LOCUS       sequence-401866-        4563 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 4563)
  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 4563)
  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-PAK4
FEATURES             Location/Qualifiers
     source          1..4563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..37
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     29..46
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     30..46
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    62..161
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1935..2034)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2051..2070)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2052..2070)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2075..2091)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2075..2091)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2522..3313
                     /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      3406..3994
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3895..3914
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4148..4165
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4324..4351)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4443..4529)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 agcatggatg ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc
       61 ccaaataatg attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgatgagca
      121 atgctttttt ataatgccaa ctttgtacaa aaaagttggc atgtttggga agaggaagaa
      181 gcgggtggag atctccgcgc cgtccaactt cgagcaccgc gtgcacacgg gcttcgacca
      241 gcacgagcag aagttcacgg ggctgccccg ccagtggcag agcctgatcg aggagtcggc
      301 tcgccggccc aagcccctcg tcgaccccgc ctgcatcacc tccatccagc ccggggcccc
      361 caagaccatc gtgcggggca gcaaaggtgc caaagatggg gccctcacgc tgctgctgga
      421 cgagtttgag aacatgtcgg tgacacgctc caactccctg cggagagaca gcccgccgcc
      481 gcccgcccgt gcccgccagg aaaatgggat gccagaggag ccggccacca cggccagagg
      541 gggcccaggg aaggcaggca gccgaggccg gttcgccggt cacagcgagg cgggtggcgg
      601 cagtggtgac aggcgacggg cggggccaga gaagaggccc aagtcttcca gggagggctc
      661 agggggtccc caggagtcct cccgggacaa acgccccctc tccgggcctg atgtcggcac
      721 cccccagcct gctggtctgg ccagtggggc gaaactggca gctggccggc cctttaacac
      781 ctacccgagg gctgacacgg accacccatc ccggggtgcc cagggggagc ctcatgacgt
      841 ggcccctaac gggccatcag cggggggcct ggccatcccc cagtcctcct cctcctcctc
      901 ccggcctccc acccgagccc gaggtgcccc cagccctgga gtgctgggac cccacgcctc
      961 agagccccag ctggcccctc cagcctgcac ccccgccgcc cctgctgttc ctgggccccc
     1021 tggcccccgc tcaccacagc gggagccaca gcgagtatcc catgagcagt tccgggctgc
     1081 cctgcagctg gtggtggacc caggcgaccc ccgctcctac ctggacaact tcatcaagat
     1141 tggcgagggc tccacgggca tcgtgtgcat cgccaccgtg cgcagctcgg gcaagctggt
     1201 ggccgtcaag aagatggacc tgcgcaagca gcagaggcgc gagctgctct tcaacgaggt
     1261 ggtaatcatg agggactacc agcacgagaa tgtggtggag atgtacaaca gctacctggt
     1321 gggggacgag ctctgggtgg tcatggagtt cctggaagga ggcgccctca ccgacatcgt
     1381 cacccacacc aggatgaacg aggagcagat cgcggccgtg tgccttgcag tgctgcaggc
     1441 cctgtcggtg ctccacgccc agggcgtcat ccaccgggac atcaagagcg actcgatcct
     1501 gctgacccat gatggcaggg tgaagctgtc agactttggg ttctgcgccc aggtgagcaa
     1561 ggaagtgccc cgaaggaagt cgctggtcgg cacgccctac tggatggccc cagagctcat
     1621 ctcccgcctt ccctacgggc cagaggtaga catctggtcg ctggggataa tggtgattga
     1681 gatggtggac ggagagcccc cctacttcaa cgagccaccc ctcaaagcca tgaagatgat
     1741 tcgggacaac ctgccacccc gactgaagaa cctgcacaag gtgtcgccat ccctgaaggg
     1801 cttcctggac cgcctgctgg tgcgagaccc tgcccagcgg gccacggcag ccgagctgct
     1861 gaagcaccca ttcctggcca aggcagggcc gcctgccagc atcgtgcccc tcatgcgcca
     1921 gaaccgcacc agatgcccaa ctttcttgta caaagttggc attataagaa agcattgctt
     1981 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     2041 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     2101 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgcct
     2161 cctctagacc agccaggaca gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg
     2221 acgcacaccg tggaaacgga tgaaggcacg aacccagtgg acataagcct gttcggttcg
     2281 taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg
     2341 cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt
     2401 acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat
     2461 gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat
     2521 tatgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat
     2581 cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg
     2641 cggcctgaag ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga
     2701 aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag
     2761 cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg
     2821 ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg
     2881 tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga
     2941 acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca
     3001 ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc
     3061 tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg
     3121 caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta
     3181 tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc
     3241 ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt
     3301 agtcggcaaa taaccctcga gccacccatg accaaaatcc cttaacgtga gttacgcgtc
     3361 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3421 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3481 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3541 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3601 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3661 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3721 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3781 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3841 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3901 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3961 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4021 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     4081 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     4141 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     4201 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     4261 gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc caggaagagt ttgtagaaac
     4321 gcaaaaaggc catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc
     4381 gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc
     4441 ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag
     4501 tcttccgact gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcgttaac
     4561 gct
//
