LOCUS       sequence-416401-        4380 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 4380)
  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 4380)
  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-GK
FEATURES             Location/Qualifiers
     source          1..4380
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2259..2358)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2375..2394)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2376..2394)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2399..2415)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2399..2415)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2846..3637
                     /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      3730..4318
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4219..4238
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agttggcatg gcagcctcaa agaaggcagt tttggggcca ttggtggggg cggtggacca
      721 gggcaccagt tcgacgcgct ttttggtttt caattcaaaa acagctgaac tacttagtca
      781 tcatcaagta gaaataaaac aagagttccc aagagaagga tgggtggaac aggaccctaa
      841 ggaaattcta cattctgtct atgagtgtat agagaaaaca tgtgagaaac ttggacagct
      901 caaaattgat atttccaaca taaaagctat tggtgtcagc aaccagaggg aaaccactgt
      961 agtctgggac aagataactg gagagcctct ctacaatgct gtggtgtggc ttgatctaag
     1021 aacccagtct accgttgaga gtcttagtaa aagaattaca ggaaataata actttgtcaa
     1081 gtccaagaca ggccttccac ttagcactta cttcagtgca gtgaaacttc gttggctcct
     1141 tgacaatgtg agaaaagttc aaaaggcagt tgaagaaaaa cgagctcttt ttgggactat
     1201 tgattcatgg cttatttgga gtttgacagg aggagtcaat ggaggtgtcc actgtacaga
     1261 tgtaacaaat gcaagtagga ctatgctttt caacattcat tctttggaat gggataaaca
     1321 actctgcgaa ttttttggaa ttccaatgga aattcttcca aatgtccgga gttcttctga
     1381 gatctatggc ctaatgaaaa tctctcatag cgtgaaagct ggggccttgg aaggtgtgcc
     1441 aatatctggg tgtttagggg accagtctgc tgcattggtg ggacaaatgt gcttccagat
     1501 tggacaagcc aaaaatacgt atggaacagg atgtttctta ctatgtaata caggccataa
     1561 gtgtgtattt tctgatcatg gccttctcac cacagtggct tacaaacttg gcagagacaa
     1621 accagtatat tatgctttgg aaggttctgt agctatagct ggtgctgtta ttcgctggct
     1681 aagagacaat cttggaatta taaagacctc agaagaaatt gaaaaacttg ctaaagaagt
     1741 aggtacttct tatggctgct acttcgtccc agcattttcg gggttatatg caccttattg
     1801 ggagcccagc gcaagaggga taatctgtgg actcactcag ttcaccaata aatgccatat
     1861 tgcttttgct gcattagaag ctgtttgttt ccaaactcga gagattttgg atgccatgaa
     1921 tcgagactgt ggaattccac tcagtcattt gcaggtagat ggaggaatga ccagcaacaa
     1981 aattcttatg cagctacaag cagacattct gtatatacca gtagtgaagc cctcaatgcc
     2041 cgaaaccact gcactgggtg cggctatggc ggcaggggct gcagaaggag tcggcgtatg
     2101 gagtctcgaa cccgaggatt tgtctgccgt cacgatggag cggtttgaac ctcagattaa
     2161 tgcggaggaa agtgaaattc gttattctac atggaagaaa gctgtgatga agtcaatggg
     2221 ttgggttaca actcaatctc cagaaagtgg tattccatac ccaactttct tgtacaaagt
     2281 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     2341 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     2401 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     2461 agataaaaat atatcatcat gcctcctcta gaccagccag gacagaaatg cctcgacttc
     2521 gctgctaccc aaggttgccg ggtgacgcac accgtggaaa cggatgaagg cacgaaccca
     2581 gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa
     2641 ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc
     2701 ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg
     2761 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag
     2821 tcgccctaaa acaaagttaa acattatgag ggaagcggtg atcgccgaag tatcgactca
     2881 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
     2941 tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt
     3001 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga
     3061 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
     3121 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
     3181 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc
     3241 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
     3301 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
     3361 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg
     3421 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc
     3481 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
     3541 tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta
     3601 cgtgaaaggc gagatcacca aggtagtcgg caaataaccc tcgagccacc catgaccaaa
     3661 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3721 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3781 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3841 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3901 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3961 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4021 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4081 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4141 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4201 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4261 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4321 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
