LOCUS       sequence-267450-        4260 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 4260)
  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 4260)
  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-CLK3
FEATURES             Location/Qualifiers
     source          1..4260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     375..394
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     628..645
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(804..831)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(923..1009)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1058..1080
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1072..1089
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1073..1089
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1105..1204
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2675..2774)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2791..2810)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2792..2810)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2815..2831)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2815..2831)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3262..4053
                     /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      join(4146..4260,1..474)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
       61 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
      121 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
      181 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
      241 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
      301 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
      361 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
      421 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
      481 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
      541 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
      601 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
      661 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
      721 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc
      781 caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt
      841 gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa
      901 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
      961 agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag
     1021 ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg ttgtaaaacg
     1081 acggccagtc ttaagctcgg gccccaaata atgattttat tttgactgat agtgacctgt
     1141 tcgttgcaac aaattgatga gcaatgcttt tttataatgc caactttgta caaaaaagtt
     1201 ggcatgcatc actgtaagcg ataccgctcc cctgaaccag acccgtacct gagctaccga
     1261 tggaagagga ggaggtccta cagtcgggaa catgaaggga gactgcgata cccgtcccga
     1321 agggagcctc ccccacgaag atctcggtcc agaagccatg accgcctgcc ctaccagagg
     1381 aggtaccggg agcgccgtga cagcgataca taccggtgtg aagagcggag cccatccttt
     1441 ggagaggact actatggacc ttcacgttct cgtcatcgtc ggcgatcgcg ggagaggggg
     1501 ccataccgga cccgcaagca tgcccaccac tgccacaaac gccgcaccag gtcttgtagc
     1561 agcgcctcct cgagaagcca acagagcagt aagcgcagca gccggagtgt ggaagatgac
     1621 aaggagggtc acctggtgtg ccggatcggc gattggctcc aagagcgata tgagattgtg
     1681 gggaacctgg gtgaaggcac ctttggcaag gtggtggagt gcttggacca tgccagaggg
     1741 aagtctcagg ttgccctgaa gatcatccgc aacgtgggca agtaccggga ggctgcccgg
     1801 ctagaaatca acgtgctcaa aaaaatcaag gagaaggaca aagaaaacaa gttcctgtgt
     1861 gtcttgatgt ctgactggtt caacttccac ggtcacatgt gcatcgcctt tgagctcctg
     1921 ggcaagaaca cctttgagtt cctgaaggag aataacttcc agccttaccc cctaccacat
     1981 gtccggcaca tggcctacca gctctgccac gcccttagat ttctgcatga gaatcagctg
     2041 acccatacag acttgaaacc agagaacatc ctgtttgtga attctgagtt tgaaaccctc
     2101 tacaatgagc acaagagctg tgaggagaag tcagtgaaga acaccagcat ccgagtggct
     2161 gactttggca gtgccacatt tgaccatgag caccacacca ccattgtggc cacccgtcac
     2221 tatcgcccgc ctgaggtgat ccttgagctg ggctgggcac agccctgtga cgtctggagc
     2281 attggctgca ttctctttga gtactaccgg ggcttcacac tcttccagac ccacgaaaac
     2341 cgagagcacc tggtgatgat ggagaagatc ctagggccca tcccatcaca catgatccac
     2401 cgtaccagga agcagaaata tttctacaaa gggggcctag tttgggatga gaacagctct
     2461 gacggccggt atgtgaagga gaactgcaaa cctctgaaga gttacatgct ccaagactcc
     2521 ctggagcacg tgcagctgtt tgacctgatg aggaggatgt tagaatttga ccctgcccag
     2581 cgcatcacac tggccgaggc cctgctgcac cccttctttg ctggcctgac ccctgaggag
     2641 cggtccttcc acaccagccg caacccaagc agatgcccaa ctttcttgta caaagttggc
     2701 attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat
     2761 aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata
     2821 gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
     2881 aaaaatatat catcatgcct cctctagacc agccaggaca gaaatgcctc gacttcgctg
     2941 ctacccaagg ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg aacccagtgg
     3001 acataagcct gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg
     3061 tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt
     3121 tatgactgtt tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac
     3181 gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc
     3241 cctaaaacaa agttaaacat tatgagggaa gcggtgatcg ccgaagtatc gactcaacta
     3301 tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg
     3361 tacggctccg cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg
     3421 gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact
     3481 tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac
     3541 gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag
     3601 cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc
     3661 ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc
     3721 tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg
     3781 aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt
     3841 tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg
     3901 gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga
     3961 caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg
     4021 aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga gccacccatg accaaaatcc
     4081 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4141 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4201 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
//
