LOCUS       sequence-459880-        5328 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 5328)
  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 5328)
  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-MAP4K5
FEATURES             Location/Qualifiers
     source          1..5328
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2639..2738)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2755..2774)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2756..2774)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2779..2795)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2779..2795)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3226..4017
                     /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      4110..4698
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4599..4618
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4852..4869
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5028..5055)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5147..5233)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5282..5304
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5296..5313
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5297..5313
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagttggca tggaggcccc gctgcggcct
      121 gccgcggaca tcctgaggcg gaacccgcag caggactacg aactcgtcca gagggtcggc
      181 agcggcacct acggggacgt ctataaggcc agaaatgtac acacaggaga gctggctgca
      241 gtaaaaatca ttaaattgga gcctggagat gatttttctt tgattcaaca agaaatattt
      301 atggttaaag aatgtaaaca ttgtaacatc gttgcctact ttgggagtta tcttagtcgg
      361 gaaaaactat ggatttgtat ggaatactgt ggtggcggat cacttcaaga tatttaccat
      421 gttactggac cattatcaga attgcaaata gcctatgtat gcagagaaac cttacagggt
      481 cttgcctatt tgcatactaa aggcaaaatg catagagata tcaaaggtgc taatatttta
      541 ttgacagacc atggcgatgt aaaattagct gactttggtg tggctgcaaa aataacagct
      601 accattgcaa aacgaaaatc tttcattggc accccttact ggatggcccc agaagttgca
      661 gcagtagaga agaatggtgg ctacaaccaa ctctgtgata tctgggcagt aggaataaca
      721 gcaattgaac ttggagaact tcagccacct atgtttgatc tccacccaat gagggctctc
      781 ttcttaatgt caaaaagtaa ttttcagcct ccaaaactaa aggacaaaac aaaatggtca
      841 tcaacattcc ataattttgt caaaatagca ctaaccaaaa acccaaaaaa aagaccaact
      901 gctgaaagac ttctgactca cacttttgtt gcacagccag gtctctctag agccctagca
      961 gttgaactgt tagacaaagt gaacaatcca gataaccacg cacattacac tgaagcagat
     1021 gacgatgact ttgagcccca tgcaatcatt cgtcatacca ttagatctac aaacaggaat
     1081 gccagagctg aacggacagc ttcagaaata aattttgaca aattacaatt tgaacctcct
     1141 ctgagaaaag aaacagaagc acgagatgaa atgggattgt catcagaccc aaatttcatg
     1201 ttacagtgga atccttttgt tgatggtgca aatactggca aatcaacctc aaaacgtgca
     1261 ataccacctc ccctacctcc taagccaagg ataagcagtt accctgaaga caactttccg
     1321 gatgaagaaa aagcatcaac cataaaacat tgtcctgatt cagaaagcag agctccccaa
     1381 attctcagaa gacagagtag cccaagttgt gggcctgtgg cagagacttc ttctattgga
     1441 aatggtgatg gtatttcaaa actgatgagt gaaaatacag aaggatcagc acaagcacca
     1501 cagttaccac gaaaaaagga caaacgagac ttccctaaac cagccatcaa tggccttcca
     1561 cccaccccaa aagttctgat gggagcatgc ttttcaaaag tttttgatgg ctgtcctttg
     1621 aaaattaatt gtgcaacatc ctggatacat cctgatacaa aagatcagta cattattttt
     1681 ggaactgaag atggtattta cacactgaat ctcaatgagc tacatgaggc aacgatggaa
     1741 cagttatttc cacggaagtg tacttggctg tatgttatca ataatacttt aatgtcatta
     1801 tcagaaggaa aaacctttca gctctactct cacaatctta tagctttgtt tgaacatgcc
     1861 aaaaaaccag gattagctgc ccatattcaa actcacaggt ttccagaccg aatactacca
     1921 agaaaattcg ctttaacaac aaagattcct gatacaaaag gctgccacaa atgttgcata
     1981 gtcagaaacc cttacacggg acataaatac ctctgtggag ctttacagtc tggaattgtt
     2041 ttacttcagt ggtatgagcc aatgcagaaa ttcatgttga taaagcactt tgattttcct
     2101 ttgccaagtc ctttgaatgt ttttgaaatg ctggtgatac ctgaacagga ataccctatg
     2161 gtctgtgtag ctattagcaa aggcactgaa tcgaatcagg tagttcagtt tgagacaatc
     2221 aatttgaact ctgcatcttc atggtttaca gaaattggtg caggcagcca gcagttagat
     2281 tccattcatg taacacagtt ggagagagat accgttttag tgtgtttaga caaatttgtg
     2341 aaaattgtaa atctacaagg aaaattaaaa tcaagtaaga aactggcctc tgagttaagt
     2401 tttgattttc gcattgaatc tgtagtatgc cttcaagaca gtgtgttggc tttctggaaa
     2461 catgggatgc agggtaaaag cttcaagtca gatgaggtta cccaggagat ttcagatgaa
     2521 acaagagttt tccgcttatt aggatcagac agggttgtcg ttttggaaag taggccaaca
     2581 gaaaatccta ctgcacacag caatctctac atcttggctg gacatgaaaa tagttactac
     2641 ccaactttct tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac
     2701 gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga tatcccctat
     2761 agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat
     2821 ctctgatgtt acattgcaca agataaaaat atatcatcat gcctcctcta gaccagccag
     2881 gacagaaatg cctcgacttc gctgctaccc aaggttgccg ggtgacgcac accgtggaaa
     2941 cggatgaagg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag
     3001 tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc
     3061 gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc tatgcctcgg
     3121 gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac
     3181 gatgttacgc agcagggcag tcgccctaaa acaaagttaa acattatgag ggaagcggtg
     3241 atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa
     3301 ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac
     3361 agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct
     3421 ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct
     3481 gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc
     3541 gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc
     3601 acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg
     3661 gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg
     3721 ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat
     3781 gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag
     3841 gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt
     3901 gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag
     3961 ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataaccc
     4021 tcgagccacc catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca
     4081 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4141 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4201 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4261 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4321 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4381 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4441 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4501 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4561 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4621 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4681 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4741 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4801 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     4861 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     4921 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     4981 gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg
     5041 tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc
     5101 accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca
     5161 ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct
     5221 ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt
     5281 tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggccc
//
