LOCUS Exported 6861 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDONR223-KDR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6861) 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 6861) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6861 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 23..109 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 158..180 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 172..189 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 173..189 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 205..304 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(4376..4475) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(4492..4511) /label=T7 /note="T7 promoter, forward primer" promoter complement(4493..4511) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4516..4532) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4516..4532) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 4963..5754 /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 5847..6435 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6336..6355 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6589..6606 /label=L4440 /note="L4440 vector, forward primer" terminator 6765..6792 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" ORIGIN 1 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac 61 agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag 121 ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg ttgtaaaacg 181 acggccagtc ttaagctcgg gccccaaata atgattttat tttgactgat agtgacctgt 241 tcgttgcaac aaattgatga gcaatgcttt tttataatgc caactttgta caaaaaagtt 301 ggcaccatgc agagcaaggt gctgctggcc gtcgccctgt ggctctgcgt ggagacccgg 361 gccgcctctg tgggtttgcc tagtgtttct cttgatctgc ccaggctcag catacaaaaa 421 gacatactta caattaaggc taatacaact cttcaaatta cttgcagggg acagagggac 481 ttggactggc tttggcccaa taatcagagt ggcagtgagc aaagggtgga ggtgactgag 541 tgcagcgatg gcctcttctg taagacactc acaattccaa aagtgatcgg aaatgacact 601 ggagcctaca agtgcttcta ccgggaaact gacttggcct cggtcattta tgtctatgtt 661 caagattaca gatctccatt tattgcttct gttagtgacc aacatggagt cgtgtacatt 721 actgagaaca aaaacaaaac tgtggtgatt ccatgtctcg ggtccatttc aaatctcaac 781 gtgtcacttt gtgcaagata cccagaaaag agatttgttc ctgatggtaa cagaatttcc 841 tgggacagca agaagggctt tactattccc agctacatga tcagctatgc tggcatggtc 901 ttctgtgaag caaaaattaa tgatgaaagt taccagtcta ttatgtacat agttgtcgtt 961 gtagggtata ggatttatga tgtggttctg agtccgtctc atggaattga actatctgtt 1021 ggagaaaagc ttgtcttaaa ttgtacagca agaactgaac taaatgtggg gattgacttc 1081 aactgggaat acccttcttc gaagcatcag cataagaaac ttgtaaaccg agacctaaaa 1141 acccagtctg ggagtgagat gaagaaattt ttgagcacct taactataga tggtataacc 1201 cggagtgacc aaggattgta cacctgtgca gcatccagtg ggctgatgac caagaagaac 1261 agcacatttg tcagggtcca tgaaaaacct tttgttgctt ttggaagtgg catggaatct 1321 ctggtggaag ccacggtggg ggagcgtgtc agaatccctg cgaagtacct tggttaccca 1381 cccccagaaa taaaatggta taaaaatgga ataccccttg agtccaatca cacaattaaa 1441 gcggggcatg tactgacgat tatggaagtg agtgaaagag acacaggaaa ttacactgtc 1501 atccttacca atcccatttc aaaggagaag cagagccatg tggtctctct ggttgtgtat 1561 gtcccacccc agattggtga gaaatctcta atctctcctg tggattccta ccagtacggc 1621 accactcaaa cgctgacatg tacggtctat gccattcctc ccccgcatca catccactgg 1681 tattggcagt tggaggaaga gtgcgccaac gagcccagcc aagctgtctc agtgacaaac 1741 ccataccctt gtgaagaatg gagaagtgtg gaggacttcc agggaggaaa taaaattgaa 1801 gttaataaaa atcaatttgc tctaattgaa ggaaaaaaca aaactgtaag tacccttgtt 1861 atccaagcgg caaatgtgtc agctttgtac aaatgtgaag cggtcaacaa agtcgggaga 1921 ggagagaggg tgatctcctt ccacgtgacc aggggtcctg aaattacttt gcaacctgac 1981 atgcagccca ctgagcagga gagcgtgtct ttgtggtgca ctgcagacag atctacgttt 2041 gagaacctca catggtacaa gcttggccca cagcctctgc caatccatgt gggagagttg 2101 cccacacctg tttgcaagaa cttggatact ctttggaaat tgaatgccac catgttctct 2161 aatagcacaa atgacatttt gatcatggag cttaagaatg catccttgca ggaccaagga 2221 gactatgtct gccttgctca agacaggaag accaagaaaa gacattgcgt ggtcaggcag 2281 ctcacagtcc tagagcgtgt ggcacccacg atcacaggaa acctggagaa tcagacgaca 2341 agtattgggg aaagcatcga agtctcatgc acggcatctg ggaatccccc tccacagatc 2401 atgtggttta aagataatga gacccttgta gaagactcag gcattgtatt gaaggatggg 2461 aaccggaacc tcactatccg cagagtgagg aaggaggacg aaggcctcta cacctgccag 2521 gcatgcagtg ttcttggctg tgcaaaagtg gaggcatttt tcataataga aggtgcccag 2581 gaaaagacga acttggaaat cattattcta gtaggcacgg cggtgattgc catgttcttc 2641 tggctacttc ttgtcatcat cctacggacc gttaagcggg ccaatggagg ggaactgaag 2701 acaggctact tgtccatcgt catggatcca