LOCUS       sequence-402568-        5730 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5730)
  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 Dec 16;468(7326):968-72. doi: 10.1038/nature09627
  PUBMED    21107320
REFERENCE   2  (bases 1 to 5730)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nature"; date: "2010-12-16- 16"; volume: "468"; issue: "7326"; 
            pages: "968-72"
COMMENT     Sequence Label: pDONR223-NPR2
FEATURES             Location/Qualifiers
     source          1..5730
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     38..60
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     52..69
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     53..69
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    85..184
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(1506..1523)
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="GSSKSK"
     protein_bind    complement(3125..3224)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3241..3260)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3242..3260)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3265..3281)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3265..3281)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3712..4503
                     /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      4596..5184
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5085..5104
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5338..5355
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5514..5541)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5633..5719)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg ttgtaaaacg
       61 acggccagtc ttaagctcgg gccccaaata atgattttat tttgactgat agtgacctgt
      121 tcgttgcaac aaattgatga gcaatgcttt tttataatgc caactttgta caaaaaagtt
      181 ggcaccatgg cgctgccatc acttctgctg ttggtggcag ccctggcagg tggggtgcgt
      241 cctcccgggg cgcggaacct gacgctggcg gtggtgctgc cagaacacaa cctgagctat
      301 gcctgggcct ggccacgggt gggacccgct gtggcactag ctgtggaggc tctgggccgg
      361 gcactgcccg tggacctgcg gtttgtcagc tccgaactgg aaggcgcctg ctctgagtac
      421 ctggcaccgc tgagcgctgt ggacctcaag ctgtaccatg accccgacct gctgttaggt
      481 cccggttgcg tgtaccctgc tgcctctgtg gcccgctttg cctcccactg gcgccttccc
      541 ctgctgactg cgggtgctgt ggcctctggt ttttcggcta agaatgacca ttatcgtacc
      601 ctggttcgca ctggcccctc tgctcccaag ctgggtgagt ttgtggtgac actacacggg
      661 cacttcaatt ggactgcccg tgctgccttg ctgtacctgg atgctcgcac agatgaccgg
      721 cctcactact tcaccatcga gggcgtcttt gaggccctgc agggcagcaa cctcagtgtg
      781 cagcaccagg tgtatgcccg agagccaggg ggccccgagc aggccaccca cttcatccgg
      841 gccaacgggc gcattgtgta tatctgcggc cctctggaga tgctgcatga gatcctgctt
      901 caggcccaga gggagaatct gaccaatggg gattatgtct tcttttacct ggatgtcttt
      961 ggggagagtc tccgtgcagg ccccacacgt gctacaggcc ggccctggca ggacaatcgc
     1021 acccgggaac aggcccaggc cctcagagag gcctttcaga ctgtattggt gatcacgtac
     1081 cgagaacccc caaatcctga gtatcaggaa ttccagaatc gtctgctgat aagagcccgg
     1141 gaagactttg gtgtggagct gggcccttcc ctgatgaacc tcatcgctgg ctgcttctat
     1201 gatgggatcc tgctatatgc tgaagtcctg aatgagacaa tacaggaagg aggcacccgg
     1261 gaggatggac ttcgaattgt ggaaaagatg cagggacgaa gatatcacgg tgtaactggg
     1321 ctggttgtca tggacaagaa caatgaccga gagactgact ttgtcctctg ggccatggga
     1381 gacctggatt ctggggactt tcagcctgca gcccactact cgggagctga gaagcagatt
     1441 tggtggacgg gacggcctat tccctgggtg aagggggctc ctccctcgga caatcccccc
     1501 tgtgcctttg acttggacga cccatcctgt gataaaactc cactttcaac cctggcaatt
     1561 gtggctctgg gcacaggaat caccttcatc atgtttggtg tttccagctt cctaattttc
     1621 cgaaagctga tgctggagaa ggagctggct agcatgttgt ggcgtattcg ctgggaagaa
     1681 ctgcagtttg gcaactcaga gcgttatcac aaaggtgcag gcagtcgcct cacactgtcg
     1741 ctgcggggat ccagttacgg ctcgctcatg acagcccatg ggaaatacca gatctttgcc
     1801 aacaccggtc acttcaaggg aaatgttgtc gccatcaaac atgtgaataa gaagcgcatt
     1861 gagctgaccc ggcaggttct gtttgaactc aaacatatga gagatgttca gttcaaccat
     1921 ctcactcgct tcattggcgc ctgcatagac cctcccaaca tttgcattgt cactgaatac
     1981 tgtcctcgtg ggagtttaca gggcatggcc tttctccaca acagcattat ttcatcgcat
     2041 gggagtctca agtcctccaa ctgtgtggtg gatagtcgtt ttgtgctcaa aatcacagac
