LOCUS       sequence-329334-        3756 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 3756)
  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 3756)
  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-PBK
FEATURES             Location/Qualifiers
     source          1..3756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(562..589)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(681..767)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     816..838
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     830..847
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     831..847
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    863..962
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1929..2028)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2045..2064)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2046..2064)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2069..2085)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2069..2085)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2516..3307
                     /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      join(3400..3756,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca
      541 ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga
      601 tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg
      661 ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag
      721 ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt
      781 ccctactctc gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac
      841 ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag tgacctgttc
      901 gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca aaaaagttgg
      961 catggaaggg atcagtaatt tcaagacacc aagcaaatta tcagaaaaaa agaaatctgt
     1021 attatgttca actccaacta taaatatccc ggcctctcca tttatgcaga agcttggctt
     1081 tggtactggg gtaaatgtgt acctaatgaa aagatctcca agaggtttgt ctcattctcc
     1141 ttgggctgta aaaaagatta atcctatatg taatgatcat tatcgaagtg tgtatcaaaa
     1201 gagactaatg gatgaagcta agattttgaa aagccttcat catccaaaca ttgttggtta
     1261 tcgtgctttt actgaagcca atgatggcag tctgtgtctt gctatggaat atggaggtga
     1321 aaagtctcta aatgacttaa tagaagaacg atataaagcc agccaagatc cttttccagc
     1381 agccataatt ttaaaagttg ctttgaatat ggcaagaggg ttaaagtatc tgcaccaaga
     1441 aaagaaactg cttcatggag acataaagtc ttcaaatgtt gtaattaaag gcgattttga
     1501 aacaattaaa atctgtgatg taggagtctc tctaccactg gatgaaaata tgactgtgac
     1561 tgaccctgag gcttgttaca ttggcacaga gccatggaaa cccaaagaag ctgtggagga
     1621 gaatggtgtt attactgaca aggcagacat atttgccttt ggccttactt tgtgggaaat
     1681 gatgacttta tcgattccac acattaatct ttcaaatgat gatgatgatg aagataaaac
     1741 ttttgatgaa agtgattttg atgatgaagc atactatgca gcgttgggaa ctaggccacc
     1801 tattaatatg gaagaactgg atgaatcata ccagaaagta attgaactct tctctgtatg
     1861 cactaatgaa gaccctaaag atcgtccttc tgctgcacac attgttgaag ctctggaaac
     1921 agatgtctac ccaactttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     1981 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     2041 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     2101 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gcctcctcta
     2161 gaccagccag gacagaaatg cctcgacttc gctgctaccc aaggttgccg ggtgacgcac
     2221 accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct
     2281 gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg
     2341 tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc
     2401 tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg
     2461 gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa acattatgag
     2521 ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg
     2581 ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct
     2641 gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac
     2701 gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat
     2761 tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc
     2821 agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt
     2881 cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag
     2941 cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct
     3001 atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga
     3061 tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat
     3121 cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc
     3181 cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg
     3241 cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg
     3301 caaataaccc tcgagccacc catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     3361 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3421 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3481 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3541 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3601 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3661 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3721 ggggggttcg tgcacacagc ccagcttgga gcgaac
//
