LOCUS       sequence-215968-        5274 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 5274)
  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 5274)
  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-KSR2
FEATURES             Location/Qualifiers
     source          1..5274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(144..171)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(263..349)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     398..420
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     412..429
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     413..429
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    445..544
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3029..3128)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3145..3164)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3146..3164)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3169..3185)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3169..3185)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3616..4407
                     /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      4500..5088
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4989..5008
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5242..5259
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
       61 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc
      121 caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt
      181 gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa
      241 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
      301 agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag
      361 ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg ttgtaaaacg
      421 acggccagtc ttaagctcgg gccccaaata atgattttat tttgactgat agtgacctgt
      481 tcgttgcaac aaattgatga gcaatgcttt tttataatgc caactttgta caaaaaagtt
      541 ggcaccatga cggatgaaca ggtgtgcgag actgtggaga aatacggagc caaccgggag
      601 gagtgtgccc gcctcaacgc ctccctctcc tgcctcagga atgtccacat gtcaggaggc
      661 aacctttcca aacaagactg gaccatccag tggcccacca cagagacggg gaaggagaac
      721 aatcccgtgt gccccccgga gcccaccccg tggatccgca cccatctctc ccagagcccc
      781 agggtcccgt ccaagtgcgt ccagcactat tgtcacacca gccccactcc cggggcccct
      841 gtgtacaccc acgtggacag gcttaccgtg gacgcctacc cgggcttgtg cccgcccccg
      901 ccactggagt cgggccaccg ttccctgccc ccatcgcccc ggcagcggca cgcggtccgc
      961 accccgccgc gcacccccaa catcgtcacc accgtgaccc cgccgggcac gccgcccatg
     1021 aggaagaaga acaagctgaa gcccccgggg accccaccgc cctcctcccg aaaactgata
     1081 cacttgatcc cgggattcac cgcgctgcat cggagcaaat cccacgagtt ccagctgggg
     1141 caccgcgtgg acgaggccca cacgcccaaa gccaagaaga agagcaaacc cttgaacctc
     1201 aagatccaca gcagcgtagg cagctgcgag aacatcccct ctcagcagcg ctccccgctg
     1261 ctgtccgagc gctccctccg ctccttcttt gtgggacacg cacctttcct gccttccacc
     1321 cctcctgttc acactgaggc caacttctct gcaaacacac tgtcagtgcc acgctggtcc
     1381 ccgcagatcc ctcgcagaga tcttggcaac tccatcaagc acaggttttc caccaagtac
     1441 tggatgtctc agacgtgcac agtctgtggg aaagggatgc tttttggcct caagtgtaaa
     1501 aactgcaagt taaagtgcca caacaaatgc accaaagaag ccccaccctg tcatcttctg
     1561 atcatccacc gaggagcaag gttagtccgg acagagtccg ttccatgtga catcaacaac
     1621 cctctacgga agccacctcg ctattcagac ctgcacatca gtcagacgct ccccaaaacc
     1681 aacaaaatca acaaggacca catccctgtc ccttaccagc cagactccag cagcaacccc
     1741 tcctccacga cgtcctccac gccctcctcg ccagcacccc ccctccctcc tagtgccacg
     1801 ccgccttctc ccctacaccc ttccccacag tgcacacggc agcagaagaa cttcaacctg
     1861 ccagcatccc actactacaa atacaagcag cagttcatct tcccagatgt ggtgccggtg
     1921 ccggagacgc cgacccgggc gccccaggtc atcctgcatc cggtgacctc gaatccaatc
     1981 ttggaaggaa atccattact tcaaattgaa gtggagccaa cgtcggagaa tgaagaggtc
     2041 catgatgagg ccgaagagtc agaggatgac ttcgaggaga tgaacctgtc cctcctctcg
     2101 gcccggggct tcccacgcaa ggccagccag accagcatct tccttcagga gtgggacatc
     2161 ccctttgagc agctggagat cggcgagctc attggaaagg gccgctttgg gcaagtgtac
     2221 cacggccgct ggcatggcga ggtggccatc cggctgattg acattgagag ggacaacgag
     2281 gaccagctca aggccttcaa gcgggaggtg atggcctaca ggcagacacg gcatgagaac
     2341 gtggtgcttt tcatgggtgc ctgcatgagc ccgcctcacc tggccatcat caccagcctc
     2401 tgtaagggac ggacgctcta ttccgttgtg agggatgcca aaatcgtttt ggatgtcaac
     2461 aaaaccaggc agattgctca agaaattgtg aagggcatgg gctacctcca cgccaaggga
     2521 atcctacaca aggacctcaa gtcaaagaac gtcttctatg acaacggcaa agtggtcatc
     2581 acggactttg gactcttcag catttctggg gtgctgcagg ctggcaggcg ggaggacaaa
     2641 ctgcgcatcc agaatggctg gctatgccac ctggcaccag agatcatccg ccagctgtcc
     2701 cccgacacag aggaggataa gctccccttc tccaagcact ctgacgtctt tgcccttggc
     2761 acaatctggt atgaactcca cgccagggaa tggcctttca agacccaacc agcagaggca
     2821 ataatctggc aaatgggcac aggcatgaaa cccaacctca gccagattgg catgggaaaa
     2881 gaaatctcgg acattcttct cttctgctgg gcctttgaac aagaagagag acctaccttc
     2941 accaagctca tggacatgct ggagaaactg ccaaagcgaa accgtcgcct gtctcaccct
     3001 ggacatttct ggaagtctgc agagctgttg ccaactttct tgtacaaagt tggcattata
     3061 agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc
     3121 attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt
     3181 tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     3241 atatcatcat gcctcctcta gaccagccag gacagaaatg cctcgacttc gctgctaccc
     3301 aaggttgccg ggtgacgcac accgtggaaa cggatgaagg cacgaaccca gtggacataa
     3361 gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga
     3421 accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac
     3481 tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg
     3541 ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa
     3601 acaaagttaa acattatgag ggaagcggtg atcgccgaag tatcgactca actatcagag
     3661 gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc
     3721 tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc
     3781 gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct
     3841 tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac
     3901 atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat
     3961 gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg
     4021 acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat
     4081 ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg
     4141 ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac
     4201 agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc
     4261 ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa
     4321 gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc
     4381 gagatcacca aggtagtcgg caaataaccc tcgagccacc catgaccaaa atcccttaac
     4441 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4501 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4561 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4621 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     4681 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4741 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4801 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4861 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4921 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4981 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5041 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5101 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5161 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     5221 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgc
//
