LOCUS       sequence-468766-        4419 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 4419)
  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 4419)
  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-CHEK2
FEATURES             Location/Qualifiers
     source          1..4419
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     10..32
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     24..41
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     25..41
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    57..156
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1786..1885)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1902..1921)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1903..1921)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1926..1942)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1926..1942)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2373..3164
                     /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      3257..3845
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3746..3765
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3999..4016
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4175..4202)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4294..4380)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggatgttttc ccagtcacga cgttgtaaaa cgacggccag tcttaagctc gggccccaaa
       61 taatgatttt attttgactg atagtgacct gttcgttgca acaaattgat gagcaatgct
      121 tttttataat gccaactttg tacaaaaaag ttggcatgtc tcgggagtcg gatgttgagg
      181 ctcagcagtc tcatggcagc agtgcctgtt cacagcccca tggcagcgtt acccagtccc
      241 aaggctcctc ctcacagtcc cagggcatat ccagctcctc taccagcacg atgccaaact
      301 ccagccagtc ctctcactcc agctctggga cactgagctc cttagagaca gtgtccactc
      361 aggaactcta ttctattcct gaggaccaag aacctgagga ccaagaacct gaggagccta
      421 cccctgcccc ctgggctcga ttatgggccc ttcaggatgg atttgccaat cttgaatgtg
      481 tgaatgacaa ctactggttt gggagggaca aaagctgtga atattgcttt gatgaaccac
      541 tgctgaaaag aacagataaa taccgaacat acagcaagaa acactttcgg attttcaggg
      601 aagtgggtcc taaaaactct tacattgcat acatagaaga tcacagtggc aatggaacct
      661 ttgtaaatac agagcttgta gggaaaggaa aacgccgtcc tttgaataac aattctgaaa
      721 ttgcactgtc actaagcaga aataaagttt ttgtcttttt tgatctgact gtagatgatc
      781 agtcagttta tcctaaggca ttaagagatg aatacatcat gtcaaaaact cttggaagtg
      841 gtgcctgtgg agaggtaaag ctggctttcg agaggaaaac atgtaagaaa gtagccataa
      901 agatcatcag caaaaggaag tttgctattg gttcagcaag agaggcagac ccagctctca
      961 atgttgaaac agaaatagaa attttgaaaa agctaaatca tccttgcatc atcaagatta
     1021 aaaacttttt tgatgcagaa gattattata ttgttttgga attgatggaa gggggagagc
     1081 tgtttgacaa agtggtgggg aataaacgcc tgaaagaagc tacctgcaag ctctattttt
     1141 accagatgct cttggctgtg cagtaccttc atgaaaacgg tattatacac cgtgacttaa
     1201 agccagagaa tgttttactg tcatctcaag aagaggactg tcttataaag attactgatt
     1261 ttgggcactc caagattttg ggagagacct ctctcatgag aaccttatgt ggaaccccca
     1321 cctacttggc gcctgaagtt cttgtttctg ttgggactgc tgggtataac cgtgctgtgg
     1381 actgctggag tttaggagtt attcttttta tctgccttag tgggtatcca cctttctctg
     1441 agcataggac tcaagtgtca ctgaaggatc agatcaccag tggaaaatac aacttcattc
     1501 ctgaagtctg ggcagaagtc tcagagaaag ctctggacct tgtcaagaag ttgttggtag
     1561 tggatccaaa ggcacgtttt acgacagaag aagccttaag acacccgtgg cttcaggatg
     1621 aagacatgaa gagaaagttt caagatcttc tgtctgagga aaatgaatcc acagctctac
     1681 cccaggttct agcccagcct tctactagtc gaaagcggcc ccgtgaaggg gaagccgagg
     1741 gtgccgagac cacaaagcgc ccagctgtgt gtgctgctgt gttgtgccca actttcttgt
     1801 acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta
     1861 tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt
     1921 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca
     1981 ttgcacaaga taaaaatata tcatcatgcc tcctctagac cagccaggac agaaatgcct
     2041 cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac
     2101 gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct
     2161 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt
     2221 tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc
     2281 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc
     2341 agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat
     2401 cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg
     2461 ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt
     2521 tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc
     2581 ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca
     2641 ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg
     2701 gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg
     2761 atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg
     2821 cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct
     2881 taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt
     2941 tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg
     3001 actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg
     3061 cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg
     3121 tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat
     3181 gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa
     3241 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3301 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3361 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     3421 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3481 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3541 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3601 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     3661 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3721 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3781 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3841 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3901 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     3961 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     4021 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     4081 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc
     4141 gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt
     4201 ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg
     4261 ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac
     4321 cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg
     4381 atgcctggca gttccctact ctcgcgttaa cgctagcat
//
