LOCUS       sequence-181018-        6057 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 6057)
  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 6057)
  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-PDGFRA
FEATURES             Location/Qualifiers
     source          1..6057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(147..174)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(266..352)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     401..423
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     415..432
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     416..432
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    448..547
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3815..3914)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3931..3950)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3932..3950)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3955..3971)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3955..3971)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4402..5193
                     /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      5286..5874
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5775..5794
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6028..6045
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
       61 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct
      121 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
      181 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca
      241 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
      301 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg
      361 cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa
      421 acgacggcca gtcttaagct cgggccccaa ataatgattt tattttgact gatagtgacc
      481 tgttcgttgc aacaaattga tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa
      541 gttggcacca tggggacttc ccatccggcg ttcctggtct taggctgtct tctcacaggg
      601 ctgagcctaa tcctctgcca gctttcatta ccctctatcc ttccaaatga aaatgaaaag
      661 gttgtgcagc tgaattcatc cttttctctg agatgctttg gggagagtga agtgagctgg
      721 cagtacccca tgtctgaaga agagagctcc gatgtggaaa tcagaaatga agaaaacaac
      781 agcggccttt ttgtgacggt cttggaagtg agcagtgcct cggcggccca cacagggttg
      841 tacacttgct attacaacca cactcagaca gaagagaatg agcttgaagg caggcacatt
      901 tacatctatg tgccagaccc agatgtagcc tttgtacctc taggaatgac ggattattta
      961 gtcatcgtgg aggatgatga ttctgccatt ataccttgtc gcacaactga tcccgagact
     1021 cctgtaacct tacacaacag tgagggggtg gtacctgcct cctacgacag cagacagggc
     1081 tttaatggga ccttcactgt agggccctat atctgtgagg ccaccgtcaa aggaaagaag
     1141 ttccagacca tcccatttaa tgtttatgct ttaaaagcaa catcagagct ggatctagaa
     1201 atggaagctc ttaaaaccgt gtataagtca ggggaaacga ttgtggtcac ctgtgctgtt
     1261 tttaacaatg aggtggttga ccttcaatgg acttaccctg gagaagtgaa aggcaaaggc
     1321 atcacaatac tggaagaaat caaagtccca tccatcaaat tggtgtacac tttgacggtc
     1381 cccgaggcca cggtgaaaga cagtggagat tacgaatgtg ctgcccgcca ggctaccagg
     1441 gaggtcaaag aaatgaagaa agtcactatt tctgtccatg agaaaggttt cattgaaatc
     1501 aaacccacct tcagccagtt ggaagctgtc aacctgcatg aagtcaaaca ttttgttgta
     1561 gaggtgcggg cctacccacc tcccaggata tcctggctga aaaacaatct gactctgatt
     1621 gaaaatctca ctgagatcac cactgatgtg gaaaagattc aggaaataag gtatcgaagc
     1681 aaattaaagc tgatccgtgc taaggaagaa gacagtggcc attatactat tgtagctcaa
     1741 aatgaagatg ctgtgaagag ctatactttt gaactgttaa ctcaagttcc ttcatccatt
     1801 ctggacttgg tcgatgatca ccatggctca actgggggac agacggtgag gtgcacagct
     1861 gaaggcacgc cgcttcctga tattgagtgg atgatatgca aagatattaa gaaatgtaat
     1921 aatgaaactt cctggactat tttggccaac aatgtctcaa acatcatcac ggagatccac
     1981 tcccgagaca ggagtaccgt ggagggccgt gtgactttcg ccaaagtgga ggagaccatc
     2041 gccgtgcgat gcctggctaa gaatctcctt ggagctgaga accgagagct gaagctggtg
     2101 gctcccaccc tgcgttctga actcacggtg gctgctgcag tcctggtgct gttggtgatt
     2161 gtgatcatct cacttattgt cctggttgtc atttggaaac agaaaccgag gtatgaaatt
     2221 cgctggaggg tcattgaatc aatcagcccg gatggacatg aatatattta tgtggacccg
     2281 atgcagctgc cttatgactc aagatgggag tttccaagag atggactagt gcttggtcgg
     2341 gtcttggggt ctggagcgtt tgggaaggtg gttgaaggaa cagcctatgg attaagccgg
