LOCUS       sequence-375327-        3153 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 3153)
  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 3153)
  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-PNCK
FEATURES             Location/Qualifiers
     source          1..3153
                     /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(464..563)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(580..599)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(581..599)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(604..620)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(604..620)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1051..1842
                     /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      1935..2523
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2424..2443
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2677..2694
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2853..2880)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2972..3058)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3107..3129
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3121..3138
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3122..3138
                     /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 tgctgctgct gaagaaacac
      121 acggaggaca tcagcagcgt ctacgagatc cgcgagaggc tcggctcggg gccctccccg
      181 ctccactctc tctccctcct gcctctcctc tcttcccact tccttcccac ctcccaccgc
      241 cccgtctgcg gcaggggtgc cttctccgag gtggtgctgg cccaggagcg gggctccgca
      301 cacctcgtgg ccctcaagtg catccccaag aaggccctcc ggggcaagga ggccctggtg
      361 gagaacgaga tcgcagtgct ccgtaggatc agtcacccca acatcgtcgc tctggaggat
      421 gtccacgaga gcccttccca cctctacctg gccatggaac tgtgcccaac tttcttgtac
      481 aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc
      541 agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac
      601 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt
      661 gcacaagata aaaatatatc atcatgcctc ctctagacca gccaggacag aaatgcctcg
      721 acttcgctgc tacccaaggt tgccgggtga cgcacaccgt ggaaacggat gaaggcacga
      781 acccagtgga cataagcctg ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca
      841 cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc
      901 atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa
      961 gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag
     1021 ggcagtcgcc ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc cgaagtatcg
     1081 actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc
     1141 gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg
     1201 ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt
     1261 ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt
     1321 gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga
     1381 gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat
     1441 ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg
     1501 gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta
     1561 acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg
     1621 tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac
     1681 tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct
     1741 tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc
     1801 cactacgtga aaggcgagat caccaaggta gtcggcaaat aaccctcgag ccacccatga
     1861 ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag
     1921 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     1981 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2041 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     2101 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2161 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2221 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2281 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2341 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2401 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2461 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2521 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2581 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     2641 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     2701 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     2761 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt
     2821 accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct
     2881 gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt
     2941 gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg
     3001 acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat
     3061 gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt
     3121 tgtaaaacga cggccagtct taagctcggg ccc
//
