LOCUS       sequence-263320-        6006 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 6006)
  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 6006)
  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-RET
FEATURES             Location/Qualifiers
     source          1..6006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     32..51
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     285..302
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(461..488)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(580..666)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     715..737
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     729..746
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     730..746
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    762..861
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4078..4177)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4194..4213)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4195..4213)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4218..4234)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4218..4234)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4665..5456
                     /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      join(5549..6006,1..131)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
       61 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
      121 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
      181 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
      241 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
      301 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
      361 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
      421 atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat
      481 ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc
      541 gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc
      601 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt
      661 tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg ttttcccagt
      721 cacgacgttg taaaacgacg gccagtctta agctcgggcc ccaaataatg attttatttt
      781 gactgatagt gacctgttcg ttgcaacaaa ttgatgagca atgctttttt ataatgccaa
      841 ctttgtacaa aaaagttggc accatggcga aggcgacgtc cggtgccgcg gggctgcgtc
      901 tgctgttgct gctgctgctg ccgctgctag gcaaagtggc attgggcctc tacttctcga
      961 gggatgctta ctgggagaag ctgtatgtgg accaggcggc cggcacgccc ttgctgtacg
     1021 tccatgccct gcgggacgcc cctgaggagg tgcccagctt ccgcctgggc cagcatctct
     1081 acggcacgta ccgcacacgg ctgcatgaga acaactggat ctgcatccag gaggacaccg
     1141 gcctcctcta ccttaaccgg agcctggacc atagctcctg ggagaagctc agtgtccgca
     1201 accgcggctt tcccctgctc accgtctacc tcaaggtctt cctgtcaccc acatcccttc
     1261 gtgagggcga gtgccagtgg ccaggctgtg cccgcgtata cttctccttc ttcaacacct
     1321 cctttccagc ctgcagctcc ctcaagcccc gggagctctg cttcccagag acaaggccct
     1381 ccttccgcat tcgggagaac cgacccccag gcaccttcca ccagttccgc ctgctgcctg
     1441 tgcagttctt gtgccccaac atcagcgtgg cctacaggct cctggagggt gagggtctgc
     1501 ccttccgctg cgccccggac agcctggagg tgagcacgcg ctgggccctg gaccgcgagc
     1561 agcgggagaa gtacgagctg gtggccgtgt gcaccgtgca cgccggcgcg cgcgaggagg
     1621 tggtgatggt gcccttcccg gtgaccgtgt acgacgagga cgactcggcg cccaccttcc
     1681 ccgcgggcgt cgacaccgcc agcgccgtgg tggagttcaa gcggaaggag gacaccgtgg
     1741 tggccacgct gcgtgtcttc gatgcagacg tggtacctgc atcaggggag ctggtgaggc
     1801 ggtacacaag cacgctgctc cccggggaca cctgggccca gcagaccttc cgggtggaac
     1861 actggcccaa cgagacctcg gtccaggcca acggcagctt cgtgcgggcg accgtacatg
     1921 actataggct ggttctcaac cggaacctct ccatctcgga gaaccgcacc atgcagctgg
     1981 cggtgctggt caatgactca gacttccagg gcccaggagc gggcgtcctc ttgctccact
     2041 tcaacgtgtc ggtgctgccg gtcagcctgc acctgcccag tacctactcc ctctccgtga
     2101 gcaggagggc tcgccgattt gcccagatcg ggaaagtctg tgtggaaaac tgccaggcat
     2161 tcagtggcat caacgtccag tacaagctgc attcctctgg tgccaactgc agcacgctag
     2221 gggtggtcac ctcagccgag gacacctcgg ggatcctgtt tgtgaatgac accaaggccc
     2281 tgcggcggcc caagtgtgcc gaacttcact acatggtggt ggccaccgac cagcagacct
     2341 ctaggcaggc ccaggcccag ctgcttgtaa cagtggaggg gtcatatgtg gccgaggagg
