LOCUS       sequence-245031-        3448 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 3448)
  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 3448)
  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: TRBV13
FEATURES             Location/Qualifiers
     source          1..3448
                     /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"
     primer_bind     complement(1358..1374)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1358..1374)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1371..1393)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1382..1398
                     /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(1406..1412,1413..1430,1431..1436))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1451..1472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1589..1606)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1760..2348)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1840..1859)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2526..3335)
                     /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     3249..3268
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3327..3346
                     /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 ccatgcttag tcctgacctg cctgactctg
      481 cctggaacac caggctcctc tgccgtgtca tgctttgtct cctgggagca ggttcagtgg
      541 ctgctggagt catccagtcc ccaagacatc tgatcaaaga aaagagggaa acagccactc
      601 tgaaatgcta tcctatccct agacacgaca ctgtctactg gtaccagcag ggtccaggtc
      661 aggaccccca gttcctcatt tcgttttatg aaaagatgca gagcgataaa ggaagcatcc
      721 ctgatcgatt ctcagctcaa cagttcagtg actatcattc tgaactgaac atgagctcct
      781 tggagctggg ggactcagcc ctgtacgaga cctaaggtgg tggtggttct ggttctggtt
      841 ctggtggtgg ttctggtggt ggtctcttga ccctgccgtg taccagctga gagactctaa
      901 atccagtgac aagtctgtct gcctattcac cgattttgat tctcaaacaa atgtgtcaca
      961 aagtaaggat tctgatgtgt atatcacaga caaaactgtg ctagacatga ggtctatgga
     1021 cttcaagagc aacagtgctg tggcctggag caacaaatct gactttgcat gtgcaaacgc
     1081 cttcaacaac agcattattc cagaagacac cttcttcccc agcccagaaa gttcctgtga
     1141 tgtcaagctg gtcgagaaaa gctttgaaac agatacgaac ctaaactttc aaaacctgtc
     1201 agtgattggg ttccgaatcc tcctcctgaa agtggccggg tttaatctgc tcatgacgct
     1261 gcggctgtgg tccagctaaa gcggccgcgt cgacaatcaa atcggatccc gggcccgtcg
     1321 actgcagagg cctgcatgca agcttggtgt aatcatggtc atagctgttt cctgtgtgaa
     1381 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     1441 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     1501 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1561 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1621 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1681 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1741 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1801 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1861 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     1921 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     1981 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2041 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2101 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2161 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2221 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2281 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2341 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2401 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2461 attaaaaatg aagttttaaa tcaagcccaa tctgaataat gttacaacca attaaccaat
     2521 tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     2581 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     2641 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     2701 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     2761 acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca
     2821 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     2881 gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     2941 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     3001 ggatattctt ctaatacctg gaatgctgtt tttccgggga tcgcagtggt gagtaaccat
     3061 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     3121 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     3181 agaaacaact ctggcgcatc gggcttccca tacaagcgat agattgtcgc acctgattgc
     3241 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     3301 cgcggcctcg acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg
     3361 taagcagaca gttttattgt tcatgatgat atatttttat cttgtgcaat gtaacatcag
     3421 agattttgag acacgggcca gagctgca
//
