LOCUS Exported 3756 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDONR223-PBK SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3756) 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 3756) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3756 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 133..152 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 386..403 /label=L4440 /note="L4440 vector, forward primer" terminator 562..589 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 681..767 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 816..838 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 830..847 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 831..847 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 863..962 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(1929..2028) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(2045..2064) /label=T7 /note="T7 promoter, forward primer" promoter complement(2046..2064) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2069..2085) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2069..2085) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 2516..3307 /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 3400..232 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga 61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag 121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg 181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag 241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc 301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc 361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc 421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag 481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca 541 ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga 601 tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg 661 ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag 721 ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt 781 ccctactctc gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac 841 ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag tgacctgttc 901 gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca aaaaagttgg 961 catggaaggg atcagtaatt tcaagacacc aagcaaatta tcagaaaaaa agaaatctgt 1021 attatgttca actccaacta taaatatccc ggcctctcca tttatgcaga agcttggctt 1081 tggtactggg gtaaatgtgt acctaatgaa aagatctcca agaggtttgt ctcattctcc 1141 ttgggctgta aaaaagatta atcctatatg taatgatcat tatcgaagtg tgtatcaaaa 1201 gagactaatg gatgaagcta agattttgaa aagccttcat catccaaaca ttgttggtta 1261 tcgtgctttt actgaagcca atgatggcag tctgtgtctt gctatggaat atggaggtga 1321 aaagtctcta aatgacttaa tagaagaacg atataaagcc agccaagatc cttttccagc 1381 agccataatt ttaaaagttg ctttgaatat ggcaagaggg ttaaagtatc tgcaccaaga 1441 aaagaaactg cttcatggag acataaagtc ttcaaatgtt gtaattaaag gcgattttga 1501 aacaattaaa atctgtgatg taggagtctc tctaccactg gatgaaaata tgactgtgac 1561 tgaccctgag gcttgttaca ttggcacaga gccatggaaa cccaaagaag ctgtggagga 1621 gaatggtgtt attactgaca aggcagacat atttgccttt ggccttactt tgtgggaaat 1681 gatgacttta tcgattccac acattaatct ttcaaatgat gatgatgatg aagataaaac 1741 ttttgatgaa agtgattttg atgatgaagc atactatgca gcgttgggaa ctaggccacc 1801 tattaatatg gaagaactgg atgaatcata ccagaaagta attgaactct tctctgtatg 1861 cactaatgaa gaccctaaag atcgtccttc tgctgcacac attgttgaag ctctggaaac 1921 agatgtctac ccaactttct tgtacaaagt tggcattata agaaagcatt gcttatcaat 1981 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga 2041 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt 2101 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gcctcctcta 2161 gaccagccag gacagaaatg cctcgacttc gctgctaccc aaggttgccg ggtgacgcac 2221 accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct 2281 gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg 2341 tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc 2401 tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg 2461 gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa acattatgag 2521 ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg 2581 ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct 2641 gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac 2701 gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat 2761 tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc 2821 agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt 2881 cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag 2941 cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct 3001 atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga 3061 tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat 3121 cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc 3181 cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg 3241 cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg 3301 caaataaccc tcgagccacc catgaccaaa atcccttaac gtgagttacg cgtcgttcca 3361 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg 3421 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga 3481 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa 3541 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc 3601 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg 3661 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac 3721 ggggggttcg tgcacacagc ccagcttgga gcgaac //