LOCUS Exported 6972 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDONR223-LRPPRC SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6972) 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 6972) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6972 /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(4732..4831) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(4848..4867) /label=T7 /note="T7 promoter, forward primer" promoter complement(4849..4867) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4872..4888) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4872..4888) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 5319..6110 /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 6203..6791 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6692..6711 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6945..6962 /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 catggcagcc ctgctgagat ccgcgcgttg gttgctgcgt gccggggcgg 601 ccccgcgcct cccgctctcc ctgcgcctcc tccctggcgg cccgggccgg ctgcatgccg 661 cctcctatct gcccgccgct cgcgccgggc ccgtggccgg aggactactg agcccagcca 721 ggctgtatgc cattgctgcc aaagaaaaag atattcaaga ggagtccact ttttcttcta 781 ggaagatttc caatcaattt gattgggctc taatgagact agatctttct gttcgaagaa 841 ctggccgcat tccaaagaag cttctacaaa aagtttttaa tgatacctgc cgctcaggtg 901 gcctaggtgg tagtcatgcc ttgcttctac tacgtagttg tggttctctc ttgcctgaac 961 taaagcttga agagagaaca gaatttgctc ataggatatg ggacacactt cagaaattag 1021 gtgctgtgta tgatgtgagt cactataatg ctttacttaa agtctatctt caaaatgaat 1081 ataaattctc accaactgat ttcctggcaa aaatggagga agcaaacatt caaccaaatc 1141 gagtgacata ccagagattg attgcttctt attgtaatgt aggagatatt gaaggtgcca 1201 gcaagattct tggatttatg aaaactaagg atctcccagt tacagaggca gtattcagtg 1261 cccttgtgac agggcatgcc agagctggtg atatggagaa tgcagaaaac attctcacag 1321 tgatgagaga tgccggaatt gagcctggtc cagacacata cctcgcatta ttgaatgcat 1381 atgctgagaa gggcgacatt gaccatgtta agcagactct ggagaaggtg gagaagtccg 1441 agcttcacct tatggaccgt gatttactgc aaattatttt tagcttcagt aaagctgggt 1501 atcctcagta tgtctcagaa attttggaaa aagttacatg tgaaagaaga tatattccag 1561 atgcaatgaa cctcatttta cttttagtca ctgaaaaatt ggaagatgta gcgttgcaaa 1621 ttttactagc atgccccgta tcaaaggaag atggcccaag tgtctttggc agtttctttt 1681 tacaacactg tgtgactatg aatacgcctg tggagaagct aacagactac tgtaagaagt 1741 taaaggaagt ccagatgcac tcctttcctc tgcagttcac cctccattgt gctttactcg 1801 ccaataaaac tgatttggca aaagccttaa tgaaggctgt gaaggaggaa ggttttccta 1861 tcagacctca ctatttctgg ccattgctag ttggacgtcg gaaggaaaaa aatgttcaag 1921 gtataattga aatcctcaaa ggaatgcaag aattgggagt acatcctgat caggaaacat 1981 atacagatta tgtgattcca tgctttgata gtgtaaactc agcacgagcc attttgcagg 2041 aaaatggatg tctgtctgat agtgatatgt tttctcaagc tggattgaga agtgaagcag 2101 caaatgggaa cttagacttt gtattatcat ttttgaaatc aaatacattg cccatctcgc 2161 tgcagtctat aagaagtagc ctactgctag gcttcaggag gtctatgaat ataaatcttt 2221 ggagcgagat aacagaattg ttgtacaagg atggacgtta ttgccaggag cctcgaggac 2281 cgacggctgt tggctatttt ctttataact tgattgacag catgagtgac tcagaggtac 2341 aggccaagga ggagcatttg agacaatact tccatcagct ggagaagatg aatgtaaaaa 2401 ttcctgaaaa tatctacaga ggcattcgta atctcctgga aagctaccat gttcctgaat 2461 tgattaagga tgctcacttg ttggttgaga gtaagaattt agactttcaa aaaactgtgc 2521 aacttacatc atctgaattg gagtccacac ttgaaacact aaaagctgaa aatcaaccta 2581 taagagatgt cctaaagcaa ctcatattag tgctttgttc agaagagaat atgcaaaaag 2641 cccttgaatt gaaagcaaaa tatgaatccg acatggttac tggtggctat gcagctttaa 2701 taaatttatg ctgtcgacat gataaagtag aagatgcctt gaacttgaaa gaagaatttg 2761 accgcttaga ttcatctgct gtccttgaca ccggcaagta tgtaggcctt gtaagagtat 2821 tggcaaagca tggcaagctc caagatgcta ttaacattct gaaggagatg aaagagaagg 2881 atgttcttat caaagataca acagccttgt cctttttcca catgctaaat ggcgcagctt 2941 taagaggtga aattgaaaca gtaaaacagt tgcatgaagc catcgtgact ctagggttag 3001 cagaaccatc caccaacata agtttcccat tggtcactgt acacttggaa aagggcgacc 3061 tatctactgc tcttgaggtc gccattgact gctatgaaaa gtataaagta ttaccaagga 3121 ttcatgatgt cttgtgtaaa ctggtagaga aaggcgagac tgatctaatt cagaaagcaa 3181 tggactttgt gagccaagaa caaggtgaaa tggtgatgct ctatgatctc