LOCUS Exported 6960 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDONR223-MET SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6960) AUTHORS Johannessen CM, Boehm JS, Kim SY, Thomas SR, Wardwell L, Johnson LA, Emery CM, Stransky N, Cogdill AP, Barretina J, Caponigro G, Hieronymus H, Murray RR, Salehi-Ashtiani K, Hill DE, Vidal M, Zhao JJ, Yang X, Alkan O, Kim S, Harris JL, Wilson CJ, Myer VE, Finan PM, Root DE, Roberts TM, Golub T, Flaherty KT, Dummer R, Weber BL, Sellers WR, Schlegel R, Wargo JA, Hahn WC, Garraway LA TITLE COT drives resistance to RAF inhibition through MAP kinase pathway reactivation. JOURNAL Nature. 2010 Nov 24. ():. PUBMED 21107320 REFERENCE 2 (bases 1 to 6960) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6960 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 54..140 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 189..211 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 203..220 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 204..220 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 236..335 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(4506..4605) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(4622..4641) /label=T7 /note="T7 promoter, forward primer" promoter complement(4623..4641) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4646..4662) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4646..4662) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 5093..5884 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin 5977..6565 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6466..6485 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6719..6736 /label=L4440 /note="L4440 vector, forward primer" terminator 6895..6922 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" ORIGIN 1 gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc 61 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac 121 tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg 181 gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccccaaat 241 aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgatg agcaatgctt 301 ttttataatg ccaactttgt acaaaaaagt tggcaccatg aaggcccccg ctgtgcttgc 361 acctggcatc ctcgtgctcc tgtttacctt ggtgcagagg agcaatgggg agtgtaaaga 421 ggcactagca aagtccgaga tgaatgtgaa tatgaagtat cagcttccca acttcaccgc 481 ggaaacaccc atccagaatg tcattctaca tgagcatcac attttccttg gtgccactaa 541 ctacatttat gttttaaatg aggaagacct tcagaaggtt gctgagtaca agactgggcc 601 tgtgctggaa cacccagatt gtttcccatg tcaggactgc agcagcaaag ccaatttatc 661 aggaggtgtt tggaaagata acatcaacat ggctctagtt gtcgacacct actatgatga 721 tcaactcatt agctgtggca gcgtcaacag agggacctgc cagcgacatg tctttcccca 781 caatcatact gctgacatac agtcggaggt tcactgcata ttctccccac agatagaaga 841 gcccagccag tgtcctgact gtgtggtgag cgccctggga gccaaagtcc tttcatctgt 901 aaaggaccgg ttcatcaact tctttgtagg caataccata aattcttctt atttcccaga 961 tcatccattg cattcgatat cagtgagaag gctaaaggaa acgaaagatg gttttatgtt 1021 tttgacggac cagtcctaca ttgatgtttt acctgagttc agagattctt accccattaa 1081 gtatgtccat gcctttgaaa gcaacaattt tatttacttc ttgacggtcc aaagggaaac 1141 tctagatgct cagacttttc acacaagaat aatcaggttc tgttccataa actctggatt 1201 gcattcctac atggaaatgc ctctggagtg tattctcaca gaaaagagaa aaaagagatc 1261 cacaaagaag gaagtgttta atatacttca ggctgcgtat gtcagcaagc ctggggccca 1321 gcttgctaga caaataggag ccagcctgaa tgatgacatt cttttcgggg tgttcgcaca 1381 aagcaagcca gattctgccg aaccaatgga tcgatctgcc atgtgtgcat tccctatcaa 1441 atatgtcaac gacttcttca acaagatcgt caacaaaaac aatgtgagat gtctccagca 1501 tttttacgga cccaatcatg agcactgctt taataggaca cttctgagaa attcatcagg 1561 ctgtgaagcg cgccgtgatg aatatcgaac agagtttacc acagctttgc agcgcgttga 1621 cttattcatg ggtcaattca gcgaagtcct cttaacatct atatccacct tcattaaagg 1681 agacctcacc atagctaatc ttgggacatc agagggtcgc ttcatgcagg ttgtggtttc 1741 tcgatcagga ccatcaaccc ctcatgtgaa ttttctcctg gactcccatc cagtgtctcc 1801 agaagtgatt gtggagcata cattaaacca aaatggctac acactggtta tcactgggaa 1861 gaagatcacg aagatcccat tgaatggctt gggctgcaga catttccagt cctgcagtca 1921 atgcctctct gccccaccct ttgttcagtg tggctggtgc cacgacaaat gtgtgcgatc 1981 ggaggaatgc ctgagcggga catggactca acagatctgt ctgcctgcaa tctacaaggt 2041 tttcccaaat agtgcacccc ttgaaggagg gacaaggctg accatatgtg gctgggactt 2101 tggatttcgg aggaataata aatttgattt aaagaaaact agagttctcc ttggaaatga 2161 gagctgcacc ttgactttaa gtgagagcac gatgaataca ttgaaatgca cagttggtcc 2221 tgccatgaat aagcatttca atatgtccat aattatttca aatggccacg ggacaacaca 2281 gtacagtaca ttctcctatg tggatcctgt aataacaagt atttcgccga aatacggtcc 2341 tatggctggt ggcactttac ttactttaac tggaaattac ctaaacagtg ggaattctag 2401 acacatttca attggtggaa aaacatgtac tttaaaaagt gtgtcaaaca atattcttga 2461 atgttatacc ccagcccaaa ccatttcaac tgagtttgct gttaaattga aaattgactt 2521 agccaaccga gagacaagca tcttcagtta ccgtgaagat cccattgtct