LOCUS       sequence-261665-        4526 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 4526)
  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 4526)
  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-MAP3K7
FEATURES             Location/Qualifiers
     source          1..4526
                     /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"
     CDS             complement(1486..1497)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(2405..2504)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2521..2540)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2522..2540)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2545..2561)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2545..2561)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2992..3783
                     /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      3876..4464
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4365..4384
                     /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 agttggcatg tctacagcct ctgccgcctc ctcctcctcc tcgtcttcgg ccggtgagat
      721 gatcgaagcc ccttcccagg tcctcaactt tgaagagatc gactacaagg agatcgaggt
      781 ggaagaggtt gttggaagag gagcctttgg agttgtttgc aaagctaagt ggagagcaaa
      841 agatgttgct attaaacaaa tagaaagtga atctgagagg aaagcgttta ttgtagagct
      901 tcggcagtta tcccgtgtga accatcctaa tattgtaaag ctttatggag cctgcttgaa
      961 tccagtgtgt cttgtgatgg aatatgctga agggggctct ttatataatg tgctgcatgg
     1021 tgctgaacca ttgccatatt atactgctgc ccacgcaatg agttggtgtt tacagtgttc
     1081 ccaaggagtg gcttatcttc acagcatgca acccaaagcg ctaattcaca gggacctgaa
     1141 accaccaaac ttactgctgg ttgcaggggg gacagttcta aaaatttgtg attttggtac
     1201 agcctgtgac attcagacac acatgaccaa taacaagggg agtgctgctt ggatggcacc
     1261 tgaagttttt gaaggtagta attacagtga aaaatgtgac gtcttcagct ggggtattat
     1321 tctttgggaa gtgataacgc gtcggaaacc ctttgatgag attggtggcc cagctttccg
     1381 aatcatgtgg gctgttcata atggtactcg accaccactg ataaaaaatt tacctaagcc
     1441 cattgagagc ctgatgactc gttgttggtc taaagatcct tcccagcgcc cttcaatgga
     1501 ggaaattgtg aaaataatga ctcacttgat gcggtacttt ccaggagcag atgagccatt
     1561 acagtatcct tgtcagtatt cagatgaagg acagagcaac tctgccacca gtacaggctc
     1621 attcatggac attgcttcta caaatacgag taacaaaagt gacactaata tggagcaagt
     1681 tcctgccaca aatgatacta ttaagcgctt agaatcaaaa ttgttgaaaa atcaggcaaa
     1741 gcaacagagt gaatctggac gtttaagctt gggagcctcc cgtgggagca gtgtggagag
     1801 cttgccccca acctctgagg gcaagaggat gagtgctgac atgtctgaaa tagaagctag
     1861 gatcgccgca accacaggca acggacagcc aagacgtaga tccatccaag acttgactgt
     1921 aactggaaca gaacctggtc aggtgagcag taggtcatcc agtcccagtg tcagaatgat
     1981 tactacctca ggaccaacct cagaaaagcc aactcgaagt catccatgga cccctgatga
     2041 ttccacagat accaatggat cagataactc catcccaatg gcttatctta cactggatca
     2101 ccaactacag cctctagcac cgtgcccaaa ctccaaagaa tctatggcag tgtttgaaca
     2161 gcattgtaaa atggcacaag aatatatgaa agttcaaaca gaaattgcat tgttattaca
     2221 gagaaagcaa gaactagttg cagaactgga ccaggatgaa aaggaccagc aaaatacatc
     2281 tcgcctggta caggaacata aaaagctttt agatgaaaac aaaagccttt ctacttacta
     2341 ccagcaatgc aaaaaacaac tagaggtcat cagaagtcag cagcagaaac gacaaggcac
     2401 ttcatcccaa ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
     2461 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc
     2521 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct
     2581 caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgcct cctctagacc
     2641 agccaggaca gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg acgcacaccg
     2701 tggaaacgga tgaaggcacg aacccagtgg acataagcct gttcggttcg taagctgtaa
     2761 tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt
     2821 aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg
     2881 cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc
     2941 agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat tatgagggaa
     3001 gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat
     3061 ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag
     3121 ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg
     3181 cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc
     3241 cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct
     3301 aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag
     3361 ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt
     3421 gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt
     3481 gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag
     3541 cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg
     3601 ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc
     3661 atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca
     3721 gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa
     3781 taaccctcga gccacccatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     3841 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3901 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3961 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4021 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4081 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4141 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4201 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4261 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4321 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4381 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4441 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4501 ttttgctggc cttttgctca catgtt
//
