LOCUS Exported 3427 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION construction of TCR. ACCESSION . VERSION . KEYWORDS TRBV6-3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3427) 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 3427) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3427 /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" sig_peptide 453..518 /gene="pelB (fragment)" /product="leader peptide for secretion" /label=pelB signal sequence primer_bind complement(1337..1353) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1337..1353) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1350..1372) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1361..1377 /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(1385..1415) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1430..1451 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1568..1585) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1739..2327) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1819..1838) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2505..3314) /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 3228..3247 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 3306..3325 /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 ccatgaaata cctattgcct acggcagccg 481 ctggattgtt attactcgcg gcccagccgg ccatggccaa tgctggtgtc actcagaccc 541 caaaattccg ggtcctgaag acaggacaga gcatgacact gctgtgtgcc caggatatga 601 accatgaata catgtactgg tatcgacaag acccaggcat ggggctgagg ctgattcatt 661 actcagttgg tgagggtaca actgccaaag gagaggtccc tgatggctac aatgtctcca 721 gattaaaaaa acagaatttc ctgctggggt tggagtcggc tgctccctcc caaacatctg 781 tgtacgagac ctaaggtggt ggtggttctg gttctggttc tggtggtggt tctggtggtg 841 gtctcttgac cctgccgtgt accagctgag agactctaaa tccagtgaca agtctgtctg 901 cctattcacc gattttgatt ctcaaacaaa tgtgtcacaa agtaaggatt ctgatgtgta 961 tatcacagac aaaactgtgc tagacatgag gtctatggac ttcaagagca acagtgctgt 1021 ggcctggagc aacaaatctg actttgcatg tgcaaacgcc ttcaacaaca gcattattcc 1081 agaagacacc ttcttcccca gcccagaaag ttcctgtgat gtcaagctgg tcgagaaaag 1141 ctttgaaaca gatacgaacc taaactttca aaacctgtca gtgattgggt tccgaatcct 1201 cctcctgaaa gtggccgggt ttaatctgct catgacgctg cggctgtggt ccagctaaag 1261 cggccgcgtc gacaatcaaa tcggatcccg ggcccgtcga ctgcagaggc ctgcatgcaa 1321 gcttggtgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 1381 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 1441 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 1501 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 1561 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 1621 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 1681 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 1741 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 1801 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 1861 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 1921 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 1981 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 2041 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 2101 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 2161 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct 2221 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 2281 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 2341 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 2401 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 2461 caagcccaat ctgaataatg ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg 2521 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa 2581 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc 2641 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg 2701 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat 2761 ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca 2821 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga 2881 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg 2941 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg 3001 aatgctgttt ttccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata 3061 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca 3121 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg 3181 ggcttcccat acaagcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat 3241 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctcga cgtttcccgt 3301 tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt 3361 catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga cacgggccag 3421 agctgca //