LOCUS       sequence-205487-        6960 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 6960)
  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 6960)
  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-MET
FEATURES             Location/Qualifiers
     source          1..6960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(54..140)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     189..211
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     203..220
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     204..220
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    236..335
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4506..4605)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4622..4641)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4623..4641)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4646..4662)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4646..4662)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5093..5884
                     /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      5977..6565
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6466..6485
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6719..6736
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6895..6922)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc
       61 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac
      121 tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg
      181 gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccccaaat
      241 aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgatg agcaatgctt
      301 ttttataatg ccaactttgt acaaaaaagt tggcaccatg aaggcccccg ctgtgcttgc
      361 acctggcatc ctcgtgctcc tgtttacctt ggtgcagagg agcaatgggg agtgtaaaga
      421 ggcactagca aagtccgaga tgaatgtgaa tatgaagtat cagcttccca acttcaccgc
      481 ggaaacaccc atccagaatg tcattctaca tgagcatcac attttccttg gtgccactaa
      541 ctacatttat gttttaaatg aggaagacct tcagaaggtt gctgagtaca agactgggcc
      601 tgtgctggaa cacccagatt gtttcccatg tcaggactgc agcagcaaag ccaatttatc
      661 aggaggtgtt tggaaagata acatcaacat ggctctagtt gtcgacacct actatgatga
      721 tcaactcatt agctgtggca gcgtcaacag agggacctgc cagcgacatg tctttcccca
      781 caatcatact gctgacatac agtcggaggt tcactgcata ttctccccac agatagaaga
      841 gcccagccag tgtcctgact gtgtggtgag cgccctggga gccaaagtcc tttcatctgt
      901 aaaggaccgg ttcatcaact tctttgtagg caataccata aattcttctt atttcccaga
      961 tcatccattg cattcgatat cagtgagaag gctaaaggaa acgaaagatg gttttatgtt
     1021 tttgacggac cagtcctaca ttgatgtttt acctgagttc agagattctt accccattaa
     1081 gtatgtccat gcctttgaaa gcaacaattt tatttacttc ttgacggtcc aaagggaaac
     1141 tctagatgct cagacttttc acacaagaat aatcaggttc tgttccataa actctggatt
     1201 gcattcctac atggaaatgc ctctggagtg tattctcaca gaaaagagaa aaaagagatc
     1261 cacaaagaag gaagtgttta atatacttca ggctgcgtat gtcagcaagc ctggggccca
     1321 gcttgctaga caaataggag ccagcctgaa tgatgacatt cttttcgggg tgttcgcaca
     1381 aagcaagcca gattctgccg aaccaatgga tcgatctgcc atgtgtgcat tccctatcaa
     1441 atatgtcaac gacttcttca acaagatcgt caacaaaaac aatgtgagat gtctccagca
     1501 tttttacgga cccaatcatg agcactgctt taataggaca cttctgagaa attcatcagg
     1561 ctgtgaagcg cgccgtgatg aatatcgaac agagtttacc acagctttgc agcgcgttga
     1621 cttattcatg ggtcaattca gcgaagtcct cttaacatct atatccacct tcattaaagg
     1681 agacctcacc atagctaatc ttgggacatc agagggtcgc ttcatgcagg ttgtggtttc
     1741 tcgatcagga ccatcaaccc ctcatgtgaa ttttctcctg gactcccatc cagtgtctcc
     1801 agaagtgatt gtggagcata cattaaacca aaatggctac acactggtta tcactgggaa
     1861 gaagatcacg aagatcccat tgaatggctt gggctgcaga catttccagt cctgcagtca
     1921 atgcctctct gccccaccct ttgttcagtg tggctggtgc cacgacaaat gtgtgcgatc
     1981 ggaggaatgc ctgagcggga catggactca acagatctgt ctgcctgcaa tctacaaggt
     2041 tttcccaaat agtgcacccc ttgaaggagg gacaaggctg accatatgtg gctgggactt
     2101 tggatttcgg aggaataata aatttgattt aaagaaaact agagttctcc ttggaaatga
     2161 gagctgcacc ttgactttaa gtgagagcac gatgaataca ttgaaatgca cagttggtcc
     2221 tgccatgaat aagcatttca atatgtccat aattatttca aatggccacg ggacaacaca
     2281 gtacagtaca ttctcctatg tggatcctgt aataacaagt atttcgccga aatacggtcc
     2341 tatggctggt ggcactttac ttactttaac tggaaattac ctaaacagtg ggaattctag
     2401 acacatttca attggtggaa aaacatgtac tttaaaaagt gtgtcaaaca atattcttga
     2461 atgttatacc ccagcccaaa ccatttcaac tgagtttgct gttaaattga aaattgactt
     2521 agccaaccga gagacaagca tcttcagtta ccgtgaagat cccattgtct atgaaattca
     2581 tccaaccaaa tcttttatta gtggtgggag cacaataaca ggtgttggga aaaacctgaa
     2641 ttcagttagt gtcccgagaa tggtcataaa tgtgcatgaa gcaggaagga actttacagt
     2701 ggcatgtcaa catcgctcta attcagagat aatctgttgt accactcctt ccctgcaaca
