LOCUS       sequence-220461-        6861 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 6861)
  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 6861)
  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-KDR
FEATURES             Location/Qualifiers
     source          1..6861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(23..109)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     158..180
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     172..189
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     173..189
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    205..304
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4376..4475)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4492..4511)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4493..4511)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4516..4532)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4516..4532)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4963..5754
                     /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      5847..6435
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6336..6355
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6589..6606
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6765..6792)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
       61 agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag
      121 ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg ttgtaaaacg
      181 acggccagtc ttaagctcgg gccccaaata atgattttat tttgactgat agtgacctgt
      241 tcgttgcaac aaattgatga gcaatgcttt tttataatgc caactttgta caaaaaagtt
      301 ggcaccatgc agagcaaggt gctgctggcc gtcgccctgt ggctctgcgt ggagacccgg
      361 gccgcctctg tgggtttgcc tagtgtttct cttgatctgc ccaggctcag catacaaaaa
      421 gacatactta caattaaggc taatacaact cttcaaatta cttgcagggg acagagggac
      481 ttggactggc tttggcccaa taatcagagt ggcagtgagc aaagggtgga ggtgactgag
      541 tgcagcgatg gcctcttctg taagacactc acaattccaa aagtgatcgg aaatgacact
      601 ggagcctaca agtgcttcta ccgggaaact gacttggcct cggtcattta tgtctatgtt
      661 caagattaca gatctccatt tattgcttct gttagtgacc aacatggagt cgtgtacatt
      721 actgagaaca aaaacaaaac tgtggtgatt ccatgtctcg ggtccatttc aaatctcaac
      781 gtgtcacttt gtgcaagata cccagaaaag agatttgttc ctgatggtaa cagaatttcc
      841 tgggacagca agaagggctt tactattccc agctacatga tcagctatgc tggcatggtc
      901 ttctgtgaag caaaaattaa tgatgaaagt taccagtcta ttatgtacat agttgtcgtt
      961 gtagggtata ggatttatga tgtggttctg agtccgtctc atggaattga actatctgtt
     1021 ggagaaaagc ttgtcttaaa ttgtacagca agaactgaac taaatgtggg gattgacttc
     1081 aactgggaat acccttcttc gaagcatcag cataagaaac ttgtaaaccg agacctaaaa
     1141 acccagtctg ggagtgagat gaagaaattt ttgagcacct taactataga tggtataacc
     1201 cggagtgacc aaggattgta cacctgtgca gcatccagtg ggctgatgac caagaagaac
     1261 agcacatttg tcagggtcca tgaaaaacct tttgttgctt ttggaagtgg catggaatct
     1321 ctggtggaag ccacggtggg ggagcgtgtc agaatccctg cgaagtacct tggttaccca
     1381 cccccagaaa taaaatggta taaaaatgga ataccccttg agtccaatca cacaattaaa
     1441 gcggggcatg tactgacgat tatggaagtg agtgaaagag acacaggaaa ttacactgtc
     1501 atccttacca atcccatttc aaaggagaag cagagccatg tggtctctct ggttgtgtat
     1561 gtcccacccc agattggtga gaaatctcta atctctcctg tggattccta ccagtacggc
     1621 accactcaaa cgctgacatg tacggtctat gccattcctc ccccgcatca catccactgg
     1681 tattggcagt tggaggaaga gtgcgccaac gagcccagcc aagctgtctc agtgacaaac
     1741 ccataccctt gtgaagaatg gagaagtgtg gaggacttcc agggaggaaa taaaattgaa
     1801 gttaataaaa atcaatttgc tctaattgaa ggaaaaaaca aaactgtaag tacccttgtt
     1861 atccaagcgg caaatgtgtc agctttgtac aaatgtgaag cggtcaacaa agtcgggaga
     1921 ggagagaggg tgatctcctt ccacgtgacc aggggtcctg aaattacttt gcaacctgac
     1981 atgcagccca ctgagcagga gagcgtgtct ttgtggtgca ctgcagacag atctacgttt
     2041 gagaacctca catggtacaa gcttggccca cagcctctgc caatccatgt gggagagttg
     2101 cccacacctg tttgcaagaa cttggatact ctttggaaat tgaatgccac catgttctct
     2161 aatagcacaa atgacatttt gatcatggag cttaagaatg catccttgca ggaccaagga
     2221 gactatgtct gccttgctca agacaggaag accaagaaaa gacattgcgt ggtcaggcag
     2281 ctcacagtcc tagagcgtgt ggcacccacg atcacaggaa acctggagaa tcagacgaca
     2341 agtattgggg aaagcatcga agtctcatgc acggcatctg ggaatccccc tccacagatc
     2401 atgtggttta aagataatga gacccttgta gaagactcag gcattgtatt gaaggatggg
     2461 aaccggaacc tcactatccg cagagtgagg aaggaggacg aaggcctcta cacctgccag
     2521 gcatgcagtg ttcttggctg tgcaaaagtg gaggcatttt tcataataga aggtgcccag
     2581 gaaaagacga acttggaaat cattattcta gtaggcacgg cggtgattgc catgttcttc
     2641 tggctacttc ttgtcatcat cctacggacc gttaagcggg ccaatggagg ggaactgaag
     2701 acaggctact tgtccatcgt catggatcca gatgaactcc cattggatga acattgtgaa
