LOCUS Exported 7650 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDONR223-ALK SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7650) 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 7650) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7650 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 149..176 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 268..354 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 403..425 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 417..434 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 418..434 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 450..549 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(5410..5509) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(5526..5545) /label=T7 /note="T7 promoter, forward primer" promoter complement(5527..5545) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5550..5566) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5550..5566) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 5997..6788 /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 6881..7469 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7370..7389 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7623..7640 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg 121 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt 181 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt 241 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa 301 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct 361 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta 421 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga 481 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa 541 aagttggcac catgggagcc atcgggctcc tgtggctgct gccgctgctg ctttccacgg 601 cagctgtggg ctccgggatg gggaccggcc agcgcgcggg ctccccagct gcggggtcgc 661 cgctgcagcc ccgggagcca ctcagctact cgcgcctgca gaggaagagt ctggcagttg 721 acttcgtggt gccctcgctc ttccgtgtct acgcccggga cctactgctg ccaccatcct 781 cctcggagct gaaggctggc aggcccgagg cccgcggctc gctagctctg gactgcgccc 841 cgctgctcag gttgctgggg ccggcgccgg gggtctcctg gaccgccggt tcaccagccc 901 cggcagaggc ccggacgctg tccagggtgc tgaagggcgg ctccgtgcgc aagctccggc 961 gtgccaagca gttggtgctg gagctgggcg aggaggcgat cttggagggt tgcgtcgggc 1021 cccccgggga ggcggctgtg gggctgctcc agttcaatct cagcgagctg ttcagttggt 1081 ggattcgcca aggcgaaggg cgactgagga tccgcctgat gcccgagaag aaggcgtcgg 1141 aagtgggcag agagggaagg ctgtccgcgg caattcgcgc ctcccagccc cgccttctct 1201 tccagatctt cgggactggt catagctcct tggaatcacc aacaaacatg ccatctcctt 1261 ctcctgatta ttttacatgg aatctcacct ggataatgaa agactccttc cctttcctgt 1321 ctcatcgcag ccgatatggt ctggagtgca gctttgactt cccctgtgag ctggagtatt 1381 cccctccact gcatgacctc aggaaccaga gctggtcctg gcgccgcatc ccctccgagg 1441 aggcctccca gatggacttg ctggatgggc ctggggcaga gcgttctaag gagatgccca 1501 gaggctcctt tctccttctc aacacctcag ctgactccaa gcacaccatc ctgagtccgt 1561 ggatgaggag cagcagtgag cactgcacac tggccgtctc ggtgcacagg cacctgcagc 1621 cctctggaag gtacattgcc cagctgctgc cccacaacga ggctgcaaga gagatcctcc 1681 tgatgcccac tccagggaag catggttgga cagtgctcca gggaagaatc gggcgtccag 1741 acaacccatt tcgagtggcc ctggaataca tctccagtgg aaaccgcagc ttgtctgcag 1801 tggacttctt tgccctgaag aactgcagtg aaggaacatc cccaggctcc aagatggccc 1861 tgcagagctc cttcacttgt tggaatggga cagtcctcca gcttgggcag gcctgtgact 1921 tccaccagga ctgtgcccag ggagaagatg agagccagat gtgccggaaa ctgcctgtgg 1981 gtttttactg caactttgaa gatggcttct gtggctggac ccaaggcaca ctgtcacccc 2041 acactcctca gtggcaggtc aggaccctaa aggatgcccg gttccaggac caccaagacc 2101 atgctctatt gctcagtacc actgatgtcc ccgcttctga aagtgctaca gtgaccagtg 2161 ctacgtttcc tgcaccgatc aagagctctc catgtgagct ccgaatgtcc tggctcattc 2221 gtggagtctt gaggggaaac gtgtccttgg tgctagtgga gaacaaaacc gggaaggagc 2281 aaggcaggat ggtctggcat gtcgccgcct atgaaggctt gagcctgtgg cagtggatgg 2341 tgttgcctct cctcgatgtg tctgacaggt tctggctgca gatggtcgca tggtggggac 2401 aaggatccag agccatcgtg gcttttgaca atatctccat cagcctggac tgctacctca 2461 ccattagcgg agaggacaag atcctgcaga atacagcacc caaatcaaga aacctgtttg 2521 agagaaaccc aaacaaggag ctgaaacccg gggaaaattc accaagacag acccccatct 2581 ttgaccctac agttcattgg ctgttcacca catgtggggc cagcgggccc catggcccca 2641 cccaggcaca gtgcaacaac gcctaccaga actccaacct gagcgtggag gtggggagcg 2701 agggccccct gaaaggcatc cagatctgga aggtgccagc caccgacacc tacagcatct 2761 cgggctacgg agctgctggc gggaaaggcg ggaagaacac catgatgcgg tcccacggcg 2821 tgtctgtgct gggcatcttc aacctggaga aggatgacat gctgtacatc ctggttgggc 2881 agcagggaga ggacgcctgc cccagtacaa accagttaat ccagaaagtc tgcattggag 2941 agaacaatgt gatagaagaa gaaatccgtg tgaacagaag cgtgcatgag tgggcaggag 3001 gcggaggagg agggggtgga gccacctacg tatttaagat gaaggatgga gtgccggtgc 3061 ccctgatcat tgcagccgga ggtggcggca gggcctacgg ggccaagaca gacacgttcc 3121 acccagagag actggagaat aactcctcgg ttctagggct aaacggcaat tccggagccg 3181 caggtggtgg aggtggctgg aatgataaca cttccttgct ctgggccgga aaatctttgc 3241 aggagggtgc