LOCUS       sequence-300525-        5769 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 5769)
  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 5769)
  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-FLT3
FEATURES             Location/Qualifiers
     source          1..5769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3648..3747)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3764..3783)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3765..3783)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3788..3804)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3788..3804)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4235..5026
                     /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      5119..5707
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5608..5627
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agttggcacc atgccggcgt tggcgcgcga cggcggccag ctgccgctgc tcgttgtttt
      721 ttctgcaatg atatttggga ctattacaaa tcaagatctg cctgtgatca agtgtgtttt
      781 aatcaatcat aagaacaatg attcatcagt ggggaagtca tcatcatatc ccatggtatc
      841 agaatccccg gaagacctcg ggtgtgcgtt gagaccccag agctcaggga cagtgtacga
      901 agctgccgct gtggaagtgg atgtatctgc ttccatcaca ctgcaagtgc tggtcgatgc
      961 cccagggaac atttcctgtc tctgggtctt taagcacagc tccctgaatt gccagccaca
     1021 ttttgattta caaaacagag gagttgtttc catggtcatt ttgaaaatga cagaaaccca
     1081 agctggagaa tacctacttt ttattcagag tgaagctacc aattacacaa tattgtttac
     1141 agtgagtata agaaataccc tgctttacac attaagaaga ccttacttta gaaaaatgga
     1201 aaaccaggac gccctggtct gcatatctga gagcgttcca gagccgatcg tggaatgggt
     1261 gctttgcgat tcacaggggg aaagctgtaa agaagaaagt ccagctgttg ttaaaaagga
     1321 ggaaaaagtg cttcatgaat tatttgggat ggacataagg tgctgtgcca gaaatgaact
     1381 gggcagggaa tgcaccaggc tgttcacaat agatctaaat caaactcctc agaccacatt
     1441 gccacaatta tttcttaaag taggggaacc cttatggata aggtgcaaag ctgttcatgt
     1501 gaaccatgga ttcgggctca cctgggaatt agaaaacaaa gcactcgagg agggcaacta
     1561 ctttgagatg agtacctatt caacaaacag aactatgata cggattctgt ttgcttttgt
     1621 atcatcagtg gcaagaaacg acaccggata ctacacttgt tcctcttcaa agcatcccag
     1681 tcaatcagct ttggttacca tcgtagaaaa gggatttata aatgctacca attcaagtga
     1741 agattatgaa attgaccaat atgaagagtt ttgtttttct gtcaggttta aagcctaccc
     1801 acaaatcaga tgtacgtgga ccttctctcg aaaatcattt ccttgtgagc aaaagggtct
     1861 tgataacgga tacagcatat ccaagttttg caatcataag caccagccag gagaatatat
     1921 attccatgca gaaaatgatg atgcccaatt taccaaaatg ttcacgctga atataagaag
     1981 gaaacctcaa gtgctcgcag aagcatcggc aagtcaggcg tcctgtttct cggatggata
     2041 cccattacca tcttggacct ggaagaagtg ttcagacaag tctcccaact gcacagaaga
     2101 gatcacagaa ggagtctgga atagaaaggc taacagaaaa gtgtttggac agtgggtgtc
     2161 gagcagtact ctaaacatga gtgaagccat aaaagggttc ctggtcaagt gctgtgcgta
     2221 caattccctt ggcacatctt gtgagacgat ccttttaaac tctccaggcc ccttcccttt
     2281 catccaagac aacatctcat tctatgcaac aattggtgtt tgtctcctct tcattgtcgt
     2341 tttaaccctg ctaatttgtc acaagtacaa aaagcaattt aggtatgaaa gccagctaca
     2401 gatggtacag gtgaccggct cctcagataa tgagtacttc tacgttgatt tcagagaata
     2461 tgaatatgat ctcaaatggg agtttccaag agaaaattta gagtttggga aggtactagg
     2521 atcaggtgct tttggaaaag tgatgaacgc aacagcttat ggaattagca aaacaggagt
     2581 ctcaatccag gttgccgtca aaatgctgaa agaaaaagca gacagctctg aaagagaggc
     2641 actcatgtca gaactcaaga tgatgaccca gctgggaagc cacgagaata ttgtgaacct
     2701 gctgggggcg tgcacactgt caggaccaat ttacttgatt tttgaatact gttgctatgg
     2761 tgatcttctc aactatctaa gaagtaaaag agaaaaattt cacaggactt ggacagagat
     2821 tttcaaggaa cacaatttca gtttttaccc cactttccaa tcacatccaa attccagcat
     2881 gcctggttca agagaagttc agatacaccc ggactcggat caaatctcag ggcttcatgg
     2941 gaattcattt cactctgaag atgaaattga atatgaaaac caaaaaaggc tggaagaaga
     3001 ggaggacttg aatgtgctta catttgaaga tcttctttgc tttgcatatc aagttgccaa
     3061 aggaatggaa tttctggaat ttaagtcgtg tgttcacaga gacctggccg ccaggaacgt
     3121 gcttgtcacc cacgggaaag tggtgaagat atgtgacttt ggattggctc gagatatcat
     3181 gagtgattcc aactatgttg tcaggggcaa tgcccgtctg cctgtaaaat ggatggcccc
     3241 cgaaagcctg tttgaaggca tctacaccat taagagtgat gtctggtcat atggaatatt
     3301 actgtgggaa atcttctcac ttggtgtgaa tccttaccct ggcattccgg ttgatgctaa
     3361 cttctacaaa ctgattcaaa atggatttaa aatggatcag ccattttatg ctacagaaga
     3421 aatatacatt ataatgcaat cctgctgggc ttttgactca aggaaacggc catccttccc
     3481 taatttgact tcgtttttag gatgtcagct ggcagatgca gaagaagcga tgtatcagaa
     3541 tgtggatggc cgtgtttcgg aatgtcctca cacctaccaa aacaggcgac ctttcagcag
     3601 agagatggat ttggggctac tctctccgca ggctcaggtc gaagattcgc caactttctt
     3661 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     3721 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     3781 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     3841 cattgcacaa gataaaaata tatcatcatg cctcctctag accagccagg acagaaatgc
     3901 ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc
     3961 acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg
     4021 ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt
     4081 tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca
     4141 gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca
     4201 gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga tcgccgaagt
     4261 atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct
     4321 ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga
     4381 tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga
     4441 ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac
     4501 cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt
     4561 tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat
     4621 tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc
     4681 ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac
     4741 cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac
     4801 gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc
     4861 cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca
     4921 ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt
     4981 tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct cgagccaccc
     5041 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     5101 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5161 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5221 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5281 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5341 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5401 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5461 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5521 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5581 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5641 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5701 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5761 tcacatgtt
//
