LOCUS Exported 3933 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDONR223-CKB SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3933) 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 3933) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3933 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 1..100 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(1244..1343) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1360..1379) /label=T7 /note="T7 promoter, forward primer" promoter complement(1361..1379) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1384..1400) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1384..1400) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 1831..2622 /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 2715..3303 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3204..3223 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3457..3474 /label=L4440 /note="L4440 vector, forward primer" terminator 3633..3660 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 3752..3838 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 3887..3909 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 3901..3918 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 3902..3918 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa 61 tgctttttta taatgccaac tttgtacaaa aaagttggca tgcccttctc caacagccac 121 aacgcactga agctgcgctt cccggccgag gacgagttcc ccgacctgag cgcccacaac 181 aaccacatgg ccaaggtgct gacccccgag ctgtacgcgg agctgcgcgc caagagcacg 241 ccgagcggct tcacgctgga cgacgtcatc cagacaggcg tggacaaccc gggccacccg 301 tacatcatga ccgtgggctg cgtggcgggc gacgaggagt cctacgaagt gttcaaggat 361 ctcttcgacc ccatcatcga ggaccggcac ggcggctaca agcccagcga tgagcacaag 421 accgacctca accccgacaa cctgcagggc ggcgacgacc tggaccccaa ctacgtgctg 481 agctcgcggg tgcgcacggg ccgcagcatc cgtggcttct gcctcccccc gcactgcagc 541 cgcggggagc gccgagccat cgagaagctc gcggtggaag ccctgtccag cctggacggc 601 gacctggcgg gccgatacta cgcgctcaag agcatgacgg aggcggagca gcagcagctc 661 atcgacgacc acttcctctt cgacaagccc gtgtcgcccc tgctgctggc ctcgggcatg 721 gcccgcgact ggcccgacgc ccgcggtatc tggcacaatg acaataagac cttcctggtg 781 tgggtcaacg aggaggacca cctgcgggtc atctccatgc agaagggggg caacatgaag 841 gaggtgttca cccgcttctg caccggcctc acccagattg aaactctctt caagtctaag 901 gactatgagt tcatgtggaa ccctcacctg ggctacatcc tcacctgccc atccaacctg 961 ggcaccgggc tgcgggcagg tgtgcatatc aagctgccca acctgggcaa gcatgagaag 1021 ttctcggagg tgcttaagcg gctgcgactt cagaagcgag gcacaggcgg tgtggacacg 1081 gctgcggtgg gcggggtctt cgacgtctcc aacgctgacc gcctgggctt ctcagaggtg 1141 gagctggtgc agatggtggt ggacggagtg aagctgctca tcgagatgga acagcggctg 1201 gagcagggcc aggccatcga cgacctcatg cctgcccaga aatgcccaac tttcttgtac 1261 aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc 1321 agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac 1381 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt 1441 gcacaagata aaaatatatc atcatgcctc ctctagacca gccaggacag aaatgcctcg 1501 acttcgctgc tacccaaggt tgccgggtga cgcacaccgt ggaaacggat gaaggcacga 1561 acccagtgga cataagcctg ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca 1621 cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc 1681 atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa 1741 gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag 1801 ggcagtcgcc ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc cgaagtatcg 1861 actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc 1921 gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg 1981 ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt 2041 ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt 2101 gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga 2161 gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat 2221 ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg 2281 gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta 2341 acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg 2401 tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac 2461 tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct 2521 tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc 2581 cactacgtga aaggcgagat caccaaggta gtcggcaaat aaccctcgag ccacccatga 2641 ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag 2701 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa 2761 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg 2821 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag 2881 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg 2941 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga 3001 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc 3061 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc 3121 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga 3181 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt 3241 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg 3301 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac 3361 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga 3421 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg 3481 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc 3541 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt 3601 accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct 3661 gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt 3721 gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg 3781 acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat 3841 gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt 3901 tgtaaaacga cggccagtct taagctcggg ccc //