     2401 tcccaacctg tcatgaaagt tgcagtgaag atgctaaaac ccacggccag atccagtgaa
     2461 aaacaagctc tcatgtctga actgaagata atgactcacc tggggccaca tttgaacatt
     2521 gtaaacttgc tgggagcctg caccaagtca ggccccattt acatcatcac agagtattgc
     2581 ttctatggag atttggtcaa ctatttgcat aagaataggg atagcttcct gagccaccac
     2641 ccagagaagc caaagaaaga gctggatatc tttggattga accctgctga tgaaagcaca
     2701 cggagctatg ttattttatc ttttgaaaac aatggtgact acatggacat gaagcaggct
     2761 gatactacac agtatgtccc catgctagaa aggaaagagg tttctaaata ttccgacatc
     2821 cagagatcac tctatgatcg tccagcctca tataagaaga aatctatgtt agactcagaa
     2881 gtcaaaaacc tcctttcaga tgataactca gaaggcctta ctttattgga tttgttgagc
     2941 ttcacctatc aagttgcccg aggaatggag tttttggctt caaaaaattg tgtccaccgt
     3001 gatctggctg ctcgcaacgt cctcctggca caaggaaaaa ttgtgaagat ctgtgacttt
     3061 ggcctggcca gagacatcat gcatgattcg aactatgtgt cgaaaggcag tacctttctg
     3121 cccgtgaagt ggatggctcc tgagagcatc tttgacaacc tctacaccac actgagtgat
     3181 gtctggtctt atggcattct gctctgggag atcttttccc ttggtggcac cccttacccc
     3241 ggcatgatgg tggattctac tttctacaat aagatcaaga gtgggtaccg gatggccaag
     3301 cctgaccacg ctaccagtga agtctacgag atcatggtga aatgctggaa cagtgagccg
     3361 gagaagagac cctcctttta ccacctgagt gagattgtgg agaatctgct gcctggacaa
     3421 tataaaaaga gttatgaaaa aattcacctg gacttcctga agagtgacca tcctgctgtg
     3481 gcacgcatgc gtgtggactc agacaatgca tacattggtg tcacctacaa aaacgaggaa
     3541 gacaagctga aggactggga gggtggtctg gatgagcaga gactgagcgc tgacagtggc
     3601 tacatcattc ctctgcctga cattgaccct gtccctgagg aggaggacct gggcaagagg
     3661 aacagacaca gctcgcagac ctctgaagag agtgccattg agacgggttc cagcagttcc
     3721 accttcatca agagagagga cgagaccatt gaagacatcg acatgatgga cgacatcggc
     3781 atagactctt cagacctggt ggaagacagc ttcctgccaa ctttcttgta caaagttggc
     3841 attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat
     3901 aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata
     3961 gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
     4021 aaaaatatat catcatgcct cctctagacc agccaggaca gaaatgcctc gacttcgctg
     4081 ctacccaagg ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg aacccagtgg
     4141 acataagcct gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg
     4201 tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt catggcttgt
     4261 tatgactgtt tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac
     4321 gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc
     4381 cctaaaacaa agttaaacat tatgagggaa gcggtgatcg ccgaagtatc gactcaacta
     4441 tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg
     4501 tacggctccg cagtggatgg cggcctgaag ccacacagtg atattgattt gctggttacg
     4561 gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact
     4621 tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac
     4681 gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag
     4741 cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc
     4801 ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc
     4861 tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg
     4921 aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt
     4981 tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg
     5041 gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga
     5101 caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg
     5161 aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga gccacccatg accaaaatcc
     5221 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5281 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5341 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5401 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5461 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5521 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5581 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5641 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5701 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5761 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5821 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5881 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5941 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     6001 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagag
//