gatgaactcc cattggatga acattgtgaa 2761 cgactgcctt atgatgccag caaatgggaa ttccccagag accggctgaa gctaggtaag 2821 cctcttggcc gtggtgcctt tggccaagtg attgaagcag atgcctttgg aattgacaag 2881 acagcaactt gcaggacagt agcagtcaaa atgttgaaag aaggagcaac acacagtgag 2941 catcgagctc tcatgtctga actcaagatc ctcattcata ttggtcacca tctcaatgtg 3001 gtcaaccttc taggtgcctg taccaagcca ggagggccac tcatggtgat tgtggaattc 3061 tgcaaatttg gaaacctgtc cacttacctg aggagcaaga gaaatgaatt tgtcccctac 3121 aagaccaaag gggcacgatt ccgtcaaggg aaagactacg ttggagcaat ccctgtggat 3181 ctgaaacggc gcttggacag catcaccagt agccagagct cagccagctc tggatttgtg 3241 gaggagaagt ccctcagtga tgtagaagaa gaggaagctc ctgaagatct gtataaggac 3301 ttcctgacct tggagcatct catctgttac agcttccaag tggctaaggg catggagttc 3361 ttggcatcgc gaaagtgtat ccacagggac ctggcggcac gaaatatcct cttatcggag 3421 aagaacgtgg ttaaaatctg tgactttggc ttggcccggg atatttataa agatccagat 3481 tatgtcagaa aaggagatgc tcgcctccct ttgaaatgga tggccccaga aacaattttt 3541 gacagagtgt acacaatcca gagtgacgtc tggtcttttg gtgttttgct gtgggaaata 3601 ttttccttag gtgcttctcc atatcctggg gtaaagattg atgaagaatt ttgtaggcga 3661 ttgaaagaag gaactagaat gagggcccct gattatacta caccagaaat gtaccagacc 3721 atgctggact gctggcacgg ggagcccagt cagagaccca cgttttcaga gttggtggaa 3781 catttgggaa atctcttgca agctaatgct cagcaggatg gcaaagacta cattgttctt 3841 ccgatatcag agactttgag catggaagag gattctggac tctctctgcc tacctcacct 3901 gtttcctgta tggaggagga ggaagtatgt gaccccaaat tccattatga caacacagca 3961 ggaatcagtc agtatctgca gaacagtaag cgaaagagcc ggcctgtgag tgtaaaaaca 4021 tttgaagata tcccgttaga agaaccagaa gtaaaagtaa tcccagatga caaccagacg 4081 gacagtggta tggttcttgc ctcagaagag ctgaaaactt tggaagacag aaccaaatta 4141 tctccatctt ttggtggaat ggtgcccagc aaaagcaggg agtctgtggc atctgaaggc 4201 tcaaaccaga caagcggcta ccagtccgga tatcactccg atgacacaga caccaccgtg 4261 tactccagtg aggaagcaga acttttaaag ctgatagaga ttggagtgca aaccggtagc 4321 acagcccaga ttctccagcc tgactcgggg accacactga gctctcctcc tgttttgcca 4381 actttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa 4441 caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt 4501 gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc 4561 tgatgttaca ttgcacaaga taaaaatata tcatcatgcc tcctctagac cagccaggac 4621 agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg 4681 atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag 4741 cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca 4801 gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca 4861 tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat 4921 gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc 4981 gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg 5041 acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt 5101 gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg 5161 atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta 5221 gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa 5281 ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg 5341 atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta 5401 ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta 5461 aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta 5521 gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat 5581 gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa 5641 gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg 5701 gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg 5761 agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac 5821 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 5881 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 5941 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta 6001 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 6061 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 6121 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 6181 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta 6241 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 6301 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 6361 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 6421 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 6481 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc 6541 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg 6601 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt 6661 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca 6721 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca 6781 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc 6841 ctccgggccg ttgcttcaca a //