     2401 cgggctgccc cctgtcctgt gcagtcagca agagacggct ggagtgtgag gagtgtggcg
     2461 gcctgggctc cccaacaggc aggtgtgagt ggaggcaagg agatggcaaa gggatcacca
     2521 ggaacttctc cacctgctct cccagcacca agacctgccc cgacggccac tgcgatgttg
     2581 tggagaccca agacatcaac atttgccctc aggactgcct ccggggcagc attgttgggg
     2641 gacacgagcc tggggagccc cgggggatta aagctggcta tggcacctgc aactgcttcc
     2701 ctgaggagga gaagtgcttc tgcgagcccg aagacatcca ggatccactg tgcgacgagc
     2761 tgtgccgcac ggtgatcgca gccgctgtcc tcttctcctt catcgtctcg gtgctgctgt
     2821 ctgccttctg catccactgc taccacaagt ttgcccacaa gccacccatc tcctcagctg
     2881 agatgacctt ccggaggccc gcccaggcct tcccggtcag ctactcctct tccagtgccc
     2941 gccggccctc gctggactcc atggagaacc aggtctccgt ggatgccttc aagatcctgg
     3001 aggatccaaa gtgggaattc cctcggaaga acttggttct tggaaaaact ctaggagaag
     3061 gcgaatttgg aaaagtggtc aaggcaacgg ccttccatct gaaaggcaga gcagggtaca
     3121 ccacggtggc cgtgaagatg ctgaaagaga acgcctcccc gagtgagctt cgagacctgc
     3181 tgtcagagtt caacgtcctg aagcaggtca accacccaca tgtcatcaaa ttgtatgggg
     3241 cctgcagcca ggatggcccg ctcctcctca tcgtggagta cgccaaatac ggctccctgc
     3301 ggggcttcct ccgcgagagc cgcaaagtgg ggcctggcta cctgggcagt ggaggcagcc
     3361 gcaactccag ctccctggac cacccggatg agcgggccct caccatgggc gacctcatct
     3421 catttgcctg gcagatctca caggggatgc agtatctggc cgagatgaag ctcgttcatc
     3481 gggacttggc agccagaaac atcctggtag ctgaggggcg gaagatgaag atttcggatt
     3541 tcggcttgtc ccgagatgtt tatgaagagg attcgtacgt gaagaggagc cagggtcgga
     3601 ttccagttaa atggatggca attgaatccc tttttgatca tatctacacc acgcaaagtg
     3661 atgtatggtc ttttggtgtc ctgctgtggg agatcgtgac cctaggggga aacccctatc
     3721 ctgggattcc tcctgagcgg ctcttcaacc ttctgaagac cggccaccgg atggagaggc
     3781 cagacaactg cagcgaggag atgtactgcc tgatgctgca atgctggaag caggagccgg
     3841 acaaaaggcc ggtgtttgcg gacatcagca aagacctgga gaagatgatg gttaagagga
     3901 gagactactt ggaccttgcg gcgtccactc catctgactc cctgatttat gacgacggcc
     3961 tctcagagga ggagacaccg ctggtggact gtaataatgc ccccctccct cgagccctcc
     4021 cttccacatg gattgaaaac aaactctatg gtagaatttc ccatgcattt actagattcc
     4081 caactttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg
     4141 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata
     4201 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
     4261 tctgatgtta cattgcacaa gataaaaata tatcatcatg cctcctctag accagccagg
     4321 acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac
     4381 ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt
     4441 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg
     4501 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg
     4561 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
     4621 atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga
     4681 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
     4741 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
     4801 gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
     4861 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
     4921 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
     4981 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
     5041 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
     5101 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
     5161 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
     5221 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
     5281 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
     5341 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
     5401 tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct
     5461 cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     5521 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5581 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5641 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5701 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5761 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5821 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5881 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5941 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6001 gggtcg
//
