LOCUS       sequence-359682-        4098 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 4098)
  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 4098)
  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-PDK1
FEATURES             Location/Qualifiers
     source          1..4098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(562..589)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(681..767)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     816..838
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     830..847
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     831..847
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    863..962
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2271..2370)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2387..2406)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2388..2406)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2411..2427)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2411..2427)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2858..3649
                     /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      join(3742..4098,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca
      541 ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga
      601 tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg
      661 ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag
      721 ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt
      781 ccctactctc gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac
      841 ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag tgacctgttc
      901 gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca aaaaagttgg
      961 catgaggctg gcgcggctgc ttcgcggagc cgccttggcc ggcccgggcc cggggctgcg
     1021 cgccgccggc ttcagccgca gcttcagctc ggactcgggc tccagcccgg cgtccgagcg
     1081 cggcgttccg ggccaggtgg acttctacgc gcgcttctcg ccgtccccgc tctccatgaa
     1141 gcagttcctg gacttcggat cagtgaatgc ttgtgaaaag acctcattta tgtttctgcg
     1201 gcaagagttg cctgtcagac tggcaaatat aatgaaagaa ataagtctcc ttccagataa
     1261 tcttctcagg acaccatccg ttcaattggt acaaagctgg tatatccaga gtcttcagga
     1321 gcttcttgat tttaaggaca aaagtgctga ggatgctaaa gctatttatg actttacaga
     1381 tactgtgata cggatcagaa accgacacaa tgatgtcatt cccacaatgg cccagggtgt
     1441 gattgaatac aaggagagct ttggggtgga tcctgtcacc agccagaatg ttcagtactt
     1501 tttggatcga ttctacatga gtcgcatttc aattagaatg ttactcaatc agcactcttt
     1561 attgtttggt ggaaaaggca aaggaagtcc atctcatcga aaacacattg gaagcataaa
     1621 tccaaactgc aatgtacttg aagttattaa agatggctat gaaaatgcta ggcgtctgtg
     1681 tgatttgtat tatattaact ctcccgaact agaacttgaa gaactaaatg caaaatcacc
     1741 aggacagcca atacaagtgg tttatgtacc atcccatctc tatcacatgg tgtttgaact
     1801 tttcaagaat gcaatgagag ccactatgga acaccatgcc aacagaggtg tttacccccc
     1861 tattcaagtt catgtcacgc tgggtaatga ggatttgact gtgaagatga gtgaccgagg
     1921 aggtggcgtt cctttgagga aaattgacag acttttcaac tacatgtatt caactgcacc
     1981 aagacctcgt gttgagacct cccgcgcagt gcctctggct ggttttggtt atggattgcc
     2041 catatcacgt ctttacgcac aatacttcca aggagacctg aagctgtatt ccctagaggg
     2101 ttacgggaca gatgcagtta tctacattaa ggctctgtca acagactcaa tagaaagact
     2161 cccagtgtat aacaaagctg cctggaagca ttacaacacc aaccacgagg ctgatgactg
     2221 gtgcgtcccc agcagagaac ccaaagacat gacgacgttc cgcagtgcct acccaacttt
     2281 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
     2341 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     2401 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     2461 ttacattgca caagataaaa atatatcatc atgcctcctc tagaccagcc aggacagaaa
     2521 tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc acaccgtgga aacggatgaa
     2581 ggcacgaacc cagtggacat aagcctgttc ggttcgtaag ctgtaatgca agtagcgtat
     2641 gcgctcacgc aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc
     2701 ggttttcatg gcttgttatg actgtttttt tggggtacag tctatgcctc gggcatccaa
     2761 gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta tggagcagca acgatgttac
     2821 gcagcagggc agtcgcccta aaacaaagtt aaacattatg agggaagcgg tgatcgccga
     2881 agtatcgact caactatcag aggtagttgg cgtcatcgag cgccatctcg aaccgacgtt
     2941 gctggccgta catttgtacg gctccgcagt ggatggcggc ctgaagccac acagtgatat
     3001 tgatttgctg gttacggtga ccgtaaggct tgatgaaaca acgcggcgag ctttgatcaa
     3061 cgaccttttg gaaacttcgg cttcccctgg agagagcgag attctccgcg ctgtagaagt
     3121 caccattgtt gtgcacgacg acatcattcc gtggcgttat ccagctaagc gcgaactgca
     3181 atttggagaa tggcagcgca atgacattct tgcaggtatc ttcgagccag ccacgatcga
     3241 cattgatctg gctatcttgc tgacaaaagc aagagaacat agcgttgcct tggtaggtcc
     3301 agcggcggag gaactctttg atccggttcc tgaacaggat ctatttgagg cgctaaatga
     3361 aaccttaacg ctatggaact cgccgcccga ctgggctggc gatgagcgaa atgtagtgct
     3421 tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa atcgcgccga aggatgtcgc
     3481 tgccgactgg gcaatggagc gcctgccggc ccagtatcag cccgtcatac ttgaagctag
     3541 acaggcttat cttggacaag aagaagatcg cttggcctcg cgcgcagatc agttggaaga
     3601 atttgtccac tacgtgaaag gcgagatcac caaggtagtc ggcaaataac cctcgagcca
     3661 cccatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     3721 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3781 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3841 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3901 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3961 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4021 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4081 gcccagcttg gagcgaac
//
