LOCUS       sequence-235486-        3424 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 3424)
  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 3424)
  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: TRBV5-6
FEATURES             Location/Qualifiers
     source          1..3424
                     /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"
     regulatory      447..456
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(765..776)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1334..1350)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1334..1350)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1347..1369)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1358..1374
                     /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(1382..1388,1389..1406,1407..1412))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1427..1448
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1565..1582)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1736..2324)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1816..1835)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2502..3311)
                     /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     3225..3244
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3303..3322
                     /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 tggggatcca ccggtcgcca ccatgggccc cgggctcctc tgctgggcac
      481 tgctttgtct cctgggagca ggcttagtgg acgctggagt cacccaaagt cccacacacc
      541 tgatcaaaac gagaggacag caagtgactc tgagatgctc tcctaagtct gggcatgaca
      601 ctgtgtcctg gtaccaacag gccctgggtc aggggcccca gtttatcttt cagtattatg
      661 aggaggaaga gagacagaga ggcaacttcc ctgatcgatt ctcaggtcac cagttcccta
      721 actatagctc tgagctgaat gtgaacgcct tgttgctggg ggactcggcc ctctatgaga
      781 cctaaggtgg tggtggttct ggttctggtt ctggtggtgg ttctggtggt ggtctctgaa
      841 ccctgaccct gccgtgtacc agctgagaga ctctaaatcc agtgacaagt ctgtctgcct
      901 attcaccgat tttgattctc aaacaaatgt gtcacaaagt aaggattctg atgtgtatat
      961 cacagacaaa actgtgctag acatgaggtc tatggacttc aagagcaaca gtgctgtggc
     1021 ctggagcaac aaatctgact ttgcatgtgc aaacgccttc aacaacagca ttattccaga
     1081 agacaccttc ttccccagcc cagaaagttc ctgtgatgtc aagctggtcg agaaaagctt
     1141 tgaaacagat acgaacctaa actttcaaaa cctgtcagtg attgggttcc gaatcctcct
     1201 cctgaaagtg gccgggttta atctgctcat gacgctgcgg ctgtggtcca gctaaagcgg
     1261 ccgcgtcgac aatcaaatcg gatcccgggc ccgtcgactg cagaggcctg catgcaagct
     1321 tggtgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     1381 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     1441 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     1501 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     1561 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     1621 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     1681 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1741 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1801 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1861 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1921 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     1981 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2041 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2101 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2161 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2221 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2281 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2341 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2401 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2461 gcccaatctg aataatgtta caaccaatta accaattctg attagaaaaa ctcatcgagc
     2521 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     2581 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     2641 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     2701 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     2761 aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     2821 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     2881 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     2941 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     3001 gctgtttttc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     3061 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     3121 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     3181 ttcccataca agcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     3241 tacccatata aatcagcatc catgttggaa tttaatcgcg gcctcgacgt ttcccgttga
     3301 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     3361 gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac gggccagagc
     3421 tgca
//
