LOCUS       sequence-235462-        3525 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 3525)
  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 3525)
  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: TRAV22
FEATURES             Location/Qualifiers
     source          1..3525
                     /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(1435..1451)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1435..1451)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1448..1470)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1459..1475
                     /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(1483..1489,1490..1507,1508..1513))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1528..1549
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1666..1683)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1837..2425)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1917..1936)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2603..3412)
                     /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     3326..3345
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3404..3423
                     /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 gaggatattg ggagctctgc
     1081 tggggctctt gagtgcccag gtttgctgtg tgagaggaat acaagtggag cagagtcctc
     1141 cagacctgat tctccaggag ggagccaatt ccacgctgcg gtgcaatttt tctgactctg
     1201 tgaacaattt gcagtggttt catcaaaacc cttggggaca gctcatcaac ctgttttaca
     1261 ttccctcagg gacaaaacag aatggaagat taagcgccac gactgtcgct acggaacgct
     1321 acagcttatt gtacatttcc tcttcccaga ccacagactc aggcgtttat tgagaccatc
     1381 ggatcccggg cccgtcgact gcagaggcct gcatgcaagc ttggtgtaat catggtcata
     1441 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     1501 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     1561 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     1621 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     1681 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1741 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1801 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1861 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1921 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1981 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2041 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2101 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2161 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2221 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2281 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2341 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2401 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2461 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2521 cacctagatc cttttaaatt aaaaatgaag ttttaaatca agcccaatct gaataatgtt
     2581 acaaccaatt aaccaattct gattagaaaa actcatcgag catcaaatga aactgcaatt
     2641 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     2701 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     2761 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     2821 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
     2881 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     2941 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     3001 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     3061 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttt ccggggatcg
     3121 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     3181 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     3241 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aagcgataga
     3301 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     3361 ccatgttgga atttaatcgc ggcctcgacg tttcccgttg aatatggctc ataacacccc
     3421 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt
     3481 gtgcaatgta acatcagaga ttttgagaca cgggccagag ctgca
//