ttctttgcct 3241 tcctacaaac aggaaattac aaagaggcca agaagatcat tgagactcca gggattagag 3301 ctcgatctgc aaggcttcag tggttttgtg acagatgtgt tgcaaataat caggttgaaa 3361 ctctggaaaa attagtggag ctgacacaga agctatttga atgtgataga gaccagatgt 3421 actacaatct gctaaaactg tataaaataa acggtgactg gcaaagagct gatgcagtct 3481 ggaataaaat ccaagaagaa aatgttattc ctcgtgaaaa gacattaaga ttattagcag 3541 aaatccttag agagggtaac caggaagttc cgtttgacgt acctgagttg tggtatgaag 3601 atgaaaaaca ttccctgaat tcttcgtcag cctcaaccac agaacctgat ttccagaaag 3661 atatattgat tgcctgccga ttgaaccaaa aaaaaggggc atatgatatt ttcctgaatg 3721 caaaagagca aaacattgtg tttaatgctg aaacctacag caatctcatt aaattactga 3781 tgtcagaaga ttattttaca caagcaatgg aagtgaaagc attcgcggag acccacatca 3841 agggcttcac actgaacgat gctgccaaca gccgcctcat cataacgcaa gttaggcggg 3901 attatttgaa agaggctgtg acaacactga aaacagtatt ggatcagcag cagacccctt 3961 ctaggttagc agtgacccgt gtcatccagg cattggccat gaagggtgat gttgaaaaca 4021 tagaagtagt tcagaagatg ttaaatggac tcgaagactc cattggactt tcaaaaatgg 4081 ttttcatcaa taacattgct ttggctcaaa taaagaataa taacatagat gccgcaatag 4141 aaaacattga aaatatgctt acttcagaga ataaagtcat tgaaccccaa tacttcggct 4201 tggcatactt attcagaaaa gtaatagagg agcagttgga accagcagtt gaaaagataa 4261 gcatcatggc ggagagattg gccaatcagt ttgcaattta taaacctgtc actgattttt 4321 tccttcaact tgtggatgca ggcaaggtgg atgatgccag agctctccta cagagatgtg 4381 gtgcaattgc tgaacaaacc ccgattttgt tgttgttcct ccttaggaat tctaggaaac 4441 aaggaaaggc atcaactgtg aaatctgtgt tagaattgat tcctgaatta aatgaaaagg 4501 aagaagcata caattccctc atgaaaagct atgtctcaga gaaagatgtc acatctgcta 4561 aagcactgta tgaacatttg actgcaaaga atacaaaatt ggatgatctg tttctaaagc 4621 gttacgcatc tttgctgaag tatgctggag agcctgtccc tttcattgaa ccccctgaaa 4681 gctttgaatt ttatgcacag cagctaagaa aattgaggga aaactcttct ttgccaactt 4741 tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg 4801 tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt 4861 cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat 4921 gttacattgc acaagataaa aatatatcat catgcctcct ctagaccagc caggacagaa 4981 atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg aaacggatga 5041 aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta 5101 tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg 5161 cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca 5221 agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta 5281 cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg gtgatcgccg 5341 aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt 5401 tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata 5461 ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca 5521 acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag 5581 tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc 5641 aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg 5701 acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc 5761 cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg 5821 aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc 5881 ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg 5941 ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta 6001 gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag 6061 aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc 6121 acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg 6181 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 6241 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 6301 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt 6361 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 6421 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 6481 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 6541 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 6601 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 6661 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 6721 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 6781 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 6841 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 6901 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 6961 aggaagcgga ag //