LOCUS       sequence-132728-        6972 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 6972)
  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 6972)
  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-LRPPRC
FEATURES             Location/Qualifiers
     source          1..6972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(149..176)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(268..354)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     403..425
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     417..434
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     418..434
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    450..549
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4732..4831)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4848..4867)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4849..4867)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4872..4888)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4872..4888)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5319..6110
                     /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      6203..6791
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6692..6711
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6945..6962
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
      121 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
      181 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
      241 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
      301 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
      361 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
      421 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
      481 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
      541 aagttggcac catggcagcc ctgctgagat ccgcgcgttg gttgctgcgt gccggggcgg
      601 ccccgcgcct cccgctctcc ctgcgcctcc tccctggcgg cccgggccgg ctgcatgccg
      661 cctcctatct gcccgccgct cgcgccgggc ccgtggccgg aggactactg agcccagcca
      721 ggctgtatgc cattgctgcc aaagaaaaag atattcaaga ggagtccact ttttcttcta
      781 ggaagatttc caatcaattt gattgggctc taatgagact agatctttct gttcgaagaa
      841 ctggccgcat tccaaagaag cttctacaaa aagtttttaa tgatacctgc cgctcaggtg
      901 gcctaggtgg tagtcatgcc ttgcttctac tacgtagttg tggttctctc ttgcctgaac
      961 taaagcttga agagagaaca gaatttgctc ataggatatg ggacacactt cagaaattag
     1021 gtgctgtgta tgatgtgagt cactataatg ctttacttaa agtctatctt caaaatgaat
     1081 ataaattctc accaactgat ttcctggcaa aaatggagga agcaaacatt caaccaaatc
     1141 gagtgacata ccagagattg attgcttctt attgtaatgt aggagatatt gaaggtgcca
     1201 gcaagattct tggatttatg aaaactaagg atctcccagt tacagaggca gtattcagtg
     1261 cccttgtgac agggcatgcc agagctggtg atatggagaa tgcagaaaac attctcacag
     1321 tgatgagaga tgccggaatt gagcctggtc cagacacata cctcgcatta ttgaatgcat
     1381 atgctgagaa gggcgacatt gaccatgtta agcagactct ggagaaggtg gagaagtccg
     1441 agcttcacct tatggaccgt gatttactgc aaattatttt tagcttcagt aaagctgggt
     1501 atcctcagta tgtctcagaa attttggaaa aagttacatg tgaaagaaga tatattccag
     1561 atgcaatgaa cctcatttta cttttagtca ctgaaaaatt ggaagatgta gcgttgcaaa
     1621 ttttactagc atgccccgta tcaaaggaag atggcccaag tgtctttggc agtttctttt
     1681 tacaacactg tgtgactatg aatacgcctg tggagaagct aacagactac tgtaagaagt
     1741 taaaggaagt ccagatgcac tcctttcctc tgcagttcac cctccattgt gctttactcg
     1801 ccaataaaac tgatttggca aaagccttaa tgaaggctgt gaaggaggaa ggttttccta
     1861 tcagacctca ctatttctgg ccattgctag ttggacgtcg gaaggaaaaa aatgttcaag
     1921 gtataattga aatcctcaaa ggaatgcaag aattgggagt acatcctgat caggaaacat
     1981 atacagatta tgtgattcca tgctttgata gtgtaaactc agcacgagcc attttgcagg
     2041 aaaatggatg tctgtctgat agtgatatgt tttctcaagc tggattgaga agtgaagcag
     2101 caaatgggaa cttagacttt gtattatcat ttttgaaatc aaatacattg cccatctcgc
     2161 tgcagtctat aagaagtagc ctactgctag gcttcaggag gtctatgaat ataaatcttt
     2221 ggagcgagat aacagaattg ttgtacaagg atggacgtta ttgccaggag cctcgaggac
     2281 cgacggctgt tggctatttt ctttataact tgattgacag catgagtgac tcagaggtac
     2341 aggccaagga ggagcatttg agacaatact tccatcagct ggagaagatg aatgtaaaaa
     2401 ttcctgaaaa tatctacaga ggcattcgta atctcctgga aagctaccat gttcctgaat
     2461 tgattaagga tgctcacttg ttggttgaga gtaagaattt agactttcaa aaaactgtgc
     2521 aacttacatc atctgaattg gagtccacac ttgaaacact aaaagctgaa aatcaaccta
     2581 taagagatgt cctaaagcaa ctcatattag tgctttgttc agaagagaat atgcaaaaag
     2641 cccttgaatt gaaagcaaaa tatgaatccg acatggttac tggtggctat gcagctttaa
     2701 taaatttatg ctgtcgacat gataaagtag aagatgcctt gaacttgaaa gaagaatttg
     2761 accgcttaga ttcatctgct gtccttgaca ccggcaagta tgtaggcctt gtaagagtat
     2821 tggcaaagca tggcaagctc caagatgcta ttaacattct gaaggagatg aaagagaagg
