LOCUS       sequence-235469-        3552 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 3552)
  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 3552)
  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: TRAV29
FEATURES             Location/Qualifiers
     source          1..3552
                     /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(1462..1478)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1462..1478)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1475..1497)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1486..1502
                     /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(1510..1516,1517..1534,1535..1540))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1555..1576
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1693..1710)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1864..2452)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1944..1963)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2630..3439)
                     /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     3353..3372
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3431..3450
                     /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 ggcccatggc catgctcctg ggggcatcag
     1081 tgctgattct gtggcttcag ccagactggg taaacagtca acagaagaat gatgaccagc
     1141 aagttaagca aaattcacca tccctgagcg tccaggaagg aagaatttct attctgaact
     1201 gtgactatac taacagcatg tttgattatt tcctatggta caaaaaatac cctgctgaag
     1261 gtcctacatt cctgatatct ataagttcca ttaaggataa aaatgaagat ggaagattca
     1321 ctgtcttctt aaacaaaagt gccaagcacc tctctctgca cattgtgccc tcccagcctg
     1381 gagactctgc agtgtactga gaccatcgga tcccgggccc gtcgactgca gaggcctgca
     1441 tgcaagcttg gtgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     1501 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     1561 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     1621 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1681 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1741 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1801 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1861 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1921 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1981 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2041 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2101 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2161 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2221 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2281 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2341 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2401 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2461 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2521 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2581 taaatcaagc ccaatctgaa taatgttaca accaattaac caattctgat tagaaaaact
     2641 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     2701 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     2761 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     2821 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     2881 agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     2941 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     3001 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     3061 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     3121 cctggaatgc tgtttttccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     3181 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     3241 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     3301 catcgggctt cccatacaag cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     3361 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ctcgacgttt
     3421 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     3481 ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacgg
     3541 gccagagctg ca
//
