LOCUS       sequence-267448-        3729 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 3729)
  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 3729)
  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-PIM1
FEATURES             Location/Qualifiers
     source          1..3729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(940..1039)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1056..1075)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1057..1075)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1080..1096)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1080..1096)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1527..2318
                     /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      2411..2999
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2900..2919
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3153..3170
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3329..3356)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3448..3534)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3583..3605
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3597..3614
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3598..3614
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3630..3729
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 tgctcttgtc caaaatcaac tcgcttgccc acctgcgcgc cgcgccctgc aacgacctgc
       61 acgccaccaa gctggcgccc ggcaaggaga aggagcccct ggagtcgcag taccaggtgg
      121 gcccgctact gggcagcggc ggcttcggct cggtctactc aggcatccgc gtctccgaca
      181 acttgccggt ggccatcaaa cacgtggaga aggaccggat ttccgactgg ggagagctgc
      241 ctaatggcac tcgagtgccc atggaagtgg tcctgctgaa gaaggtgagc tcgggtttct
      301 ccggcgtcat taggctcctg gactggttcg agaggcccga cagtttcgtc ctgatcctgg
      361 agaggcccga gccggtgcaa gatctcttcg acttcatcac ggaaagggga gccctgcaag
      421 aggagctggc ccgcagcttc ttctggcagg tgctggaggc cgtgcggcac tgccacaact
      481 gcggggtgct ccaccgcgac atcaaggacg aaaacatcct tatcgacctc aatcgcggcg
      541 agctcaagct catcgacttc gggtcggggg cgctgctcaa ggacaccgtc tacacggact
      601 tcgatgggac ccgagtgtat agccctccag agtggatccg ctaccatcgc taccatggca
      661 ggtcggcggc agtctggtcc ctggggatcc tgctgtatga tatggtgtgt ggagatattc
      721 ctttcgagca tgacgaagag atcatcaggg gccaggtttt cttcaggcag agggtctctt
      781 cagaatgtca gcatctcatt agatggtgct tggccctgag accatcagat aggccaacct
      841 tcgaagaaat ccagaaccat ccatggatgc aagatgttct cctgccccag gaaactgctg
      901 agatccacct ccacagcctg tcgccggggc ccagcaaata cccaactttc ttgtacaaag
      961 ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
     1021 aaaataaaat cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg
     1081 tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac
     1141 aagataaaaa tatatcatca tgcctcctct agaccagcca ggacagaaat gcctcgactt
     1201 cgctgctacc caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc
     1261 agtggacata agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca
     1321 actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg
     1381 cttgttatga ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc
     1441 gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca
     1501 gtcgccctaa aacaaagtta aacattatga gggaagcggt gatcgccgaa gtatcgactc
     1561 aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac
     1621 atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg
     1681 ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg
     1741 aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg
     1801 tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat
     1861 ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg
     1921 ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg
     1981 aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc
     2041 tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc
     2101 gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg
     2161 caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc
     2221 ttggacaaga agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact
     2281 acgtgaaagg cgagatcacc aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa
     2341 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
     2401 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2461 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2521 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     2581 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2641 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     2701 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     2761 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     2821 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     2881 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2941 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3001 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3061 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3121 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3181 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     3241 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
     3301 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
     3361 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
     3421 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
     3481 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
     3541 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
     3601 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
     3661 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
     3721 aagttggca
//