caccggagga cattcctgcc cccaggccat gaagaagtgg gggtgggaga 3301 caagaggggg tttcggaggg ggtggagggg ggtgctcctc aggtggagga ggcggaggat 3361 atataggcgg caatgcagcc tcaaacaatg accccgaaat ggatggggaa gatggggttt 3421 ccttcatcag tccactgggc atcctgtaca ccccagcttt aaaagtgatg gaaggccacg 3481 gggaagtgaa tattaagcat tatctaaact gcagtcactg tgaggtagac gaatgtcaca 3541 tggaccctga aagccacaag gtcatctgct tctgtgacca cgggacggtg ctggctgagg 3601 atggcgtctc ctgcattgtg tcacccaccc cggagccaca cctgccactc tcgctgatcc 3661 tctctgtggt gacctctgcc ctcgtggccg ccctggtcct ggctttctcc ggcatcatga 3721 ttgtgtaccg ccggaagcac caggagctgc aagccatgca gatggagctg cagagccctg 3781 agtacaagct gagcaagctc cgcacctcga ccatcatgac cgactacaac cccaactact 3841 gctttgctgg caagacctcc tccatcagtg acctgaagga ggtgccgcgg aaaaacatca 3901 ccctcattcg gggtctgggc catggcgcct ttggggaggt gtatgaaggc caggtgtccg 3961 gaatgcccaa cgacccaagc cccctgcaag tggctgtgaa gacgctgcct gaagtgtgct 4021 ctgaacagga cgaactggat ttcctcatgg aagccctgat catcagcaaa ttcaaccacc 4081 agaacattgt tcgctgcatt ggggtgagcc tgcaatccct gccccggttc atcctgctgg 4141 agctcatggc ggggggagac ctcaagtcct tcctccgaga gacccgccct cgcccgagcc 4201 agccctcctc cctggccatg ctggaccttc tgcacgtggc tcgggacatt gcctgtggct 4261 gtcagtattt ggaggaaaac cacttcatcc accgagacat tgctgccaga aactgcctct 4321 tgacctgtcc aggccctgga agagtggcca agattggaga cttcgggatg gcccgagaca 4381 tctacagggc gagctactat agaaagggag gctgtgccat gctgccagtt aagtggatgc 4441 ccccagaggc cttcatggaa ggaatattca cttctaaaac agacacatgg tcctttggag 4501 tgctgctatg ggaaatcttt tctcttggat atatgccata ccccagcaaa agcaaccagg 4561 aagttctgga gtttgtcacc agtggaggcc ggatggaccc acccaagaac tgccctgggc 4621 ctgtataccg gataatgact cagtgctggc aacatcagcc tgaagacagg cccaactttg 4681 ccatcatttt ggagaggatt gaatactgca cccaggaccc ggatgtaatc aacaccgctt 4741 tgccgataga atatggtcca cttgtggaag aggaagagaa agtgcctgtg aggcccaagg 4801 accctgaggg ggttcctcct ctcctggtct ctcaacaggc aaaacgggag gaggagcgca 4861 gcccagctgc cccaccacct ctgcctacca cctcctctgg caaggctgca aagaaaccca 4921 cagctgcaga ggtctctgtt cgagtcccta gagggccggc cgtggaaggg ggacacgtga 4981 atatggcatt ctctcagtcc aaccctcctt cggagttgca caaggtccac ggatccagaa 5041 acaagcccac cagcttgtgg aacccaacgt acggctcctg gtttacagag aaacccacca 5101 aaaagaataa tcctatagca aagaaggagc cacacgacag gggtaacctg gggctggagg 5161 gaagctgtac tgtcccacct aacgttgcaa ctgggagact tccgggggcc tcactgctcc 5221 tagagccctc ttcgctgact gccaatatga aggaggtacc tctgttcagg ctacgtcact 5281 tcccttgtgg gaatgtcaat tacggctacc agcaacaggg cttgccctta gaagccgcta 5341 ctgcccctgg agctggtcat tacgaggata ccattctgaa aagcaagaat agcatgaacc 5401 agcctgggcc cccaactttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa 5461 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg 5521 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg 5581 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgcctcctct 5641 agaccagcca ggacagaaat gcctcgactt cgctgctacc caaggttgcc gggtgacgca 5701 caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg gttcgtaagc 5761 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg 5821 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt ggggtacagt 5881 ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat 5941 ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta aacattatga 6001 gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc 6061 gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc 6121 tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa 6181 cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga 6241 ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc 6301 cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct 6361 tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata 6421 gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc 6481 tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg 6541 atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa 6601 tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc 6661 ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc 6721 gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg 6781 gcaaataacc ctcgagccac ccatgaccaa aatcccttaa cgtgagttac gcgtcgttcc 6841 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc 6901 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg 6961 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa 7021 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc 7081 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt 7141 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa 7201 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc 7261 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc 7321 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct 7381 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat 7441 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc 7501 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg 7561 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc 7621 gcagcgagtc agtgagcgag gaagcggaag //