LOCUS       sequence-329032-        4317 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 4317)
  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 4317)
  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-LCK
FEATURES             Location/Qualifiers
     source          1..4317
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(62..148)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     197..219
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     211..228
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     212..228
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    244..343
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1871..1970)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1987..2006)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1988..2006)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2011..2027)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2011..2027)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2458..3249
                     /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      3342..3930
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3831..3850
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4084..4101
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4260..4287)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg
       61 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
      121 cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg
      181 ctagcatgga tgttttccca gtcacgacgt tgtaaaacga cggccagtct taagctcggg
      241 ccccaaataa tgattttatt ttgactgata gtgacctgtt cgttgcaaca aattgatgag
      301 caatgctttt ttataatgcc aactttgtac aaaaaagttg gcaccatggg ctgtggctgc
      361 agctcacacc cggaagatga ctggatggaa aacatcgatg tgtgtgagaa ctgccattat
      421 cccatagtcc cactggatgg caagggcacg ctgctcatcc gaaatggctc tgaggtgcgg
      481 gacccactga ttacctacga aggctccaat ccgccggctt ccccactgca agacaacctg
      541 gttatcgctc tgcacagcta tgagccctct cacgacggag atctgggctt tgagaagggg
      601 gaacagctcc gcatcctgga gcagagcggc gagtggtgga aggcgcagtc cctgaccacg
      661 ggccaggaag gcttcatccc cttcaatttt gtggccaaag cgaacagcct ggagcccgaa
      721 ccctggttct tcaagaacct gagccgcaag gacgcggagc ggcagctcct ggcgcccggg
      781 aacactcacg gctccttcct catccgggag agcgagagca ccgcgggatc gttttcactg
      841 tcggtccggg acttcgacca gaaccaggga gaggtggtga aacattacaa gatccgtaat
      901 ctggacaacg gtggcttcta catctcccct cgaatcactt ttcccggcct gcatgaactg
      961 gtccgccatt acaccaatgc ttcagatggg ctgtgcacac ggttgagccg cccctgccag
     1021 acccagaagc cccagaagcc gtggtgggag gacgagtggg aggttcccag ggagacgctg
     1081 aagctggtgg agcggctggg ggctggacag ttcggggagg tgtggatggg gtactacaac
     1141 gggcacacga aggtggcggt gaagagcctg aagcagggca gcatgtcccc ggacgccttc
     1201 ctggccgagg ccaacctcat gaagcagctg caacaccagc ggctggttcg gctctacgct
     1261 gtggtcaccc aggagcccat ctacatcatc actgaataca tggagaatgg gagtctagtg
     1321 gattttctca agaccccttc aggcatcaag ttgaccatca acaaactcct ggacatggca
     1381 gcccaaattg cagaaggcat ggcattcatt gaagagcgga attatattca tcgtgacctt
     1441 cgggctgcca acattctggt gtctgacacc ctgagctgca agattgcaga ctttggccta
     1501 gcacgcctca ttgaggacaa cgagtacaca gccagggagg gggccaagtt tcccattaag
     1561 tggacagcgc cagaagccat taactacggg acattcacca tcaagtcaga tgtgtggtct
     1621 tttgggatcc tgctgacgga aattgtcacc cacggccgca tcccttaccc agggatgacc
     1681 aacccggagg tgattcagaa cctggagcga ggctaccgca tggtgcgccc tgacaactgt
     1741 ccagaggagc tgtaccaact catgaggctg tgctggaagg agcgcccaga ggaccggccc
     1801 acctttgact acctgcgcag tgtgctggag gacttcttca cggccacaga gggccagtac
     1861 cagcctcagc ctccaacttt cttgtacaaa gttggcatta taagaaagca ttgcttatca
     1921 atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct
     1981 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
     2041 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgcctcctc
     2101 tagaccagcc aggacagaaa tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc
     2161 acaccgtgga aacggatgaa ggcacgaacc cagtggacat aagcctgttc ggttcgtaag
     2221 ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc
     2281 ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt tggggtacag
     2341 tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta
     2401 tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt aaacattatg
     2461 agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg cgtcatcgag
     2521 cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt ggatggcggc
     2581 ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct tgatgaaaca
     2641 acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg agagagcgag
     2701 attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc gtggcgttat
     2761 ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct tgcaggtatc
     2821 ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc aagagaacat
     2881 agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc tgaacaggat
     2941 ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga ctgggctggc
     3001 gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa
     3061 atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc ccagtatcag
     3121 cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg cttggcctcg
     3181 cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac caaggtagtc
     3241 ggcaaataac cctcgagcca cccatgacca aaatccctta acgtgagtta cgcgtcgttc
     3301 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3361 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3421 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3481 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3541 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3601 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3661 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3721 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3781 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3841 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3901 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3961 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4021 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4081 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     4141 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     4201 agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa
     4261 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcg
//
