LOCUS       sequence-199102-        5808 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 5808)
  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 5808)
  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-PTK2
FEATURES             Location/Qualifiers
     source          1..5808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     22..41
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        22..40
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    58..157
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3176..3275)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3291..3308)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3291..3307)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3300..3322)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      3371..3457
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3549..3576
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3735..3752)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3906..4494)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3986..4005)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4587..5378)
                     /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"
ORIGIN
        1 caggaaacag ctatgaccat gtaatacgac tcactatagg ggatatcagc tggatggcaa
       61 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      121 tttcttataa tgccaacttt gtacaagaaa gttgggtgtg gtctcgtctg cccaagcatt
      181 ttcagtcttg cttggtcaat gacatcgagt aagtttttgg catccacagc cagggcgtga
      241 gcagcagtca gcatttgctt tttgtactct tgctggaggc tggtcatgac atactgctgg
      301 gccagtttca tcttgttgat gagctcaccc aggtcagagt tcaatagctt ctgtgccatc
      361 tcaatctctc ggtgggtgct ggctggtagg aggggaatgg tctcatccac agtggccaat
      421 aatgtcctca gggccaagcc gacttccttc accataggga catactcctc tggtggggct
      481 ggctggattt tactggacat ctcgatgaca gctttcacca ggcccgtcac attctcgtac
      541 accttatcat tcgaccggtc caggttggca gtaggagggg ggctgatttc ctggggctga
      601 agcttgacac cctcgttgta gctgtcagca gggctgctga ggctggcaag gcttcccaga
      661 tgaccgggag ctccagggcg aggcggtttc tttggtggag ctgcaggatc tggtttaccc
      721 acaggctgat atatatgttg gtttccaatc ggaccctgaa gacttccatc ctccctgtca
      781 atactgcctc gagagagtct cacatcaggt ttcagaaatc tttcctcttt ttccagccag
      841 cgctgatctt cttccatttc ctgttgctgt cggattagac gctcttccat cagatgggtt
      901 ggcaacactt gcccaatccc tcgcaggtcc aatactgtag agtcctggta atgattggtt
      961 tgtaccatgt gctgtgggct gggataaaat ccttcgctgg acctcggact gggataaccc
     1021 ggtctgctgg gcttgggcgg tgcttcatca gaccctccgg agtcccagga cactgtggcc
     1081 tgtcttctgg actccatcct catgcgctct tcttgctgag ccttctcttc ctccaggatt
     1141 gtgctgagct gagctttaag ttcagtaaac ctgggccgcc tgctggggtc ataggcccag
     1201 catttcgtca taaggctgta gagggtagga ggacaatttg gaggcattgg taatctttcc
     1261 ccattttcaa ttcgaccgat tacatcattg ttcttcactc cttgaaaagg cttcacacca
     1321 tgcatcagta tctcccacat acacacacca aacatccata cgtcactagc tgaggtaaaa
     1381 cgtcgaaaat tgattgactc tggagccatc catttaatag gcaattttcc tttggaagct
     1441 ttgtagtaag tactatcttc catatatcgg gataatccaa agtctcctaa ttttacacaa
     1501 tcatttgagg acaccagaac attccgagca gcaatgtccc tgtgtacaaa tcttttgctc
     1561 tctagatatg caagagctgt actaagctga taggcataca ggatcaaaga tgctagatcc
     1621 aaactgtatt tccttacttg caaaaatgac ctcagctctc caagtgtgca cagctccatg
     1681 attatccaga caggattctc tgtgatgact ccaatcagct tcacaatatg aggatggtca
     1741 aactgacgca ttgttaaggc ttcttgaaga aatttctctc tcacgctgtc cgaagtacag
     1801 tttttacatg ttttaattgc aaccgccaaa gctggattct ctggactcat ataaatgcct
     1861 tgatgtacat ctccaaattg gccttctcca atacatcgtc caagttctat tctttctctt
     1921 tgaatctcat aatccctggt tgagggcatg gtgtaagtat cttcttcatc tataatctca
     1981 gcataatcat ctgtttctga cacagagacg gcgtgtgtcc gcatgccttg cttttcgctg
     2041 ttggccaact ttggtattga tggcaaagcc cgttcacctt ctttctgagg tctgatgata
     2101 aatgactgcg aggttccatt caccagccgg cagtacccat ctattaggtc agccatattc
     2161 tccgcaatgg ttagggatgg tgccgtcact gtcagaggct cgggtgcacc tgctattttt
     2221 agttgtagca ttccttttct gtccttgtct tcactgtttg aatactgaat ggtttgcact
     2281 tgagtgaagt cagcaagatg tgtgggattg cagcccttgt ccgttaggta actgattcct
