LOCUS Exported 4185 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDONR223-GCK SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4185) 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 4185) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4185 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..17 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 33..132 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(1528..1627) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1644..1663) /label=T7 /note="T7 promoter, forward primer" promoter complement(1645..1663) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1668..1684) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1668..1684) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 2115..2906 /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 2999..3587 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3488..3507 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3741..3758 /label=L4440 /note="L4440 vector, forward primer" terminator 3917..3944 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 4036..4122 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 4171..8 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 4185..17 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" ORIGIN 1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag 61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca 121 aaaaagttgg catgctggac gacagagcca ggatggaggc cgccaagaag gagaaggtag 181 agcagatcct ggcagagttc cagctgcagg aggaggacct gaagaaggtg atgagacgga 241 tgcagaagga gatggaccgc ggcctgaggc tggagaccca tgaagaggcc agtgtgaaga 301 tgctgcccac ctacgtgcgc tccaccccag aaggctcaga agtcggggac ttcctctccc 361 tggacctggg tggcactaac ttcagggtga tgctggtgaa ggtgggagaa ggtgaggagg 421 ggcagtggag cgtgaagacc aaacaccaga tgtactccat ccccgaggac gccatgaccg 481 gcactgctga gatgctcttc gactacatct ctgagtgcat ctccgacttc ctggacaagc 541 atcagatgaa acacaagaag ctgcccctgg gcttcacctt ctcctttcct gtgaggcacg 601 aagacatcga taagggcatc cttctcaact ggaccaaggg cttcaaggcc tcaggagcag 661 aagggaacaa tgtcgtgggg cttctgcgag acgctatcaa acggagaggg gactttgaaa 721 tggatgtggt ggcaatggtg aatgacacgg tggccacgat gatctcctgc tactacgaag 781 accatcagtg cgaggtcggc atgatcgtgg gcacgggctg caatgcctgc tacatggagg 841 agatgcagaa tgtggagctg gtggaggggg acgagggccg catgtgcgtc aataccgagt 901 ggggcgcctt cggggactcc ggcgagctgg acgagttcct gctggagtat gaccgcctgg 961 tggacgagag ctctgcaaac cccggtcagc agctgtatga gaagctcata ggtggcaagt 1021 acatgggcga gctggtgcgg cttgtgctgc tcaggctcgt ggacgaaaac ctgctcttcc 1081 acggggaggc ctccgagcag ctgcgcacac gcggagcctt cgagacgcgc ttcgtgtcgc 1141 aggtggagag cgacacgggc gaccgcaagc agatctacaa catcctgagc acgctggggc 1201 tgcgaccctc gaccaccgac tgcgacatcg tgcgccgcgc ctgcgagagc gtgtctacgc 1261 gcgctgcgca catgtgctcg gcggggctgg cgggcgtcat caaccgcatg cgcgagagcc 1321 gcagcgagga cgtaatgcgc atcactgtgg gcgtggatgg ctccgtgtac aagctgcacc 1381 ccagcttcaa ggagcggttc catgccagcg tgcgcaggct gacgcccagc tgcgagatca 1441 ccttcatcga gtcggaggag ggcagtggcc ggggcgcggc cctggtctcg gcggtggcct 1501 gtaagaaggc ctgtatgctg ggccagtgcc caactttctt gtacaaagtt ggcattataa 1561 gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca 1621 ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt 1681 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata 1741 tatcatcatg cctcctctag accagccagg acagaaatgc ctcgacttcg ctgctaccca 1801 aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag tggacataag 1861 cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa 1921 ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact 1981 gtttttttgg ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg 2041 gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa 2101 caaagttaaa cattatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg 2161 tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct 2221 ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg 2281 taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt 2341 cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca 2401 tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg 2461 acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga 2521 caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc 2581 cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc 2641 cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca 2701 gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc 2761 tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag 2821 aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg 2881 agatcaccaa ggtagtcggc aaataaccct cgagccaccc atgaccaaaa tcccttaacg 2941 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt 3001 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag 3061 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca 3121 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca 3181 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg 3241 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg 3301 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct 3361 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga 3421 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc 3481 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg 3541 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg 3601 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt 3661 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc 3721 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac 3781 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc 3841 ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag 3901 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct 3961 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa 4021 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa 4081 acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta 4141 ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgtt //