LOCUS       sequence-457343-        3699 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 3699)
  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 3699)
  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-NEK7
FEATURES             Location/Qualifiers
     source          1..3699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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(1010..1109)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1126..1145)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1127..1145)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1150..1166)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1150..1166)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1597..2388
                     /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      2481..3069
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2970..2989
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3223..3240
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3399..3426)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3518..3604)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3653..3675
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3667..3684
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3668..3684
                     /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 ccatggatga gcaatcacaa
      121 ggaatgcaag ggccacctgt tcctcagttc caaccacaga aggccttacg accggatatg
      181 ggctataata cattagccaa ctttcgaata gaaaagaaaa ttggtcgcgg acaatttagt
      241 gaagtttata gagcagcctg tctcttggat ggagtaccag tagctttaaa aaaagtgcag
      301 atatttgatt taatggatgc caaagcacgt gctgattgca tcaaagaaat agatcttctt
      361 aagcaactca accatccaaa tgtaataaaa tattatgcat cattcattga agataatgaa
      421 ctaaacatag ttttggaact agcagatgct ggcgacctat ccagaatgat caagcatttt
      481 aagaagcaaa agaggctaat tcctgaaaga actgtttgga agtattttgt tcagctttgc
      541 agtgcattgg aacacatgca ttctcgaaga gtcatgcata gagatataaa accagctaat
      601 gtgttcatta cagccactgg ggtggtaaaa cttggagatc ttgggcttgg ccggtttttc
      661 agctcaaaaa ccacagctgc acattcttta gttggtacgc cttattacat gtctccagag
      721 agaatacatg aaaatggata caacttcaaa tctgacatct ggtctcttgg ctgtctacta
      781 tatgagatgg ctgcattaca aagtcctttc tatggtgaca aaatgaattt atactcactg
      841 tgtaagaaga tagaacagtg tgactaccca cctcttcctt cagatcacta ttcagaagaa
      901 ctccgacagt tagttaatat gtgcatcaac ccagatccag agaagcgacc agacgtcacc
      961 tatgtttatg acgtagcaaa gaggatgcat gcatgcactg caagcagctt gccaactttc
     1021 ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc
     1081 actatcagtc aaaataaaat cattatttgc catccagctg atatccccta tagtgagtcg
     1141 tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt
     1201 tacattgcac aagataaaaa tatatcatca tgcctcctct agaccagcca ggacagaaat
     1261 gcctcgactt cgctgctacc caaggttgcc gggtgacgca caccgtggaa acggatgaag
     1321 gcacgaaccc agtggacata agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg
     1381 cgctcacgca actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg
     1441 gttttcatgg cttgttatga ctgttttttt ggggtacagt ctatgcctcg ggcatccaag
     1501 cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg
     1561 cagcagggca gtcgccctaa aacaaagtta aacattatga gggaagcggt gatcgccgaa
     1621 gtatcgactc aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg
     1681 ctggccgtac atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt
     1741 gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac
     1801 gaccttttgg aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc
     1861 accattgttg tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa
     1921 tttggagaat ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac
     1981 attgatctgg ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca
     2041 gcggcggagg aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa
     2101 accttaacgc tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt
     2161 acgttgtccc gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct
     2221 gccgactggg caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctaga
     2281 caggcttatc ttggacaaga agaagatcgc ttggcctcgc gcgcagatca gttggaagaa
     2341 tttgtccact acgtgaaagg cgagatcacc aaggtagtcg gcaaataacc ctcgagccac
     2401 ccatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta
     2461 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2521 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2581 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     2641 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2701 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2761 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2821 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     2881 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     2941 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3001 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3061 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3121 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     3181 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     3241 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     3301 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     3361 acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg
     3421 ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg
     3481 gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt
     3541 tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta
     3601 tttgatgcct ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca
     3661 cgacgttgta aaacgacggc cagtcttaag ctcgggccc
//
