LOCUS       sequence-217148-        3972 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 3972)
  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 3972)
  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-MAP2K1
FEATURES             Location/Qualifiers
     source          1..3972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(157..184)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(276..362)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     411..433
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     425..442
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     426..442
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    458..557
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1740..1839)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1856..1875)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1857..1875)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1880..1896)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1880..1896)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2327..3118
                     /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      3211..3799
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3700..3719
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3953..3970
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
       61 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac
      121 gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc
      181 ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc
      241 cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc
      301 accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt
      361 tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg
      421 acgttgtaaa acgacggcca gtcttaagct cgggccccaa ataatgattt tattttgact
      481 gatagtgacc tgttcgttgc aacaaattga tgagcaatgc ttttttataa tgccaacttt
      541 gtacaaaaaa gttggcacca tgcccaagaa gaagccgacg cccatccagc tgaacccggc
      601 ccccgacggc tctgcagtta acgggaccag ctctgcggag accaacttgg aggccttgca
      661 gaagaagctg gaggagctag agcttgatga gcagcagcga aagcgccttg aggcctttct
      721 tacccagaag cagaaggtgg gagaactgaa ggatgacgac tttgagaaga tcagtgagct
      781 gggggctggc aatggcggtg tggtgttcaa ggtctcccac aagccttctg gcctggtcat
      841 ggccagaaag ctaattcatc tggagatcaa acccgcaatc cggaaccaga tcataaggga
      901 gctgcaggtt ctgcatgagt gcaactctcc gtacatcgtg ggcttctatg gtgcgttcta
      961 cagcgatggc gagatcagta tctgcatgga gcacatggat ggaggttctc tggatcaagt
     1021 cctgaagaaa gctggaagaa ttcctgaaca aattttagga aaagttagca ttgctgtaat
     1081 aaaaggcctg acatatctga gggagaagca caagatcatg cacagagatg tcaagccctc
     1141 caacatccta gtcaactccc gtggggagat caagctctgt gactttgggg tcagcgggca
     1201 gctcatcgac tccatggcca actccttcgt gggcacaagg tcctacatgt cgccagaaag
     1261 actccagggg actcattact ctgtgcagtc agacatctgg agcatgggac tgtctctggt
     1321 agagatggcg gttgggaggt atcccatccc tcctccagat gccaaggagc tggagctgat
     1381 gtttgggtgc caggtggaag gagatgcggc tgagacccca cccaggccaa ggacccccgg
     1441 gaggcccctt agctcatacg gaatggacag ccgacctccc atggcaattt ttgagttgtt
     1501 ggattacata gtcaacgagc ctcctccaaa actgcccagt ggagtgttca gtctggaatt
     1561 tcaagatttt gtgaataaat gcttaataaa aaaccccgca gagagagcag atttgaagca
     1621 actcatggtt catgctttta tcaagagatc tgatgctgag gaagtggatt ttgcaggttg
     1681 gctctgctcc accatcggcc ttaaccagcc cagcacacca acccatgctg ctggcgtctt
     1741 gccaactttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
     1801 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg atatccccta
     1861 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
     1921 tctctgatgt tacattgcac aagataaaaa tatatcatca tgcctcctct agaccagcca
     1981 ggacagaaat gcctcgactt cgctgctacc caaggttgcc gggtgacgca caccgtggaa
     2041 acggatgaag gcacgaaccc agtggacata agcctgttcg gttcgtaagc tgtaatgcaa
     2101 gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg gtggtaacgg
     2161 cgcagtggcg gttttcatgg cttgttatga ctgttttttt ggggtacagt ctatgcctcg
     2221 ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa
     2281 cgatgttacg cagcagggca gtcgccctaa aacaaagtta aacattatga gggaagcggt
     2341 gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc gccatctcga
     2401 accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc tgaagccaca
     2461 cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc
     2521 tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga ttctccgcgc
     2581 tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc cagctaagcg
     2641 cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct tcgagccagc
     2701 cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata gcgttgcctt
     2761 ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc tatttgaggc
     2821 gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg atgagcgaaa
     2881 tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa tcgcgccgaa
     2941 ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc ccgtcatact
     3001 tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc gcgcagatca
     3061 gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg gcaaataacc
     3121 ctcgagccac ccatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     3181 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3241 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3301 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3361 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3421 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3481 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3541 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3601 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3661 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3721 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3781 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3841 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3901 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3961 agtgagcgag ga
//
