LOCUS       sequence-170545-        4968 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 4968)
  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 4968)
  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-ITK
FEATURES             Location/Qualifiers
     source          1..4968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(116..143)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(235..321)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     370..392
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     384..401
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     385..401
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    417..516
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2377..2476)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2493..2512)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2494..2512)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2517..2533)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2517..2533)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3282..4073
                     /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      4166..4754
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4655..4674
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4908..4925
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
       61 agcgcaacgc aattaatacg cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag
      121 gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc
      181 tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt
      241 cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtcttccga
      301 ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcgtta acgctagcat
      361 ggatgttttc ccagtcacga cgttgtaaaa cgacggccag tcttaagctc gggccccaaa
      421 taatgatttt attttgactg atagtgacct gttcgttgca acaaattgat gagcaatgct
      481 tttttataat gccaactttg tacaaaaaag ttggcaccat gaacaacttt atcctcctgg
      541 aagaacagct catcaagaaa tcccaacaaa agagaagaac ttctccctcg aactttaaag
      601 tccgcttctt tgtgttaacc aaagccagcc tggcatactt tgaagatcgt catgggaaga
      661 agcgcacgct gaaggggtcc attgagctct cccgaatcaa atgtgttgag attgtgaaaa
      721 gtgacatcag catcccatgc cactataaat acccgtttca ggtggtgcat gacaactacc
      781 tcctatatgt gtttgctcca gatcgtgaga gccggcagcg ctgggtgctg gcccttaaag
      841 aagaaacgag gaataataac agtttggtgc ctaaatatca tcctaatttc tggatggatg
      901 ggaagtggag gtgctgttct cagctggaga agcttgcaac aggctgtgcc caatatgatc
      961 caaccaagaa tgcttcaaag aagcctcttc ctcctactcc tgaagacaac aggcgaccac
     1021 tttgggaacc tgaagaaact gtggtcattg ccttatatga ctaccaaacc aatgatcctc
     1081 aggaactcgc actgcggcgc aacgaagagt actgcctgct ggacagttct gagattcact
     1141 ggtggagagt ccaggacagg aatgggcatg aaggatatgt accaagcagt tatctggtgg
     1201 aaaaatctcc aaataatctg gaaacctatg agtggtacaa taagagtatc agccgagaca
     1261 aagctgaaaa acttcttttg gacacaggca aagaaggagc cttcatggta agggattcca
     1321 ggactgcagg aacatacacc gtgtctgttt tcaccaaggc tgttgtaagt gagaacaatc
     1381 cctgtataaa gcattatcac atcaaggaaa caaatgacaa tcctaagcga tactatgtgg
     1441 ctgaaaagta tgtgttcgat tccatccctc ttctcatcaa ctatcaccaa cataatggag
     1501 gaggcctggt gactcgactc cggtatccag tttgttttgg gaggcagaaa gccccagtta
     1561 cagcagggct gagatacggg aaatgggtga tcgacccctc agagctcact tttgtccaag
     1621 agattggcag tgggcaattt gggttggtgc atctgggcta ctggctcaac aaggacaagg
     1681 tggctatcaa aaccattcgg gaaggggcta tgtcagaaga ggacttcata gaggaggctg
     1741 aagtaatgat gaaactctct catcccaaac tggtgcagct gtatggggtg tgcctggagc
     1801 aggcccccat ctgcctggtg tttgagttca tggagcacgg ctgcctgtca gattatctac
     1861 gcacccagcg gggacttttt gctgcagaga ccctgctggg catgtgtctg gatgtgtgtg
     1921 agggcatggc ctacctggaa gaggcatgtg tcatccacag agacttggct gccagaaatt
     1981 gtttggtggg agaaaaccaa gtcatcaagg tgtctgactt tgggatgaca aggttcgttc
     2041 tggatgatca gtacaccagt tccacaggca ccaaattccc ggtgaagtgg gcatccccag
     2101 aggttttctc tttcagtcgc tatagcagca agtccgatgt gtggtcattt ggtgtgctga
     2161 tgtgggaagt tttcagtgaa ggcaaaatcc cgtatgaaaa ccgaagcaac tcagaggtgg
     2221 tggaagacat cagtaccgga tttcggttgt acaagccccg gctggcctcc acacacgtct
     2281 accagattat gaatcactgc tggaaagaga gaccagaaga tcggccagcc ttctccagac
     2341 tgctgcgtca actggctgaa attgcagaat caggacttcc aactttcttg tacaaagttg
     2401 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
     2461 ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca
     2521 tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
     2581 ataaaaatat atcatcatgc ctcctctaga ccagccagga cagaaatgcc tcgacttcgc
     2641 tgctacccaa ggttgccggg tgacgcacac cgtggaaacg gatgaaggca cgaacccagt
     2701 ggacataagc ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact
     2761 ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt
     2821 gttatgactg tttttttggg gtacagtcta tgccgctctg gcccgtgtct caaaatctct
     2881 gatgttacat tgcacaagat aaaaatatat catcatgcct cctctagacc agccaggaca
     2941 gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg acgcacaccg tggaaacgga
     3001 tgaaggcacg aacccagtgg acataagcct gttcggttcg taagctgtaa tgcaagtagc
     3061 gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag
     3121 tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat
     3181 ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg
     3241 ttacgcagca gggcagtcgc cctaaaacaa agttaaacat tatgagggaa gcggtgatcg
     3301 ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga
     3361 cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg
     3421 atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga
     3481 tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag
     3541 aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac
     3601 tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga
     3661 tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag
     3721 gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa
     3781 atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag
     3841 tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg
     3901 tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag
     3961 ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg
     4021 aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga
     4081 gccacccatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc
     4141 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4201 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4261 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4321 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4381 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4441 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4501 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4561 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4621 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4681 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4741 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4801 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4861 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4921 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgc
//
