LOCUS       sequence-217975-        4068 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 4068)
  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 4068)
  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-MAP2K7
FEATURES             Location/Qualifiers
     source          1..4068
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(143..170)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(262..348)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     397..419
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     411..428
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     412..428
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    444..543
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1822..1921)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1938..1957)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1939..1957)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1962..1978)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1962..1978)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2409..3200
                     /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      3293..3881
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3782..3801
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4035..4052
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
       61 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc
      121 aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg
      181 atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac
      241 gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
      301 gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt
      361 tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt tgtaaaacga
      421 cggccagtct taagctcggg ccccaaataa tgattttatt ttgactgata gtgacctgtt
      481 cgttgcaaca aattgatgag caatgctttt ttataatgcc aactttgtac aaaaaagttg
      541 gcatggcggc gtcctccctg gaacagaagc tgtcccgcct ggaagcaaag ctaaagcagg
      601 agaaccggga ggcccggcgg aggatcgacc tcaacctgga tatcagcccc cagcggccca
      661 ggcccaccct gcagctcccg ctggccaacg atgggggcag ccgctcgcca tcctcagaga
      721 gctccccgca gcaccccacg ccccccgccc ggccccgcca catgctgggg ctcccgtcaa
      781 ccctgttcac accccgcagc atggagagca ttgagattga ccagaagctg caggagatca
      841 tgaagcagac gggctacctg accatcgggg gccagcgcta ccaggcagaa atcaacgacc
      901 tggagaactt gggcgagatg ggcagcggca cctgcggcca ggtgtggaag atgcgcttcc
      961 ggaagaccgg ccacgtcatt gccgttaagc aaatgcggcg ctccgggaac aaggaggaga
     1021 acaagcgcat cctcatggac ctggatgtgg tgctgaagag ccacgactgc ccctacatcg
     1081 tgcagtgctt tgggacgttc atcaccaaca cggatgtctt catcgccatg gagctcatgg
     1141 gcacctgcgc tgagaagctc aagaagcgga tgcagggccc catccccgag cgcattctgg
     1201 gcaagatgac agtggcgatt gtgaaggcgc tgtactacct gaaggagaag cacggtgtca
     1261 tccaccgcga cgtcaagccc tccaacatcc tgctggacga gcggggccag atcaagctct
     1321 gcgacttcgg catcagcggc cgcctggtgg actccaaagc caagacgcgg agcgccggct
     1381 gtgccgccta catggcaccc gagcgcattg accccccaga ccccaccaag ccggactatg
     1441 acatccgggc cgacgtatgg agcctgggca tctcgttgcc ctgcccgtct ccctcccagg
     1501 tggagctggc aacaggacag tttccctaca agaactgcaa gacggacttt gaggtcctca
     1561 ccaaagtcct acaggaagag cccccgcttc tgcccggaca catgggcttc tcgggggact
     1621 tccagtcctt cgtcaaagac tgccttacta aagatcacag gaagagacca aagtataata
     1681 agctacttga acacagcttc atcaagcgct acgagacgct ggaggtggac gtggcgtcct
     1741 ggttcaagga tgtcatggcg aagactgagt caccgcggac tagcggcgtc ctgagccagc
     1801 cccacctgcc cttcctcagg tacccaactt tcttgtacaa agttggcatt ataagaaagc
     1861 attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt
     1921 gccatccagc tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc
     1981 agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
     2041 catgcctcct ctagaccagc caggacagaa atgcctcgac ttcgctgcta cccaaggttg
     2101 ccgggtgacg cacaccgtgg aaacggatga aggcacgaac ccagtggaca taagcctgtt
     2161 cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga
     2221 ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt
     2281 ttggggtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat
     2341 gtttgatgtt atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt
     2401 taaacattat gagggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg
     2461 gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag
     2521 tggatggcgg cctgaagcca cacagtgata ttgatttgct ggttacggtg accgtaaggc
     2581 ttgatgaaac aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg
     2641 gagagagcga gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc
     2701 cgtggcgtta tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc
     2761 ttgcaggtat cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag
     2821 caagagaaca tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc
     2881 ctgaacagga tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg
     2941 actgggctgg cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag
     3001 taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg
     3061 cccagtatca gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc
     3121 gcttggcctc gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa ggcgagatca
     3181 ccaaggtagt cggcaaataa ccctcgagcc acccatgacc aaaatccctt aacgtgagtt
     3241 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3301 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3361 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3421 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3481 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3541 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3601 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3661 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3721 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3781 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3841 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3901 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     3961 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4021 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcc
//
