LOCUS       sequence-456581-        4305 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 4305)
  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 4305)
  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-HCK
FEATURES             Location/Qualifiers
     source          1..4305
                     /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"
     protein_bind    complement(2184..2283)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2300..2319)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2301..2319)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2324..2340)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2324..2340)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2771..3562
                     /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      3655..4243
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4144..4163
                     /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 agttggcacc atggggtgca tgaagtccaa gttcctccag gtcggaggca atacattctc
      721 aaaaactgaa accagcgcca gcccacactg tcctgtgtac gtgccggatc ccacatccac
      781 catcaagccg gggcctaata gccacaacag caacacacca ggaatcaggg aggcaggctc
      841 tgaggacatc atcgtggttg ccctgtatga ttacgaggcc attcaccacg aagacctcag
      901 cttccagaag ggggaccaga tggtggtcct agaggaatcc ggggagtggt ggaaggctcg
      961 atccctggcc acccggaagg agggctacat cccaagcaac tatgtcgccc gcgttgactc
     1021 tctggagaca gaggagtggt ttttcaaggg catcagccgg aaggacgcag agcgccaact
     1081 gctggctccc ggcaacatgc tgggctcctt catgatccgg gatagcgaga ccactaaagg
     1141 aagctactct ttgtccgtgc gagactacga ccctcggcag ggagataccg tgaaacatta
     1201 caagatccgg accctggaca acgggggctt ctacatatcc ccccgaagca ccttcagcac
     1261 tctgcaggag ctggtggacc actacaagaa ggggaacgac gggctctgcc agaaactgtc
     1321 ggtgccctgc atgtcttcca agccccagaa gccttgggag aaagatgcct gggagatccc
     1381 tcgggaatcc ctcaagctgg agaagaaact tggagctggg cagtttgggg aagtctggat
     1441 ggccacctac aacaagcaca ccaaggtggc agtgaagacg atgaagccag ggagcatgtc
     1501 ggtggaggcc ttcctggcag aggccaacgt gatgaaaact ctgcagcatg acaagctggt
     1561 caaacttcat gcggtggtca ccaaggagcc catctacatc atcacggagt tcatggccaa
     1621 aggaagcttg ctggactttc tgaaaagtga tgagggcagc aagcagccat tgccaaaact
     1681 cattgacttc tcagcccaga ttgcagaagg catggccttc atcgagcaga ggaactacat
     1741 ccaccgagac ctccgagctg ccaacatctt ggtctctgca tccctggtgt gtaagattgc
     1801 tgactttggc ctggcccggg tcattgagga caacgagtac acggctcggg aaggggccaa
     1861 gttccccatc aagtggacag ctcctgaagc catcaacttt ggctccttca ccatcaagtc
     1921 agacgtctgg tcctttggta tcctgctgat ggagatcgtc acctacggcc ggatccctta
     1981 cccagggatg tcaaaccctg aagtgatccg agctctggag cgtggatacc ggatgcctcg
     2041 cccagagaac tgcccagagg agctctacaa catcatgatg cgctgctgga aaaaccgtcc
     2101 ggaggagcgg ccgaccttcg aatacatcca gagtgtgctg gatgacttct acacggccac
     2161 agagagccag taccaacagc agccaccaac tttcttgtac aaagttggca ttataagaaa
     2221 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     2281 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     2341 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     2401 atcatgcctc ctctagacca gccaggacag aaatgcctcg acttcgctgc tacccaaggt
     2461 tgccgggtga cgcacaccgt ggaaacggat gaaggcacga acccagtgga cataagcctg
     2521 ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca cgcaactggt ccagaacctt
     2581 gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt atgactgttt
     2641 ttttggggta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg
     2701 atgtttgatg ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa
     2761 gttaaacatt atgagggaag cggtgatcgc cgaagtatcg actcaactat cagaggtagt
     2821 tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt acggctccgc
     2881 agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg tgaccgtaag
     2941 gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt cggcttcccc
     3001 tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg acgacatcat
     3061 tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc gcaatgacat
     3121 tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct tgctgacaaa
     3181 agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct ttgatccggt
     3241 tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga actcgccgcc
     3301 cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc
     3361 agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc
     3421 ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac aagaagaaga
     3481 tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga aaggcgagat
     3541 caccaaggta gtcggcaaat aaccctcgag ccacccatga ccaaaatccc ttaacgtgag
     3601 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3661 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3721 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3781 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3841 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3901 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3961 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4021 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4081 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4141 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4201 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4261 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtt
//
