LOCUS Exported 3531 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION construction of TCR. ACCESSION . VERSION . KEYWORDS TRAV13-1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3531) 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 3531) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3531 /organism="synthetic DNA construct" /mol_type="other DNA" 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(1441..1457) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1441..1457) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1454..1476) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1465..1481 /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(1489..1519) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1534..1555 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1672..1689) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1843..2431) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1923..1942) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2609..3418) /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 3332..3351 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 3410..3429 /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 ggcccatgac atccattcga gctgtattta 1081 tattcctgtg gctgcagctg gacttggtga atggagagaa tgtggagcag catccttcaa 1141 ccctgagtgt ccaggaggga gacagcgctg ttatcaagtg tacttattca gacagtgcct 1201 caaactactt cccttggtat aagcaagaac ttggaaaagg acctcagctt attatagaca 1261 ttcgttcaaa tgtgggcgaa aagaaagacc aacgaattgc tgttacattg aacaagacag 1321 ccaaacattt ctccctgcac atcacagaga cacaacctga agactcggct gtctactgag 1381 accatcggat cccgggcccg tcgactgcag aggcctgcat gcaagcttgg tgtaatcatg 1441 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 1501 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 1561 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 1621 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 1681 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 1741 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 1801 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 1861 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 1921 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 1981 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 2041 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 2101 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 2161 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 2221 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 2281 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 2341 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 2401 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 2461 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 2521 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaagcc caatctgaat 2581 aatgttacaa ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact 2641 gcaatttatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg 2701 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga 2761 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat 2821 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca 2881 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat 2941 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt 3001 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat 3061 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gtttttccgg 3121 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg 3181 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg 3241 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaagc 3301 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat 3361 cagcatccat gttggaattt aatcgcggcc tcgacgtttc ccgttgaata tggctcataa 3421 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt 3481 tatcttgtgc aatgtaacat cagagatttt gagacacggg ccagagctgc a //