     2881 atgttcttat caaagataca acagccttgt cctttttcca catgctaaat ggcgcagctt
     2941 taagaggtga aattgaaaca gtaaaacagt tgcatgaagc catcgtgact ctagggttag
     3001 cagaaccatc caccaacata agtttcccat tggtcactgt acacttggaa aagggcgacc
     3061 tatctactgc tcttgaggtc gccattgact gctatgaaaa gtataaagta ttaccaagga
     3121 ttcatgatgt cttgtgtaaa ctggtagaga aaggcgagac tgatctaatt cagaaagcaa
     3181 tggactttgt gagccaagaa caaggtgaaa tggtgatgct ctatgatctc ttctttgcct
     3241 tcctacaaac aggaaattac aaagaggcca agaagatcat tgagactcca gggattagag
     3301 ctcgatctgc aaggcttcag tggttttgtg acagatgtgt tgcaaataat caggttgaaa
     3361 ctctggaaaa attagtggag ctgacacaga agctatttga atgtgataga gaccagatgt
     3421 actacaatct gctaaaactg tataaaataa acggtgactg gcaaagagct gatgcagtct
     3481 ggaataaaat ccaagaagaa aatgttattc ctcgtgaaaa gacattaaga ttattagcag
     3541 aaatccttag agagggtaac caggaagttc cgtttgacgt acctgagttg tggtatgaag
     3601 atgaaaaaca ttccctgaat tcttcgtcag cctcaaccac agaacctgat ttccagaaag
     3661 atatattgat tgcctgccga ttgaaccaaa aaaaaggggc atatgatatt ttcctgaatg
     3721 caaaagagca aaacattgtg tttaatgctg aaacctacag caatctcatt aaattactga
     3781 tgtcagaaga ttattttaca caagcaatgg aagtgaaagc attcgcggag acccacatca
     3841 agggcttcac actgaacgat gctgccaaca gccgcctcat cataacgcaa gttaggcggg
     3901 attatttgaa agaggctgtg acaacactga aaacagtatt ggatcagcag cagacccctt
     3961 ctaggttagc agtgacccgt gtcatccagg cattggccat gaagggtgat gttgaaaaca
     4021 tagaagtagt tcagaagatg ttaaatggac tcgaagactc cattggactt tcaaaaatgg
     4081 ttttcatcaa taacattgct ttggctcaaa taaagaataa taacatagat gccgcaatag
     4141 aaaacattga aaatatgctt acttcagaga ataaagtcat tgaaccccaa tacttcggct
     4201 tggcatactt attcagaaaa gtaatagagg agcagttgga accagcagtt gaaaagataa
     4261 gcatcatggc ggagagattg gccaatcagt ttgcaattta taaacctgtc actgattttt
     4321 tccttcaact tgtggatgca ggcaaggtgg atgatgccag agctctccta cagagatgtg
     4381 gtgcaattgc tgaacaaacc ccgattttgt tgttgttcct ccttaggaat tctaggaaac
     4441 aaggaaaggc atcaactgtg aaatctgtgt tagaattgat tcctgaatta aatgaaaagg
     4501 aagaagcata caattccctc atgaaaagct atgtctcaga gaaagatgtc acatctgcta
     4561 aagcactgta tgaacatttg actgcaaaga atacaaaatt ggatgatctg tttctaaagc
     4621 gttacgcatc tttgctgaag tatgctggag agcctgtccc tttcattgaa ccccctgaaa
     4681 gctttgaatt ttatgcacag cagctaagaa aattgaggga aaactcttct ttgccaactt
     4741 tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg
     4801 tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt
     4861 cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat
     4921 gttacattgc acaagataaa aatatatcat catgcctcct ctagaccagc caggacagaa
     4981 atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg aaacggatga
     5041 aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta
     5101 tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg
     5161 cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca
     5221 agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta
     5281 cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg gtgatcgccg
     5341 aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt
     5401 tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata
     5461 ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca
     5521 acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag
     5581 tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc
     5641 aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg
     5701 acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc
     5761 cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg
     5821 aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc
     5881 ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg
     5941 ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta
     6001 gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag
     6061 aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc
     6121 acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg
     6181 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6241 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6301 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6361 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6421 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6481 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6541 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6601 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6661 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6721 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6781 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6841 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     6901 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6961 aggaagcgga ag
//
