LOCUS Exported 4044 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDONR223-PRKAR2B SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4044) 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 4044) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4044 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 72..88 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 72..88 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 93..112 /label=T7 /note="T7 promoter, forward primer" promoter 93..111 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 129..228 /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(1483..1582) /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1598..1615) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(1598..1614) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1607..1629) /label=M13/pUC Forward /note="In lacZ gene" terminator 1678..1764 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 1856..1883 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" primer_bind complement(2042..2059) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2213..2801) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2293..2312) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2894..3685) /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" ORIGIN 1 ggaggcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacgg 61 gccagagctg ccaggaaaca gctatgacca tgtaatacga ctcactatag gggatatcag 121 ctggatggca aataatgatt ttattttgac tgatagtgac ctgttcgttg caacaaattg 181 ataagcaatg ctttcttata atgccaactt tgtacaagaa agttgggcat gcagtgggtt 241 caacaatatc catgttcgtt ccaaacaggg caactaactg ttcttcatag gtagcgatgt 301 tccttttcat aatttccatg caaggtccca gaagcctttc aaatgcttgc acatccattg 361 ctaaacattt gacagtccca atggcgtggg cagaagctgc tcgaggtttg ttagttacca 421 gggcaagctc tccaaagtac tgtccccgcg agcatcgagc gatttctact gcaccattct 481 cttccacttc tgatttaccc tttcttttca tagtaatttt cacttctcca gattctacaa 541 tgaaaaaaga atcagccgaa tctccctgag caatgatttg ttctccatcg ttgtatactt 601 tggtgcctat cacatctact actttcaggc gttcagaaaa ctccaaagat ttaaggaatg 661 gcagtgactc aataaagctt tcatacattt ttctcttttt ggcattgttt ttcacaatta 721 ttctcctgaa ggttaccctg tccaaacccc acagagcacc aggagaggta gcagtgattg 781 tagctgctct gggtgtattg tacattaagg ccagttcgcc gaaactccca cgattatcat 841 agttaccaac acatcttcca acaccatcac atttcacata aatatcaaat gtgcctctat 901 caattacata aaagttgtca ccatcgtcac cttgatcaat tacatgctcc ccatctttga 961 ccaatttttc aaacatggca tctaatactt gagacatctg ctccggatcc agattcttaa 1021 acagcaggat gtctttgcaa gcctcttgca acctatttct ttgatcatca gtttttggat 1081 gtataatcct ggactctgca tcatcttctt cttcatcagg attataagct tctgcacata 1141 ctgaggcacg ccttgtgaat cggtttatta ctggagcatt gaacgcccct gcgtccgcgg 1201 gcgccgcctc ctcctcctcc ccgtcctcgg agtcggactg catgggctcc tcggcgaagt 1261 tgaccccctt gctgggggtg ccgcccccgg cagcggcgcc caggtccccc caggtcctgc 1321 cctcatggca gaagcgcgcg gtgcctttgc gctcgttctc ctgctgcagg cgggtgaagt 1381 gctgcagagc gaactccagc aggtccgcgg gctggtgcct cagcacctcc accgtgaagc 1441 cctgcagcag ctccgtcagt cccgccggga tctcgatgct catgccaact tttttgtaca 1501 aagttggcat tataaaaaag cattgctcat caatttgttg caacgaacag gtcactatca 1561 gtcaaaataa aatcattatt tggggcccga gcttaagact ggccgtcgtt ttacaacgtc 1621 gtgactggga aaacatccat gctagcgtta acgcgagagt agggaactgc caggcatcaa 1681 ataaaacgaa aggctcagtc ggaagactgg gcctttcgtt ttatctgttg tttgtcggtg 1741 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgt gaagcaacgg 1801 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaac taagcagaag 1861 gccatcctga cggatggcct ttttgcgttt ctacaaactc ttcctggcta gcggtacgcg 1921 tattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 1981 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 2041 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 2101 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 2161 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 2221 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 2281 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 2341 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 2401 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 2461 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 2521 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 2581 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 2641 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 2701 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 2761 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 2821 ctacggggtc tgacgctcag tggaacgacg cgtaactcac gttaagggat tttggtcatg 2881 ggtggctcga gggttatttg ccgactacct tggtgatctc gcctttcacg tagtggacaa 2941 attcttccaa ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca agataagcct 3001 gtctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt gcccagtcgg 3061 cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg cgggacaacg 3121 taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg 3181 tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg 3241 ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa 3301 tgtcgatcgt ggctggctcg aagatacctg caagaatgtc attgcgctgc cattctccaa 3361 attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg 3421 tgacttctac agcgcggaga atctcgctct ctccagggga agccgaagtt tccaaaaggt 3481 cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat 3541 caatatcact gtgtggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca 3601 gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata 3661 cttcggcgat caccgcttcc ctcataatgt ttaactttgt tttagggcga ctgccctgct 3721 gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa cgcgcttgct 3781 gcttggatgc ccgaggcata gactgtaccc caaaaaaaca gtcataacaa gccatgaaaa 3841 ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc 3901 gcatacgcta cttgcattac agcttacgaa ccgaacaggc ttatgtccac tgggttcgtg 3961 ccttcatccg tttccacggt gtgcgtcacc cggcaacctt gggtagcagc gaagtcgagg 4021 catttctgtc ctggctggtc taga //