LOCUS       sequence-298117-        5193 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 5193)
  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 5193)
  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-PIK4CB
FEATURES             Location/Qualifiers
     source          1..5193
                     /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(3072..3171)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3188..3207)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3189..3207)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3212..3228)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3212..3228)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3659..4450
                     /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      4543..5131
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5032..5051
                     /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 agttggcatg ggagatacag tagtggagcc tgcccccttg aagccaactt ctgagcccac
      721 ttctggccca ccagggaata atggggggtc cctgctaagt gtcatcacgg agggggtcgg
      781 ggaactatca gtgattgacc ctgaggtggc ccagaaggcc tgccaggagg tgttggagaa
      841 agtcaagctt ttgcatggag gcgtggcagt ctctagcaga ggcaccccac tggagttggt
      901 caatggggat ggtgtggaca gtgagatccg ttgcctagat gatccacctg cccagatcag
      961 ggaggaggaa gatgagatgg gggccgctgt ggcctcaggc acagccaaag gagcaagaag
     1021 acggcggcag aacaactcag ctaaacagtc ttggctgctg aggctgtttg agtcaaaact
     1081 gtttgacatc tccatggcca tttcatacct gtataactcc aaggagcctg gagtacaagc
     1141 ctacattggc aaccggctct tctgctttcg caacgaggac gtggacttct atctgcccca
     1201 gttgcttaac atgtacatcc acatggatga ggacgtgggt gatgccatta agccctacat
     1261 agtccaccgt tgccgccaga gcattaactt ttccctccag tgtgccctgt tgcttggggc
     1321 ctattcttca gacatgcaca tttccactca acgacactcc cgtgggacca agctacggaa
     1381 gctgatcctc tcagatgagc taaagccagc tcacaggaag agggagctgc cctccttgag
     1441 cccggcccct gatacagggc tgtctccctc caaaaggact caccagcgct ctaagtcaga
     1501 tgccactgcc agcataagtc tcagcagcaa cctgaaacga acagccagca accctaaagt
     1561 ggagaatgag gatgagcctg ttcgactggc tcctgagaga gaattcatca agtccctgat
     1621 ggcgatcggc aagcggctgg ccacgctccc caccaaagag cagaaaacac agaggctgat
     1681 ctcagagctc tccctgctca accataagct ccctgcccga gtctggctgc ccactgctgg
     1741 ctttgaccac cacgtggtcc gtgtacccca cacacaggct gttgtcctca actccaagga
     1801 caaggctccc tacctgattt atgtggaagt ccttgaatgt gaaaactttg acaccaccag
     1861 tgtccctgcc cggatccccg agaaccgaat tcggagtacg aggtccgtag aaaacttgcc
     1921 cgaatgtggt attacccatg agcagcgagc tggcagcttc agcactgtgc ccaactatga
     1981 caacgatgat gaggcctggt cggtggatga cataggcgag ctgcaagtgg agctccccga
     2041 agtgcatacc aacagctgtg acaacatctc ccagttctct gtggacagca tcaccagcca
     2101 ggagagcaag gagcctgtgt tcattgcagc aggggacatc cgccggcgcc tttcggaaca
     2161 gctggctcat accccgacag ccttcaaacg agacccagaa gatccttctg cagttgctct
     2221 caaagagccc tggcaggaga aagtacggcg gatcagagag ggctccccct acggccatct
     2281 ccccaattgg cggctcctgt cagtcattgt caagtgtggg gatgaccttc ggcaagagct
     2341 tctggccttt caggtgttga agcaactgca gtccatttgg gaacaggagc gagtgcccct
     2401 ttggatcaag ccatacaaga ttcttgtgat ttcggctgat agtggcatga ttgaaccagt
     2461 ggtcaatgct gtgtccatcc atcaggtgaa gaaacagtca cagctctcct tgctcgatta
     2521 cttcctacag gagcacggca gttacaccac tgaggcattc ctcagtgcac agcgcaattt
     2581 tgtgcaaagt tgtgctgggt actgcttggt ctgctacctg ctgcaagtca aggacagaca
     2641 caatgggaat atccttttgg acgcagaagg ccacatcatc cacatcgact ttggcttcat
     2701 cctctccagc tcaccccgaa atctgggctt tgagacgtca gcctttaagc tgaccacaga
     2761 gtttgtggat gtgatgggcg gcctggatgg cgacatgttc aactactata agatgctgat
     2821 gctgcaaggg ctgattgccg ctcggaaaca catggacaag gtggtgcaga tcgtggagat
     2881 catgcagcaa ggttctcagc ttccttgctt ccatggctcc agcaccattc gaaacctcaa
     2941 agagaggttc cacatgagca tgactgagga gcagctgcag ctgctggtgg agcagatggt
     3001 ggatggcagt atgcggtcta tcaccaccaa actctatgac ggcttccagt acctcaccaa
     3061 cgacatcatg tgcccaactt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     3121 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     3181 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     3241 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgcctcct
     3301 ctagaccagc caggacagaa atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg
     3361 cacaccgtgg aaacggatga aggcacgaac ccagtggaca taagcctgtt cggttcgtaa
     3421 gctgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag
     3481 cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttggggtaca
     3541 gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt
     3601 atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacattat
     3661 gagggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga
     3721 gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg
     3781 cctgaagcca cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac
     3841 aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga
     3901 gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta
     3961 tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat
     4021 cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca
     4081 tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga
     4141 tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg
     4201 cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa
     4261 aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca
     4321 gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc
     4381 gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt
     4441 cggcaaataa ccctcgagcc acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     4501 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     4561 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     4621 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4681 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4741 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4801 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4861 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4921 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4981 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5041 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5101 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5161 cctggccttt tgctggcctt ttgctcacat gtt
//
