LOCUS       sequence-344988-        6993 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 6993)
  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 6993)
  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-MST1R
FEATURES             Location/Qualifiers
     source          1..6993
                     /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"
     protein_bind    complement(4336..4435)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4452..4471)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4453..4471)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4476..4492)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4476..4492)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4923..5714
                     /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      5807..6395
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6296..6315
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6549..6566
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6725..6752)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6844..6930)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(6979..6993,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6993,1..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 caccatggag ctcctcccgc cgctgcctca gtccttcctg ttgctgctgc
      181 tgttgcctgc caagcccgcg gcgggcgagg actggcagtg cccgcgcacc ccctacgcgg
      241 cctctcgcga ctttgacgtg aagtacgtgg tgcccagctt ctccgccgga ggcctggtac
      301 aggccatggt gacctacgag ggcgacagaa atgagagtgc tgtgtttgta gccatacgca
      361 atcgcctgca tgtgcttggg cctgacctga agtctgtcca gagcctggcc acgggccctg
      421 ctggagaccc tggctgccag acgtgtgcag cctgtggccc aggaccccac ggccctcccg
      481 gtgacacaga cacaaaggtg ctggtgctgg atcccgcgct gcctgcgctg gtcagttgtg
      541 gctccagcct gcagggccgc tgcttcctgc atgacctaga gccccaaggg acagccgtgc
      601 atctggcagc gccagcctgc ctcttctcag cccaccataa ccggcccgat gactgccccg
      661 actgtgtggc cagcccattg ggcacccgtg taactgtggt tgagcaaggc caggcctcct
      721 atttctacgt ggcatcctca ctggacgcag ccgtggctgc cagcttcagc ccacgctcag
      781 tgtctatcag gcgtctcaag gctgacgcct cgggattcgc accgggcttt gtggcgttgt
      841 cagtgctgcc caagcatctt gtctcctaca gtattgaata cgtgcacagc ttccacacgg
      901 gagccttcgt atacttcctg actgtacagc cggccagcgt gacagatgat cctagtgccc
      961 tgcacacacg cctggcacgg cttagcgcca ctgagccaga gttgggtgac tatcgggagc
     1021 tggtcctcga ctgcagattt gctccaaaac gcaggcgccg gggggcccca gaaggcggac
     1081 agccctaccc tgtgctgcag gtggcccact ccgctccagt gggtgcccaa cttgccactg
     1141 agctgagcat cgccgagggc caggaagtac tatttggggt ctttgtgact ggcaaggatg
     1201 gtggtcctgg cgtgggcccc aactctgtcg tctgtgcctt ccccattgac ctgctggaca
     1261 cactaattga tgagggtgtg gagcgctgtt gtgaatcccc agtccatcca ggcctccggc
     1321 gaggcctcga cttcttccag tcgcccagtt tttgccccaa cccgcctggc ctggaagccc
     1381 tcagccccaa caccagctgc cgccacttcc ctctgctggt cagtagcagc ttctcacgtg
     1441 tggacctatt caatgggctg ttgggaccag tacaggtcac tgcattgtat gtgacacgcc
     1501 ttgacaacgt cacagtggca cacatgggca caatggatgg gcgtatcctg caggtggagc
     1561 tggtcaggtc actaaactac ttgctgtatg tgtccaactt ctcactgggt gacagtgggc
     1621 agcccgtgca gcgggatgtc agtcgtcttg gggaccacct actctttgcc tctggggacc
     1681 aggttttcca ggtacctatc caaggccctg gctgccgcca cttcctgacc tgtgggcgtt
     1741 gcctaagggc atggcatttc atgggctgtg gctggtgtgg gaacatgtgc ggccagcaga
     1801 aggagtgtcc tggctcctgg caacaggacc actgcccacc taagcttact gagttccacc
     1861 cccacagtgg acctctaagg ggcagtacaa ggctgaccct gtgtggctcc aacttctacc
     1921 ttcacccttc tggtctggtg cctgagggaa cccatcaggt cactgtgggc caaagtccct
     1981 gccggccact gcccaaggac agctcaaaac tcagaccagt gccccggaaa gactttgtag
     2041 aggagtttga gtgtgaactg gagcccttgg gcacccaggc agtggggcct accaacgtca
     2101 gcctcaccgt gactaacatg ccaccgggca agcacttccg ggtagacggc acctccgtgc
     2161 tgagaggctt ctctttcatg gagccagtgc tgatagcagt gcaacccctc tttggcccac
     2221 gggcaggagg cacctgtctc actcttgaag gccagagtct gtctgtaggc accagccggg
     2281 ctgtgctggt caatgggact gagtgtctgc tagcacgggt cagtgagggg cagcttttat
     2341 gtgccacacc ccctggggcc acggtggcca gtgtccccct tagcctgcag gtggggggtg
     2401 cccaggtacc tggttcctgg accttccagt acagagaaga ccctgtcgtg ctaagcatca
     2461 gccccaactg tggctacatc aactcccaca tcaccatctg tggccagcat ctaacttcag
     2521 catggcactt agtgctgtca ttccatgacg ggcttagggc agtggaaagc aggtgtgaga
     2581 ggcagcttcc agagcagcag ctgtgccgcc ttcctgaata tgtggtccga gacccccagg
     2641 gatgggtggc agggaatctg agtgcccgag gggatggagc tgctggcttt acactgcctg
     2701 gctttcgctt cctaccccca ccccatccac ccagtgccaa cctagttcca ctgaagcctg
     2761 aggagcatgc cattaagttt gagtatattg ggctgggcgc tgtggctgac tgtgtgggta
     2821 tcaacgtgac cgtgggtggt gagagctgcc agcacgagtt ccggggggac atggttgtct
