LOCUS       sequence-137987-        4008 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 4008)
  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 4008)
  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-PIP5K2A
FEATURES             Location/Qualifiers
     source          1..4008
                     /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(1768..1867)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1884..1903)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1885..1903)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1908..1924)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1908..1924)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2355..3146
                     /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      3239..3827
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3728..3747
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3981..3998
                     /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 aagttggcat ggcgaccccc ggcaacctag ggtcctctgt cctggcgagc aagaccaaga
      601 ccaagaagaa gcacttcgta gcgcagaaag tgaagctgtt tcgggccagc gacccgctgc
      661 tcagcgtcct catgtggggg gtaaaccact cgatcaatga actgagccat gttcaaatcc
      721 ctgttatgtt gatgccagat gacttcaaag cctattcaaa aataaaggtg gacaatcacc
      781 tttttaacaa agaaaacatg ccgagccatt tcaagtttaa ggaatactgc ccgatggtct
      841 tccgtaacct gcgggagagg tttggaattg atgatcaaga tttccagaat tccctgacca
      901 ggagcgcacc cctccccaac gactcccagg cccgcagtgg agctcgtttt cacacttcct
      961 acgacaaaag atacatcatc aagactatta ccagtgaaga cgtggccgaa atgcacaaca
     1021 tcctgaagaa ataccaccag tacatagtgg aatgtcatgg gatcaccctt cttccccagt
     1081 tcttgggcat gtaccggctt aatgttgatg gagttgaaat atatgtgata gttacaagaa
     1141 atgtattcag ccaccgtttg tctgtgtata ggaaatacga cttaaagggc tctacagtgg
     1201 ctagagaagc tagtgacaaa gaaaaggcca aagaactgcc aactctgaaa gataatgatt
     1261 tcattaatga gggccaaaag atttatattg atgacaacaa caagaaggtc ttcctggaaa
     1321 aactaaaaaa ggatgttgag tttctggccc agctgaagct catggactac agtctgctgg
     1381 tgggaattca tgatgtggag agagccgaac aggaggaagt ggagtgtgag gagaacgatg
     1441 gggaggagga gggcgagagc gatggcaccc acccggtggg aaccccccca gatagccccg
     1501 ggaatacact gaacagctca ccacccctgg ctcccgggga gttcgatccg aacatcgacg
     1561 tctatggaat taagtgccat gaaaactcgc ctaggaagga ggtgtacttc atggcaatta
     1621 ttgacatcct tactcattat gatgcaaaaa agaaagctgc ccatgctgca aaaactgtta
     1681 aacatggcgc tggcgcggag atctccaccg tgaacccaga acagtattca aagcgctttt
     1741 tggactttat tggccacatc ttgacgtacc caactttctt gtacaaagtt ggcattataa
     1801 gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca
     1861 ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt
     1921 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     1981 tatcatcatg cctcctctag accagccagg acagaaatgc ctcgacttcg ctgctaccca
     2041 aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag tggacataag
     2101 cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa
     2161 ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact
     2221 gtttttttgg ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg
     2281 gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa
     2341 caaagttaaa cattatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg
     2401 tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct
     2461 ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg
     2521 taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt
     2581 cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca
     2641 tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg
     2701 acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga
     2761 caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc
     2821 cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc
     2881 cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca
     2941 gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc
     3001 tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag
     3061 aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg
     3121 agatcaccaa ggtagtcggc aaataaccct cgagccaccc atgaccaaaa tcccttaacg
     3181 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3241 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3301 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3361 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3421 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3481 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3541 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3601 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3661 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3721 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3781 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3841 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3901 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3961 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
