LOCUS       sequence-263695-        3933 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 3933)
  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 3933)
  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-CKB
FEATURES             Location/Qualifiers
     source          1..3933
                     /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(1244..1343)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1360..1379)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1361..1379)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1384..1400)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1384..1400)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1831..2622
                     /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      2715..3303
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3204..3223
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3457..3474
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3633..3660)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3752..3838)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3887..3909
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3901..3918
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3902..3918
                     /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 tgcccttctc caacagccac
      121 aacgcactga agctgcgctt cccggccgag gacgagttcc ccgacctgag cgcccacaac
      181 aaccacatgg ccaaggtgct gacccccgag ctgtacgcgg agctgcgcgc caagagcacg
      241 ccgagcggct tcacgctgga cgacgtcatc cagacaggcg tggacaaccc gggccacccg
      301 tacatcatga ccgtgggctg cgtggcgggc gacgaggagt cctacgaagt gttcaaggat
      361 ctcttcgacc ccatcatcga ggaccggcac ggcggctaca agcccagcga tgagcacaag
      421 accgacctca accccgacaa cctgcagggc ggcgacgacc tggaccccaa ctacgtgctg
      481 agctcgcggg tgcgcacggg ccgcagcatc cgtggcttct gcctcccccc gcactgcagc
      541 cgcggggagc gccgagccat cgagaagctc gcggtggaag ccctgtccag cctggacggc
      601 gacctggcgg gccgatacta cgcgctcaag agcatgacgg aggcggagca gcagcagctc
      661 atcgacgacc acttcctctt cgacaagccc gtgtcgcccc tgctgctggc ctcgggcatg
      721 gcccgcgact ggcccgacgc ccgcggtatc tggcacaatg acaataagac cttcctggtg
      781 tgggtcaacg aggaggacca cctgcgggtc atctccatgc agaagggggg caacatgaag
      841 gaggtgttca cccgcttctg caccggcctc acccagattg aaactctctt caagtctaag
      901 gactatgagt tcatgtggaa ccctcacctg ggctacatcc tcacctgccc atccaacctg
      961 ggcaccgggc tgcgggcagg tgtgcatatc aagctgccca acctgggcaa gcatgagaag
     1021 ttctcggagg tgcttaagcg gctgcgactt cagaagcgag gcacaggcgg tgtggacacg
     1081 gctgcggtgg gcggggtctt cgacgtctcc aacgctgacc gcctgggctt ctcagaggtg
     1141 gagctggtgc agatggtggt ggacggagtg aagctgctca tcgagatgga acagcggctg
     1201 gagcagggcc aggccatcga cgacctcatg cctgcccaga aatgcccaac tttcttgtac
     1261 aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc
     1321 agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac
     1381 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     1441 gcacaagata aaaatatatc atcatgcctc ctctagacca gccaggacag aaatgcctcg
     1501 acttcgctgc tacccaaggt tgccgggtga cgcacaccgt ggaaacggat gaaggcacga
     1561 acccagtgga cataagcctg ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca
     1621 cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc
     1681 atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa
     1741 gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag
     1801 ggcagtcgcc ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc cgaagtatcg
     1861 actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc
     1921 gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg
     1981 ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt
     2041 ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt
     2101 gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga
     2161 gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat
     2221 ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg
     2281 gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta
     2341 acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg
     2401 tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac
     2461 tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct
     2521 tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc
     2581 cactacgtga aaggcgagat caccaaggta gtcggcaaat aaccctcgag ccacccatga
     2641 ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag
     2701 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2761 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2821 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     2881 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2941 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3001 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3061 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3121 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3181 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3241 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3301 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3361 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3421 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3481 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     3541 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt
     3601 accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct
     3661 gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt
     3721 gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg
     3781 acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat
     3841 gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt
     3901 tgtaaaacga cggccagtct taagctcggg ccc
//