     2761 gctgaatctg caactccccc tgaaaaccaa agcctttttc atgttagatg ggatcctttc
     2821 caaatacttt gatctcattt atgtacataa tcctgtgttt aagccttttg aaaagccagt
     2881 gatgatctca atgggcaatg aaaatgtact ggaaattaag ggaaatgata ttgaccctga
     2941 agcagttaaa ggtgaagtgt taaaagttgg aaataagagc tgtgagaata tacacttaca
     3001 ttctgaagcc gttttatgca cggtccccaa tgacctgctg aaattgaaca gcgagctaaa
     3061 tatagagtgg aagcaagcaa tttcttcaac cgtccttgga aaagtaatag ttcaaccaga
     3121 tcagaatttc acaggattga ttgctggtgt tgtctcaata tcaacagcac tgttattact
     3181 acttgggttt ttcctgtggc tgaaaaagag aaagcaaatt aaagatctgg gcagtgaatt
     3241 agttcgctac gatgcaagag tacacactcc tcatttggat aggcttgtaa gtgcccgaag
     3301 tgtaagccca actacagaaa tggtttcaaa tgaatctgta gactaccgag ctacttttcc
     3361 agaagatcag tttcctaatt catctcagaa cggttcatgc cgacaagtgc agtatcctct
     3421 gacagacatg tcccccatcc taactagtgg ggactctgat atatccagtc cattactgca
     3481 aaatactgtc cacattgacc tcagtgctct aaatccagag ctggtccagg cagtgcagca
     3541 tgtagtgatt gggcccagta gcctgattgt gcatttcaat gaagtcatag gaagagggca
     3601 ttttggttgt gtatatcatg ggactttgtt ggacaatgat ggcaagaaaa ttcactgtgc
     3661 tgtgaaatcc ttgaacagaa tcactgacat aggagaagtt tcccaatttc tgaccgaggg
     3721 aatcatcatg aaagatttta gtcatcccaa tgtcctctcg ctcctgggaa tctgcctgcg
     3781 aagtgaaggg tctccgctgg tggtcctacc atacatgaaa catggagatc ttcgaaattt
     3841 cattcgaaat gagactcata atccaactgt aaaagatctt attggctttg gtcttcaagt
     3901 agccaaaggc atgaaatatc ttgcaagcaa aaagtttgtc cacagagact tggctgcaag
     3961 aaactgtatg ctggatgaaa aattcacagt caaggttgct gattttggtc ttgccagaga
     4021 catgtatgat aaagaatact atagtgtaca caacaaaaca ggtgcaaagc tgccagtgaa
     4081 gtggatggct ttggaaagtc tgcaaactca aaagtttacc accaagtcag atgtgtggtc
     4141 ctttggcgtg ctcctctggg agctgatgac aagaggagcc ccaccttatc ctgacgtaaa
     4201 cacctttgat ataactgttt acttgttgca agggagaaga ctcctacaac ccgaatactg
     4261 cccagacccc ttatatgaag taatgctaaa atgctggcac cctaaagccg aaatgcgccc
     4321 atccttttct gaactggtgt cccggatatc agcgatcttc tctactttca ttggggagca
     4381 ctatgtccat gtgaacgcta cttatgtgaa cgtaaaatgt gtcgctccgt atccttctct
     4441 gttgtcatca gaagataacg ctgatgatga ggtggacaca cgaccagcct ccttctggga
     4501 gacatcacca actttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     4561 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     4621 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     4681 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgcc tcctctagac
     4741 cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc
     4801 gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta
     4861 atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg
     4921 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat
     4981 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
     5041 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga
     5101 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca
     5161 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa
     5221 gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg
     5281 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct
     5341 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc
     5401 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga
     5461 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt
     5521 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt
     5581 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga
     5641 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc
     5701 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt
     5761 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc
     5821 agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa
     5881 ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     5941 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6001 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6061 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6121 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6181 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6241 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6301 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6361 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6421 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6481 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6541 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6601 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6661 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     6721 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     6781 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     6841 gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg
     6901 ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct
//
