LOCUS       sequence-245030-        3421 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 3421)
  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 3421)
  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: TRBV12-5
FEATURES             Location/Qualifiers
     source          1..3421
                     /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"
     primer_bind     complement(1331..1347)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1331..1347)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1344..1366)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1355..1371
                     /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(1379..1385,1386..1403,1404..1409))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1424..1445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1562..1579)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1733..2321)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1813..1832)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2499..3308)
                     /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     3222..3241
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3300..3319
                     /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 ccatggccac caggctcctc tgctgtgtgg
      481 ttctttgtct cctgggagaa gagcttatag atgctagagt cacccagaca ccaaggcaca
      541 aggtgacaga gatgggacaa gaagtaacaa tgagatgtca gccaatttta ggccacaata
      601 ctgttttctg gtacagacag accatgatgc aaggactgga gttgctggct tacttccgca
      661 accgggctcc tctagatgat tcggggatgc cgaaggatcg attctcagca gagatgcctg
      721 atgcaacttt agccactctg aagatccagc cctcagaacc cagggactca gctgtgtatg
      781 agacctaagg tggtggtggt tctggttctg gttctggtgg tggttctggt ggtggtctct
      841 tgaccctgcc gtgtaccagc tgagagactc taaatccagt gacaagtctg tctgcctatt
      901 caccgatttt gattctcaaa caaatgtgtc acaaagtaag gattctgatg tgtatatcac
      961 agacaaaact gtgctagaca tgaggtctat ggacttcaag agcaacagtg ctgtggcctg
     1021 gagcaacaaa tctgactttg catgtgcaaa cgccttcaac aacagcatta ttccagaaga
     1081 caccttcttc cccagcccag aaagttcctg tgatgtcaag ctggtcgaga aaagctttga
     1141 aacagatacg aacctaaact ttcaaaacct gtcagtgatt gggttccgaa tcctcctcct
     1201 gaaagtggcc gggtttaatc tgctcatgac gctgcggctg tggtccagct aaagcggccg
     1261 cgtcgacaat caaatcggat cccgggcccg tcgactgcag aggcctgcat gcaagcttgg
     1321 tgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     1381 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     1441 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     1501 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     1561 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     1621 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1681 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1741 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1801 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1861 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1921 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1981 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2041 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2101 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2161 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2221 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2281 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2341 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2401 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaagcc
     2461 caatctgaat aatgttacaa ccaattaacc aattctgatt agaaaaactc atcgagcatc
     2521 aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt
     2581 ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat
     2641 cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa
     2701 ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa
     2761 agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa
     2821 tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg
     2881 cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact
     2941 gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct
     3001 gtttttccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc
     3061 ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta
     3121 acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc
     3181 ccatacaagc gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac
     3241 ccatataaat cagcatccat gttggaattt aatcgcggcc tcgacgtttc ccgttgaata
     3301 tggctcataa caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat
     3361 gatatatttt tatcttgtgc aatgtaacat cagagatttt gagacacggg ccagagctgc
     3421 a
//