     2761 cgactgcctt atgatgccag caaatgggaa ttccccagag accggctgaa gctaggtaag
     2821 cctcttggcc gtggtgcctt tggccaagtg attgaagcag atgcctttgg aattgacaag
     2881 acagcaactt gcaggacagt agcagtcaaa atgttgaaag aaggagcaac acacagtgag
     2941 catcgagctc tcatgtctga actcaagatc ctcattcata ttggtcacca tctcaatgtg
     3001 gtcaaccttc taggtgcctg taccaagcca ggagggccac tcatggtgat tgtggaattc
     3061 tgcaaatttg gaaacctgtc cacttacctg aggagcaaga gaaatgaatt tgtcccctac
     3121 aagaccaaag gggcacgatt ccgtcaaggg aaagactacg ttggagcaat ccctgtggat
     3181 ctgaaacggc gcttggacag catcaccagt agccagagct cagccagctc tggatttgtg
     3241 gaggagaagt ccctcagtga tgtagaagaa gaggaagctc ctgaagatct gtataaggac
     3301 ttcctgacct tggagcatct catctgttac agcttccaag tggctaaggg catggagttc
     3361 ttggcatcgc gaaagtgtat ccacagggac ctggcggcac gaaatatcct cttatcggag
     3421 aagaacgtgg ttaaaatctg tgactttggc ttggcccggg atatttataa agatccagat
     3481 tatgtcagaa aaggagatgc tcgcctccct ttgaaatgga tggccccaga aacaattttt
     3541 gacagagtgt acacaatcca gagtgacgtc tggtcttttg gtgttttgct gtgggaaata
     3601 ttttccttag gtgcttctcc atatcctggg gtaaagattg atgaagaatt ttgtaggcga
     3661 ttgaaagaag gaactagaat gagggcccct gattatacta caccagaaat gtaccagacc
     3721 atgctggact gctggcacgg ggagcccagt cagagaccca cgttttcaga gttggtggaa
     3781 catttgggaa atctcttgca agctaatgct cagcaggatg gcaaagacta cattgttctt
     3841 ccgatatcag agactttgag catggaagag gattctggac tctctctgcc tacctcacct
     3901 gtttcctgta tggaggagga ggaagtatgt gaccccaaat tccattatga caacacagca
     3961 ggaatcagtc agtatctgca gaacagtaag cgaaagagcc ggcctgtgag tgtaaaaaca
     4021 tttgaagata tcccgttaga agaaccagaa gtaaaagtaa tcccagatga caaccagacg
     4081 gacagtggta tggttcttgc ctcagaagag ctgaaaactt tggaagacag aaccaaatta
     4141 tctccatctt ttggtggaat ggtgcccagc aaaagcaggg agtctgtggc atctgaaggc
     4201 tcaaaccaga caagcggcta ccagtccgga tatcactccg atgacacaga caccaccgtg
     4261 tactccagtg aggaagcaga acttttaaag ctgatagaga ttggagtgca aaccggtagc
     4321 acagcccaga ttctccagcc tgactcgggg accacactga gctctcctcc tgttttgcca
     4381 actttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa
     4441 caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt
     4501 gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc
     4561 tgatgttaca ttgcacaaga taaaaatata tcatcatgcc tcctctagac cagccaggac
     4621 agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg
     4681 atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag
     4741 cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca
     4801 gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca
     4861 tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat
     4921 gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc
     4981 gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg
     5041 acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt
     5101 gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg
     5161 atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta
     5221 gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa
     5281 ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg
     5341 atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta
     5401 ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta
     5461 aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta
     5521 gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat
     5581 gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa
     5641 gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg
     5701 gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg
     5761 agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac
     5821 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5881 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5941 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6001 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6061 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6121 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6181 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6241 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6301 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6361 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6421 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6481 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6541 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     6601 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     6661 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     6721 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca
     6781 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
     6841 ctccgggccg ttgcttcaca a
//