atgaaattca 2581 tccaaccaaa tcttttatta gtggtgggag cacaataaca ggtgttggga aaaacctgaa 2641 ttcagttagt gtcccgagaa tggtcataaa tgtgcatgaa gcaggaagga actttacagt 2701 ggcatgtcaa catcgctcta attcagagat aatctgttgt accactcctt ccctgcaaca 2761 gctgaatctg caactccccc tgaaaaccaa agcctttttc atgttagatg ggatcctttc 2821 caaatacttt gatctcattt atgtacataa tcctgtgttt aagccttttg aaaagccagt 2881 gatgatctca atgggcaatg aaaatgtact ggaaattaag ggaaatgata ttgaccctga 2941 agcagttaaa ggtgaagtgt taaaagttgg aaataagagc tgtgagaata tacacttaca 3001 ttctgaagcc gttttatgca cggtccccaa tgacctgctg aaattgaaca gcgagctaaa 3061 tatagagtgg aagcaagcaa tttcttcaac cgtccttgga aaagtaatag ttcaaccaga 3121 tcagaatttc acaggattga ttgctggtgt tgtctcaata tcaacagcac tgttattact 3181 acttgggttt ttcctgtggc tgaaaaagag aaagcaaatt aaagatctgg gcagtgaatt 3241 agttcgctac gatgcaagag tacacactcc tcatttggat aggcttgtaa gtgcccgaag 3301 tgtaagccca actacagaaa tggtttcaaa tgaatctgta gactaccgag ctacttttcc 3361 agaagatcag tttcctaatt catctcagaa cggttcatgc cgacaagtgc agtatcctct 3421 gacagacatg tcccccatcc taactagtgg ggactctgat atatccagtc cattactgca 3481 aaatactgtc cacattgacc tcagtgctct aaatccagag ctggtccagg cagtgcagca 3541 tgtagtgatt gggcccagta gcctgattgt gcatttcaat gaagtcatag gaagagggca 3601 ttttggttgt gtatatcatg ggactttgtt ggacaatgat ggcaagaaaa ttcactgtgc 3661 tgtgaaatcc ttgaacagaa tcactgacat aggagaagtt tcccaatttc tgaccgaggg 3721 aatcatcatg aaagatttta gtcatcccaa tgtcctctcg ctcctgggaa tctgcctgcg 3781 aagtgaaggg tctccgctgg tggtcctacc atacatgaaa catggagatc ttcgaaattt 3841 cattcgaaat gagactcata atccaactgt aaaagatctt attggctttg gtcttcaagt 3901 agccaaaggc atgaaatatc ttgcaagcaa aaagtttgtc cacagagact tggctgcaag 3961 aaactgtatg ctggatgaaa aattcacagt caaggttgct gattttggtc ttgccagaga 4021 catgtatgat aaagaatact atagtgtaca caacaaaaca ggtgcaaagc tgccagtgaa 4081 gtggatggct ttggaaagtc tgcaaactca aaagtttacc accaagtcag atgtgtggtc 4141 ctttggcgtg ctcctctggg agctgatgac aagaggagcc ccaccttatc ctgacgtaaa 4201 cacctttgat ataactgttt acttgttgca agggagaaga ctcctacaac ccgaatactg 4261 cccagacccc ttatatgaag taatgctaaa atgctggcac cctaaagccg aaatgcgccc 4321 atccttttct gaactggtgt cccggatatc agcgatcttc tctactttca ttggggagca 4381 ctatgtccat gtgaacgcta cttatgtgaa cgtaaaatgt gtcgctccgt atccttctct 4441 gttgtcatca gaagataacg ctgatgatga ggtggacaca cgaccagcct ccttctggga 4501 gacatcacca actttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg 4561 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat 4621 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc 4681 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgcc tcctctagac 4741 cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc 4801 gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta 4861 atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg 4921 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat 4981 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag 5041 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga 5101 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca 5161 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa 5221 gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg 5281 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct 5341 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc 5401 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga 5461 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt 5521 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt 5581 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga 5641 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc 5701 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt 5761 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc 5821 agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa 5881 ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg 5941 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt 6001 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca 6061 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac 6121 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac 6181 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct 6241 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg 6301 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca 6361 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt 6421 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta 6481 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc 6541 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc 6601 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa 6661 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag 6721 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg 6781 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga 6841 gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg 6901 ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct //