LOCUS Exported 3418 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION construction of TCR. ACCESSION . VERSION . KEYWORDS TRBV6-4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3418) 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 3418) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3418 /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" primer_bind complement(1328..1344) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1328..1344) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1341..1363) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1352..1368 /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(1376..1406) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1421..1442 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1559..1576) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1730..2318) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1810..1829) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2496..3305) /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 3219..3238 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 3297..3316 /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 ccatgagaat caggctcctg tgctgtgtgg 481 ccttttctct cctgtgggca ggtccagtga ttgctgggat cacccaggca ccaacatctc 541 agatcctggc agcaggacgg cgcatgacac tgagatgtac ccaggatatg agacataatg 601 ccatgtactg gtatagacaa gatctaggac tggggctaag gctcatccat tattcaaata 661 ctgcaggtac cactggcaaa ggagaagtcc ctgatggtta tagtgtctcc agagcaaaca 721 cagatgattt ccccctcacg ttggcgtctg ctgtaccctc tcagacatct gtgtacgaga 781 cctaaggtgg tggtggttct ggttctggtt ctggtggtgg ttctggtggt ggtctcttga 841 ccctgccgtg taccagctga gagactctaa atccagtgac aagtctgtct gcctattcac 901 cgattttgat tctcaaacaa atgtgtcaca aagtaaggat tctgatgtgt atatcacaga 961 caaaactgtg ctagacatga ggtctatgga cttcaagagc aacagtgctg tggcctggag 1021 caacaaatct gactttgcat gtgcaaacgc cttcaacaac agcattattc cagaagacac 1081 cttcttcccc agcccagaaa gttcctgtga tgtcaagctg gtcgagaaaa gctttgaaac 1141 agatacgaac ctaaactttc aaaacctgtc agtgattggg ttccgaatcc tcctcctgaa 1201 agtggccggg tttaatctgc tcatgacgct gcggctgtgg tccagctaaa gcggccgcgt 1261 cgacaatcaa atcggatccc gggcccgtcg actgcagagg cctgcatgca agcttggtgt 1321 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 1381 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 1441 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 1501 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 1561 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 1621 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 1681 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 1741 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 1801 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 1861 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 1921 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 1981 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 2041 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 2101 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 2161 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 2221 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 2281 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 2341 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 2401 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaagcccaa 2461 tctgaataat gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa 2521 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc 2581 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg 2641 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata 2701 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt 2761 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca 2821 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga 2881 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc 2941 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt 3001 tttccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg 3061 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca 3121 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca 3181 tacaagcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca 3241 tataaatcag catccatgtt ggaatttaat cgcggcctcg acgtttcccg ttgaatatgg 3301 ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat 3361 atatttttat cttgtgcaat gtaacatcag agattttgag acacgggcca gagctgca //