LOCUS Exported 3534 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION construction of TCR. ACCESSION . VERSION . KEYWORDS TRAV3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3534) 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 3534) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3534 /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(1444..1460) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1444..1460) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1457..1479) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1468..1484 /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(1492..1522) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1537..1558 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1675..1692) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1846..2434) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1926..1945) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2612..3421) /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 3335..3354 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 3413..3432 /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 ggcccatggc ctctgcaccc atctcgatgc 1081 ttgcgatgct cttcacattg agtgggctga gagctcagtc agtggctcag ccggaagatc 1141 aggtcaacgt tgctgaaggg aatcctctga ctgtgaaatg cacctattca gtctctggaa 1201 acccttatct tttttggtat gttcaatacc ccaaccgagg cctccagttc cttctgaaat 1261 acatcacagg ggataacctg gttaaaggca gctatggctt tgaagctgaa tttaacaaga 1321 gccaaacctc cttccacctg aagaaaccat ctgcccttgt gagcgactcc gctttgtact 1381 gagaccatcg gatcccgggc ccgtcgactg cagaggcctg catgcaagct tggtgtaatc 1441 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 1501 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 1561 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 1621 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 1681 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 1741 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 1801 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 1861 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 1921 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 1981 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 2041 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 2101 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 2161 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 2221 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 2281 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 2341 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 2401 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 2461 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 2521 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa gcccaatctg 2581 aataatgtta caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa 2641 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta 2701 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg 2761 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt 2821 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat 2881 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg 2941 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc 3001 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg 3061 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgtttttc 3121 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg 3181 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat 3241 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca 3301 agcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata 3361 aatcagcatc catgttggaa tttaatcgcg gcctcgacgt ttcccgttga atatggctca 3421 taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat 3481 ttttatcttg tgcaatgtaa catcagagat tttgagacac gggccagagc tgca //