LOCUS       sequence-244187-        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: TRBV2
FEATURES             Location/Qualifiers
     source          1..3421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     179..201
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     regulatory      262..271
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1146..1162)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1146..1162)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1159..1181)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1170..1186
                     /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(1194..1200,1201..1218,1219..1224))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1239..1260
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1377..1394)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1548..2136)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1628..1647)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2314..3123)
                     /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     3037..3056
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3115..3134
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(3265..3287)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3387..3406
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg
       61 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
      121 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
      181 cagtcacgac gttgtaaaac gacggccaga gaattcgagc tcggtacctc gcgaatacat
      241 ctagatgggg atccaccggt cgccaccatg gatacctggc tcgtatgctg ggcaattttt
      301 agtctcttga aagcaggact cacagaacct gaagtcaccc agactcccag ccatcaggtc
      361 acacagatgg gacaggaagt gatcttgcgc tgtgtcccca tctctaatca cttatacttc
      421 tattggtaca gacaaatctt ggggcagaaa gtcgagtttc tggtttcctt ttataataat
      481 gaaatctcag agaagtctga aatattcgat gatcaattct cagttgaaag gcctgatgga
      541 tcaaatttca ctctgaagat ccggtccaca aagctggagg actcagccat gtacgagacc
      601 taaggtggtg gtggttctgg ttctggttct ggtggtggtt ctggtggtgg tctcttgacc
      661 ctgccgtgta ccagctgaga gactctaaat ccagtgacaa gtctgtctgc ctattcaccg
      721 attttgattc tcaaacaaat gtgtcacaaa gtaaggattc tgatgtgtat atcacagaca
      781 aaactgtgct agacatgagg tctatggact tcaagagcaa cagtgctgtg gcctggagca
      841 acaaatctga ctttgcatgt gcaaacgcct tcaacaacag cattattcca gaagacacct
      901 tcttccccag cccagaaagt tcctgtgatg tcaagctggt cgagaaaagc tttgaaacag
      961 atacgaacct aaactttcaa aacctgtcag tgattgggtt ccgaatcctc ctcctgaaag
     1021 tggccgggtt taatctgctc atgacgctgc ggctgtggtc cagctaaagc ggccgcgtcg
     1081 acaatcaaat cggatcccgg gcccgtcgac tgcagaggcc tgcatgcaag cttggtgtaa
     1141 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     1201 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     1261 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     1321 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     1381 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     1441 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     1501 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1561 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1621 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1681 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1741 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1801 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1861 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     1921 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     1981 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2041 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2101 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2161 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2221 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aagcccaatc
     2281 tgaataatgt tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg
     2341 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     2401 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     2461 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     2521 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt
     2581 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     2641 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     2701 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     2761 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     2821 tccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     2881 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     2941 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     3001 caagcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     3061 taaatcagca tccatgttgg aatttaatcg cggcctcgac gtttcccgtt gaatatggct
     3121 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat
     3181 atttttatct tgtgcaatgt aacatcagag attttgagac acgggccaga gctgcatcgc
     3241 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     3301 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     3361 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     3421 t
//