     2341 tcttctgggc cgattgccag ttccactgaa ataatccagc ttgaaccaag agcacacttg
     2401 aagcattcct tatcaaatct gtagactgga gacaggatct caaagaattt cagaatactt
     2461 tcttctctat taaggttggc aaattgtcta aatgtttgtt ggatcagttt tcttagtgtt
     2521 ttggccttga cagaatccag taaactctta ggaaaaaatc gctttaaacc aacatctttt
     2581 tctaatactt catagttaga cttcttttct agtgcattgc cccgcatctc ccagtatgat
     2641 cgccgtattt ctagacaacc caacttcaaa gcaatttcct ggtccacttg atcagctatc
     2701 tctaacatat aatcgctctt cacctgttga tagaagaaat tcaaagttgg cttatcttca
     2761 gtaaactggt ttagaaatcc ttttggcaaa taacgaattc tcaattcata tttccactcc
     2821 tctggtgggt gagcaagctc atacttctcc ctcacactgg agacgcccat atccacgtga
     2881 agccagtgaa cctcctctga ccgcaggtga ctgaggcgga atccatagca ggccacatgc
     2941 tttactttgt gactgtccac tatcttctga atgatgcccc tgacatcagt agcatctcca
     3001 tgcctgataa tactggccca ggtggttggc tcactattgc tttcaaaata atgaaagacc
     3061 tttaatactc gctccattgc accaggagaa cgttccatac cagtacccag gtgagtctta
     3121 gtactcgaat ttggtgtgtg attcaagttg gggtcaaggt aagcagctgc catggtgcca
     3181 acttttttgt acaaagttgg cattataaaa aagcattgct catcaatttg ttgcaacgaa
     3241 caggtcacta tcagtcaaaa taaaatcatt atttggggcc cgagcttaag actggccgtc
     3301 gttttacaac gtcgtgactg ggaaaacatc catgctagcg ttaacgcgag agtagggaac
     3361 tgccaggcat caaataaaac gaaaggctca gtcggaagac tgggcctttc gttttatctg
     3421 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt
     3481 tgtgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca
     3541 aactaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcttcctgg
     3601 ctagcggtac gcgtattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3661 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3721 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3781 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3841 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3901 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3961 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4021 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4081 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4141 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4201 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4261 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4321 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4381 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4441 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4501 cctttgatct tttctacggg gtctgacgct cagtggaacg acgcgtaact cacgttaagg
     4561 gattttggtc atgggtggct cgagggttat ttgccgacta ccttggtgat ctcgcctttc
     4621 acgtagtgga caaattcttc caactgatct gcgcgcgagg ccaagcgatc ttcttcttgt
     4681 ccaagataag cctgtctagc ttcaagtatg acgggctgat actgggccgg caggcgctcc
     4741 attgcccagt cggcagcgac atccttcggc gcgattttgc cggttactgc gctgtaccaa
     4801 atgcgggaca acgtaagcac tacatttcgc tcatcgccag cccagtcggg cggcgagttc
     4861 catagcgtta aggtttcatt tagcgcctca aatagatcct gttcaggaac cggatcaaag
     4921 agttcctccg ccgctggacc taccaaggca acgctatgtt ctcttgcttt tgtcagcaag
     4981 atagccagat caatgtcgat cgtggctggc tcgaagatac ctgcaagaat gtcattgcgc
     5041 tgccattctc caaattgcag ttcgcgctta gctggataac gccacggaat gatgtcgtcg
     5101 tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc tctctccagg ggaagccgaa
     5161 gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt catcaagcct tacggtcacc
     5221 gtaaccagca aatcaatatc actgtgtggc ttcaggccgc catccactgc ggagccgtac
     5281 aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga tgacgccaac tacctctgat
     5341 agttgagtcg atacttcggc gatcaccgct tccctcataa tgtttaactt tgttttaggg
     5401 cgactgccct gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg
     5461 taacgcgctt gctgcttgga tgcccgaggc atagactgta ccccaaaaaa acagtcataa
     5521 caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac
     5581 cagttgcgtg agcgcatacg ctacttgcat tacagcttac gaaccgaaca ggcttatgtc
     5641 cactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggtagc
     5701 agcgaagtcg aggcatttct gtcctggctg gtctagagga ggcatgatga tatattttta
     5761 tcttgtgcaa tgtaacatca gagattttga gacacgggcc agagctgc
//
