LOCUS       sequence-437686-        4842 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 4842)
  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 4842)
  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-NEK5
FEATURES             Location/Qualifiers
     source          1..4842
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(16..102)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     151..173
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     165..182
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    198..297
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2350..2449)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2466..2485)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2467..2485)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2490..2506)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2490..2506)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2937..3728
                     /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      3821..4409
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4310..4329
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4563..4580
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4739..4766)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
       61 acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta
      121 ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtcttaagct cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc
      241 aacaaattga tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa gttggcacca
      301 tggataagta cgatgtgatt aaggccatcg ggcaaggtgc cttcgggaaa gcatacttag
      361 ctaaagggaa atcagatagc aagcactgtg tcataaaaga gatcaatttt gaaaagatgc
      421 ccatacaaga aaaagaagct tcaaagaaag aagtgattct tctggaaaag atgaaacatc
      481 ccaacattgt agccttcttc aattcatttc aagagaatgg caggctgttt attgtaatgg
      541 aatattgtga tggaggggat ctcatgaaaa ggatcaatag acaacggggt gtgttattta
      601 gtgaagatca gatcctcggt tggtttgtac agatttctct aggactaaaa catattcatg
      661 acaggaagat attacacagg gacataaaag ctcagaacat ttttcttagc aagaacggaa
      721 tggtggcaaa gcttggggac tttggtatag caagagtcct gaataattcc atggaacttg
      781 ctcgaacttg tattggaaca ccttactacc tgtccccaga gatctgtcag aataaaccct
      841 acaacaataa aacggatatt tggtctcttg gctgtgtctt atatgagctc tgcacactta
      901 aacatccttt tgagggtaac aacttacagc agctggttct gaagatttgt caagcacatt
      961 ttgccccaat atctccgggg ttttctcgtg agctccattc cttgatatct cagctctttc
     1021 aagtatctcc tcgagaccga ccatccataa attccatttt gaaaaggccc tttttagaga
     1081 atcttattcc caaatatttg actcctgagg tcattcagga agaattcagt cacatgctta
     1141 tatgcagagc aggagcgcca gcttctcgac atgctgggaa ggtggtccag aagtgtaaaa
     1201 tacaaaaagt gagattccag ggaaagtgcc caccaagatc aaggatatct gtgccaatta
     1261 aaaggaatgc tatattgcat agaaatgaat ggagaccacc agctggagcc cagaaggcca
     1321 gatctataaa aatgatagaa agacccaaaa ttgctgctgt ctgtggacat tatgattatt
     1381 attatgctca acttgatatg ctgaggagga gagcccacaa accaagttat caccctattc
     1441 ctcaagaaaa tactggagtt gaggattacg gtcaggaaac gaggcatggt ccatccccaa
     1501 gtcaatggcc tgctgagtac cttcagagaa aatttgaagc tcaacaatat aagttgaaag
     1561 tggagaagca attgggtctt cgtccatctt ctgccgagcc aaattacaac cagagacaag
     1621 agctaagaag taatggagaa gagcctagat tccaggagct gccatttagg aaaaacgaaa
     1681 tgaaggaaca ggaatattgg aagcagttag aggaaatacg ccaacagtac cacaatgaca
     1741 tgaaagaaat tagaaagaag atggggagag aaccagagga gaactcaaaa ataagtcata
     1801 aaacctattt ggtgaagaag agtaacctgc ctgtccatca agatgcatct gagggagaag
     1861 cacctgtgca ggacattgaa aaagacttga aacaaatgag gcttcagaac acaaaggaaa
     1921 gtaaaaatcc agaacagaaa tataaagcta agaagggggt aaaatttgaa attaatttag
     1981 acaaatgtat ttctgatgaa aacatcctcc aagaggaaga ggcaatggat ataccaaatg
     2041 aaactttgac ctttgaggat ggcatgaagt ttaaggaata tgaatgtgta aaggagcatg
     2101 gagattatac agacaaagca tttgaaaaac ttcactgccc agaagcaggg ttttccacgc
     2161 agactgtagc tgctgtggga aacaggaggc agtgggatgg aggagcgcct cagactctgc
     2221 tgcagatgat ggcagtggcc gacatcacct ccacctgccc cacggggcct gacagtgagt
     2281 ctgtgcttag cgtcagtcgt caggaaggga agaccaagga cccgtacagc ccagtgctca
     2341 tcctgatgtt gccaactttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa
     2401 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
     2461 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
     2521 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgcctcctct
     2581 agaccagcca ggacagaaat gcctcgactt cgctgctacc caaggttgcc gggtgacgca
     2641 caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg gttcgtaagc
     2701 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
     2761 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt ggggtacagt
     2821 ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat
     2881 ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta aacattatga
     2941 gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc
     3001 gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc
     3061 tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa
     3121 cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga
     3181 ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc
     3241 cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct
     3301 tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata
     3361 gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc
     3421 tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg
     3481 atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa
     3541 tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc
     3601 ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc
     3661 gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg
     3721 gcaaataacc ctcgagccac ccatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
     3781 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3841 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3901 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3961 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4021 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4081 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4141 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4201 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4261 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4321 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4381 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4441 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     4501 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     4561 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     4621 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     4681 gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa
     4741 aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg
     4801 tcctgcccgc caccctccgg gccgttgctt cacaacgttc aa
//
