LOCUS       sequence-345437-        3822 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 3822)
  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 3822)
  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-AURKB
FEATURES             Location/Qualifiers
     source          1..3822
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             864..875
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(1165..1264)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1281..1300)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1282..1300)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1305..1321)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1305..1321)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1752..2543
                     /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      2636..3224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3125..3144
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3378..3395
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3554..3581)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3673..3759)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3808..3822,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3822,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagttgg catggcccag aaggagaact cctacccctg gccctacggc cgacagacgg
      181 ctccatctgg cctgagcacc ctgccccagc gagtcctccg gaaagagcct gtcaccccat
      241 ctgcacttgt cctcatgagc cgctccaatg tccagcccac agctgcccct ggccagaagg
      301 tgatggagaa tagcagtggg acacccgaca tcttaacgcg gcacttcaca attgatgact
      361 ttgagattgg gcgtcctctg ggcaaaggca agtttggaaa cgtgtacttg gctcgggaga
      421 agaaaagcca tttcatcgtg gcgctcaagg tcctcttcaa gtcccagata gagaaggagg
      481 gcgtggagca tcagctgcgc agagagatcg aaatccaggc ccacctgcac catcccaaca
      541 tcctgcgtct ctacaactat ttttatgacc ggaggaggat ctacttgatt ctagagtatg
      601 ccccccgcgg ggagctctac aaggagctgc agaagagctg cacatttgac gagcagcgaa
      661 cagccacgat catggaggag ttggcagatg ctctaatgta ctgccatggg aagaaggtga
      721 ttcacagaga cataaagcca gaaaatctgc tcttagggct caagggagag ctgaagattg
      781 ctgacttcgg ctggtctgtg catgcgccct ccctgaggag gaagacaatg tgtggcaccc
      841 tggactacct gcccccagag atgattgagg ggcgcatgca caatgagaag gtggatctgt
      901 ggtgcattgg agtgctttgc tatgagctgc tggtggggaa cccacccttt gagagtgcat
      961 cacacaacga gacctatcgc cgcatcgtca aggtggacct aaagttcccc gcttccgtgc
     1021 ccatgggagc ccaggacctc atctccaaac tgctcaggca taacccctcg gaacggctgc
     1081 ccctggccca ggtctcagcc cacccttggg tccgggccaa ctctcggagg gtgctgcctc
     1141 cctctgccct tcaatctgtc gcctgcccaa ctttcttgta caaagttggc attataagaa
     1201 agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta
     1261 tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct
     1321 ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     1381 catcatgcct cctctagacc agccaggaca gaaatgcctc gacttcgctg ctacccaagg
     1441 ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg aacccagtgg acataagcct
     1501 gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct
     1561 tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt
     1621 tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc
     1681 gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa
     1741 agttaaacat tatgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag
     1801 ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg
     1861 cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg gtgaccgtaa
     1921 ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc
     1981 ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca
     2041 ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca
     2101 ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa
     2161 aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg
     2221 ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc
     2281 ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg
     2341 cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc
     2401 cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag
     2461 atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg aaaggcgaga
     2521 tcaccaaggt agtcggcaaa taaccctcga gccacccatg accaaaatcc cttaacgtga
     2581 gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2641 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2701 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2761 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     2821 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2881 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2941 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3001 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3061 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3121 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3181 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3241 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3301 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3361 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     3421 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     3481 actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc caggaagagt
     3541 ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt gatgcctggc
     3601 agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc
     3661 cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg
     3721 aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag ttccctactc
     3781 tcgcgttaac gctagcatgg atgttttccc agtcacgacg tt
//
