LOCUS       sequence-468767-        4089 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 4089)
  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 4089)
  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-STK11
FEATURES             Location/Qualifiers
     source          1..4089
                     /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(1456..1555)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1572..1591)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1573..1591)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1596..1612)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1596..1612)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2043..2834
                     /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      2927..3515
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3416..3435
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3669..3686
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3845..3872)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3964..4050)
                     /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 ttggcatgga ggtggtggac ccgcagcagc
      181 tgggcatgtt cacggagggc gagctgatgt cggtgggtat ggacacgttc atccaccgca
      241 tcgactccac cgaggtcatc taccagccgc gccgcaagcg ggccaagctc atcggcaagt
      301 acctgatggg ggacctgctg ggggaaggct cttacggcaa ggtgaaggag gtgctggact
      361 cggagacgct gtgcaggagg gccgtcaaga tcctcaagaa gaagaagttg cgaaggatcc
      421 ccaacgggga ggccaacgtg aagaaggaaa ttcaactact gaggaggtta cggcacaaaa
      481 atgtcatcca gctggtggat gtgttataca acgaagagaa gcagaaaatg tatatggtga
      541 tggagtactg cgtgtgtggc atgcaggaaa tgctggacag cgtgccggag aagcgtttcc
      601 cagtgtgcca ggcccacggg tacttctgtc agctgattga cggcctggag tacctgcata
      661 gccagggcat tgtgcacaag gacatcaagc cggggaacct gctgctcacc accggtggca
      721 ccctcaaaat ctccgacctg ggcgtggccg aggcactgca cccgttcgcg gcggacgaca
      781 cctgccggac cagccagggc tccccggctt tccagccgcc cgagattgcc aacggcctgg
      841 acaccttctc cggcttcaag gtggacatct ggtcggctgg ggtcaccctc tacaacatca
      901 ccacgggtct gtaccccttc gaaggggaca acatctacaa gttgtttgag aacatcggga
      961 aggggagcta cgccatcccg ggcgactgtg gccccccgct ctctgacctg ctgaaaggga
     1021 tgcttgagta cgaaccggcc aagaggttct ccatccggca gatccggcag cacagctggt
     1081 tccggaagaa acatcctccg gctgaagcac cagtgcccat cccaccgagc ccagacacca
     1141 aggaccggtg gcgcagcatg actgtggtgc cgtacttgga ggacctgcac ggcgcggacg
     1201 aggacgagga cctcttcgac atcgaggatg acatcatcta cactcaggac ttcacggtgc
     1261 ccggacaggt cccagaagag gaggccagtc acaatggaca gcgccggggc ctccccaagg
     1321 ccgtgtgtat gaacggcaca gaggcggcgc agctgagcac caaatccagg gcggagggcc
     1381 gggcccccaa ccctgcccgc aaggcctgct ccgccagcag caagatccgc cggctgtcgg
     1441 cctgcaagca gcagtgccca actttcttgt acaaagttgg cattataaga aagcattgct
     1501 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc
     1561 cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
     1621 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgcc
     1681 tcctctagac cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt
     1741 gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc
     1801 gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
     1861 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg
     1921 tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga
     1981 tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca
     2041 ttatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca
     2101 tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg
     2161 gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg
     2221 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga
     2281 gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc
     2341 gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag
     2401 gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag
     2461 aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac
     2521 aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg
     2581 ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg
     2641 gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt
     2701 atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg
     2761 cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg
     2821 tagtcggcaa ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt
     2881 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2941 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3001 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3061 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3121 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3181 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3241 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3301 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3361 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3421 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3481 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3541 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     3601 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     3661 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     3721 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     3781 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
     3841 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
     3901 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
     3961 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     4021 gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa
     4081 cgctagcat
//