     2881 gccccctgcc cccatccctg cagcttggcc aggatggtgc cccattgcag gtctgcgtag
     2941 atggtgaatg tcatatcctg ggtagagtgg tgcggccagg gccagatggg gtcccacaga
     3001 gcacgctcct tggtatcctg ctgcctttgc tgctgcttgt ggctgcactg gcgactgcac
     3061 tggtcttcag ctactggtgg cggaggaagc agctagttct tcctcccaac ctgaatgacc
     3121 tggcatccct ggaccagact gctggagcca cacccctgcc tattctgtac tcgggctctg
     3181 actacagaag tggccttgca ctccctgcca ttgatggtct ggattccacc acttgtgtcc
     3241 atggagcatc cttctccgat agtgaagatg aatcctgtgt gccactgctg cggaaagagt
     3301 ccatccagct aagggacctg gactctgcgc tcttggctga ggtcaaggat gtgctgattc
     3361 cccatgagcg ggtggtcacc cacagtgacc gagtcattgg caaaggccac tttggagttg
     3421 tctaccacgg agaatacata gaccaggccc agaatcgaat ccaatgtgcc atcaagtcac
     3481 taagtcgcat cacagagatg cagcaggtgg aggccttcct gcgagagggg ctgctcatgc
     3541 gtggcctgaa ccacccgaat gtgctggctc tcattggtat catgttgcca cctgagggcc
     3601 tgccccatgt gctgctgccc tatatgtgcc acggtgacct gctccagttc atccgctcac
     3661 ctcagcggaa ccccaccgtg aaggacctca tcagctttgg cctgcaggta gcccgcggca
     3721 tggagtacct ggcagagcag aagtttgtgc acagggacct ggctgcgcgg aactgcatgc
     3781 tggacgagtc attcacagtc aaggtggctg actttggttt ggcccgcgac atcctggaca
     3841 gggagtacta tagtgttcaa cagcatcgcc acgctcgcct acctgtgaag tggatggcgc
     3901 tggagagcct gcagacctat agatttacca ccaagtctga tgtgtggtca tttggtgtgc
     3961 tgctgtggga actgctgaca cggggtgccc caccataccg ccacattgac ccttttgacc
     4021 ttacccactt cctggcccag ggtcggcgcc tgccccagcc tgagtattgc cctgattctc
     4081 tgtaccaagt gatgcagcaa tgctgggagg cagacccagc agtgcgaccc accttcagag
     4141 tactagtggg ggaggtggag cagatagtgt ctgcactgct tggggaccat tatgtgcagc
     4201 tgccagcaac ctacatgaac ttgggcccca gcacctcgca tgagatgaat gtgcgtccag
     4261 aacagccgca gttctcaccc atgccaggga atgtacgccg gccccggcca ctctcagagc
     4321 ctcctcggcc cacttagcca actttcttgt acaaagttgg cattataaga aagcattgct
     4381 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc
     4441 cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
     4501 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgcc
     4561 tcctctagac cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt
     4621 gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc
     4681 gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
     4741 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg
     4801 tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga
     4861 tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca
     4921 ttatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca
     4981 tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg
     5041 gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg
     5101 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga
     5161 gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc
     5221 gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag
     5281 gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag
     5341 aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac
     5401 aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg
     5461 ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg
     5521 gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt
     5581 atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg
     5641 cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg
     5701 tagtcggcaa ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt
     5761 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5821 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5881 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5941 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6001 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6061 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6121 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6181 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6241 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6301 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6361 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6421 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6481 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     6541 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     6601 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     6661 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
     6721 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
     6781 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
     6841 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     6901 gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa
     6961 cgctagcatg gatgttttcc cagtcacgac gtt
//
