactg cagaggcctg catgcaagct tggtgtaatc
     1441 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     1501 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     1561 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     1621 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     1681 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     1741 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     1801 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1861 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1921 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1981 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2041 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2101 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2161 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2221 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2281 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2341 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2401 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2461 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2521 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa gcccaatctg
     2581 aataatgtta caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa
     2641 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     2701 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
     2761 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     2821 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat
     2881 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     2941 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
     3001 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
     3061 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgtttttc
     3121 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     3181 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     3241 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     3301 agcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     3361 aatcagcatc catgttggaa tttaatcgcg gcctcgacgt ttcccgttga atatggctca
     3421 taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat
     3481 ttttatcttg tgcaatgtaa catcagagat tttgagacac gggccagagc tgca
//