     2101 tatggcctgg ccagcttccg atcaactgct gaacctgatg acagccatgc cctctatgcc
     2161 aagaagctgt ggactgcccc agaactgctc agtgggaacc ccttgccaac cacaggcatg
     2221 cagaaggctg acgtctatag ctttgggatc atcctgcagg agatagcact tcgcagtggt
     2281 cctttctact tggagggcct ggacctcagc cccaaagaga ttgtccagaa ggtacgaaat
     2341 ggtcagcggc catatttccg gccaagcatt gaccggaccc aactgaatga agagctagtt
     2401 ttgctgatgg agcgatgttg ggctcaggac ccagctgagc ggccagactt tggacagatt
     2461 aagggcttca ttcggcgctt taacaaggag ggtggcacca gcatattgga caacctcctg
     2521 ctgcgcatgg aacagtatgc caataacttg gagaagctgg tggaggaacg cacacaggcc
     2581 tatctggagg aaaaacgcaa ggctgaagct ctgctctacc aaatcctacc ccattcagtg
     2641 gcagagcagt taaaacgggg agagactgta caggctgagg cctttgacag tgttaccatc
     2701 tacttcagtg acattgttgg cttcacagca ttgtcagcag agagcacccc catgcaggta
     2761 gtgacacttc ttaatgacct gtatacctgc tttgatgcca taattgacaa ctttgatgtc
     2821 tacaagtctc cacctttccc agactctccc aacctgtttc ttctcaaagg gccagtctgt
     2881 gctggggttg ttggcctgaa gatgccccgt tattgtcttt ttggagacac agtgaacact
     2941 gcttctcgaa tggagtctaa tggtcaagcg ctgaagatcc atgtctcctc taccaccaag
     3001 gatgccctag atgagctagg atgcttccag ctagagcttc ggggggatgt ggaaatgaag
     3061 ggaaaaggaa agatgcgaac atactggctc ttaggagagc ggaaaggacc tcctggactc
     3121 ctgttgccaa ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
     3181 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc
     3241 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
     3301 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgcct cctctagacc
     3361 agccaggaca gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg acgcacaccg
     3421 tggaaacgga tgaaggcacg aacccagtgg acataagcct gttcggttcg taagctgtaa
     3481 tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt
     3541 aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg
     3601 cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc
     3661 agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat tatgagggaa
     3721 gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat
     3781 ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag
     3841 ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg
     3901 cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc
     3961 cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct
     4021 aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag
     4081 ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt
     4141 gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt
     4201 gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag
     4261 cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg
     4321 ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc
     4381 atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca
     4441 gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa
     4501 taaccctcga gccacccatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     4561 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4621 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4681 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4741 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4801 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4861 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4921 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4981 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5041 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5101 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5161 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5221 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     5281 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     5341 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     5401 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     5461 cgcaacgcaa ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc
     5521 catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc gggcgtcctg
     5581 cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc
     5641 tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag tcttccgact
     5701 gagcctttcg ttttatttga tgcctggcag
//
