LOCUS       sequence-457346-        4062 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 4062)
  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 4062)
  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-CSNK1G3
FEATURES             Location/Qualifiers
     source          1..4062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1373..1472)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1489..1508)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1490..1508)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1513..1529)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1513..1529)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1960..2751
                     /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      2844..3432
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3333..3352
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3586..3603
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3762..3789)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3881..3967)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4016..4038
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4030..4047
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4031..4047
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagttggca ccatggaaaa taaaaagaaa
      121 gacaaggaca aatcagatga tagaatggca cgacctagtg gtcgatcggg acacaacact
      181 cgaggaactg ggtcttcatc gtctggagtt ttaatggttg gacctaactt tagagttgga
      241 aaaaaaattg gatgtggcaa ttttggagaa ttacgattag ggaaaaattt atacacaaat
      301 gaatatgtgg caattaagtt ggagcccatg aaatcaagag caccacagct acatttggaa
      361 tacagattct ataagcagtt aggatctgga gatggtatac ctcaagttta ctatttcggc
      421 ccttgtggta aatacaatgc tatggtgctg gaactgctgg gacctagttt ggaagacttg
      481 tttgacttgt gtgacagaac attttctctt aaaacagttc tcatgatagc tatacaactg
      541 atttctcgca tggaatatgt ccattcaaag aacttgatat acagagatgt aaaacctgag
      601 aacttcttaa taggacgacc aggaaacaaa acccagcaag ttattcacat tatagatttt
      661 ggtttggcaa aggaatatat tgatccggag acaaagaaac acataccata cagagaacac
      721 aagagcctta caggaacagc tagatatatg agcataaaca cacatttagg aaaagaacaa
      781 agtagaagag acgatttaga agctttaggt catatgttca tgtattttct gagaggcagt
      841 cttccttggc aaggcttaaa ggctgacaca ttaaaggaga ggtatcagaa aattggagat
      901 acaaaacggg ctacaccaat agaagtgtta tgtgaaaatt ttccagaaat ggcaacatat
      961 cttcgttatg taagaaggct agattttttt gaaaaaccag actatgacta cttaagaaag
     1021 ctttttactg acttgtttga tcgaaaagga tatatgtttg attatgaata tgactggatt
     1081 ggtaaacagt tgcctactcc agtgggtgca gttcagcaag atcctgctct gtcatcaaac
     1141 agagaagcac atcaacacag agataagatg caacaatcca aaaaccaggt tgtaagttct
     1201 acaaatggag agttaaacac agatgacccc accgcaggac gttcaaatgc acccatcaca
     1261 gcccctactg aagtagaagt gatggatgaa accaactgcc agaaagtgtt gaacatgtgg
     1321 tgctgctgtt ttttcaaacg aaggaaaagg aaaaccatac agcgccacaa attgccaact
     1381 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     1441 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag
     1501 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     1561 tgttacattg cacaagataa aaatatatca tcatgcctcc tctagaccag ccaggacaga
     1621 aatgcctcga cttcgctgct acccaaggtt gccgggtgac gcacaccgtg gaaacggatg
     1681 aaggcacgaa cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt
     1741 atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg
     1801 gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc
     1861 aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt
     1921 acgcagcagg gcagtcgccc taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc
     1981 gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg
     2041 ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat
     2101 attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc
     2161 aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa
     2221 gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg
     2281 caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc
     2341 gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt
     2401 ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat
     2461 gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg
     2521 cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc
     2581 gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct
     2641 agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa
     2701 gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata accctcgagc
     2761 cacccatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     2821 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2881 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2941 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3001 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3061 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3121 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3181 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3241 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3301 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3361 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3421 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3481 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3541 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3601 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     3661 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     3721 aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga
     3781 tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
     3841 cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     3901 cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt
     3961 ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag
     4021 tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cc
//
