LOCUS       sequence-343771-        5271 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 5271)
  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 5271)
  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-PFKL
FEATURES             Location/Qualifiers
     source          1..5271
                     /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"
     regulatory      1879..1888
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2582..2681)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2698..2717)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2699..2717)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2722..2738)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2722..2738)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3169..3960
                     /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      4053..4641
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4542..4561
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4795..4812
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4971..4998)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5090..5176)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5225..5247
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5239..5256
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5240..5256
                     /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 tgtgtaacca gggtagaggt
      121 cgagagtcct ctcgtggggg tctccatgtt caagggagct gccgaggctt gagcaggagc
      181 ccccagcagg aaactggctt tgccaaggcc cccgctggga cagactgttt ctttcactgc
      241 agtcctggga gccgagggca aggggacagg aaagaggaag tgacctcaga gcctggtggc
      301 accagcatca tgtccaggct ggggggcatg aacgctgctg tccgggctgt gacgcgcatg
      361 ggcatttatg tgggtgccaa agtcttcctc atctacgagg gctatgaggg cctcgtggag
      421 ggaggtgaga acatcaagca ggccaactgg ctgagcgtct ccaacatcat ccagctgggc
      481 ggcactatca ttggcagcgc tcgctgcaag gcctttacca ccagggaggg gcgccgggca
      541 gcggcctaca acctggtcca gcacggcatc accaacctgt gcgtcatcgg cggggatggc
      601 agccttacag gtgccaacat cttccgcagc gagtggggca gcctgctgga ggagctggtg
      661 gcggaaggta agatctcaga gactacagcc cggacctact cgcacctgaa catcgcgggc
      721 ctagtgggct ccatcgataa cgacttctgc ggcaccgaca tgaccatcgg cacggactcg
      781 gccctccacc gcatcatgga ggtcatcgat gccatcacca ccactgccca gagccaccag
      841 aggaccttcg tgctggaagt gatgggccgg cactgcgggt acctggcgct ggtatctgca
      901 ctggcctcag gggccgactg gctgttcatc cccgaggctc cacccgagga cggctgggag
      961 aacttcatgt gtgagaggct gggtgagact cggagccgtg ggtcccgact gaacatcatc
     1021 atcatcgctg agggtgccat tgaccgcaac gggaagccca tctcgtccag ctacgtgaag
     1081 gacctggtgg ttcagaggct gggcttcgac acccgtgtaa ctgtgctggg ccacgtgcag
     1141 cggggaggga cgccctctgc cttcgaccgg atcctgagca gcaagatggg catggaggcg
     1201 gtgatggcgc tgctggaagc cacgcctgac acgccggcct gcgtggtcac cctctcgggg
     1261 aaccagtcag tgcggctgcc cctcatggag tgcgtgcaga tgaccaagga agtgcagaaa
     1321 gccatggatg acaagaggtt tgacgaggcc acccagctcc gtggtgggag cttcgagaac
     1381 aactggaaca tttacaagct cctcgcccac cagaagcccc ccaaggagaa gtctaacttc
     1441 tccctggcca tcctgaatgt gggggccccg gcggctggca tgaatgcggc cgtgcgctcg
     1501 gcggtgcgga ccggcatctc ccatggacac acagtatacg tggtgcacga tggcttcgaa
     1561 ggcctagcca agggtcaggt gcaagaagta ggctggcacg acgtggccgg ctggttgggg
     1621 cgtggtggct ccatgctggg gaccaagagg accctgccca agggccagct ggagtccatt
     1681 gtggagaaca tccgcatcta tggtattcac gccctgctgg tggtcggtgg gtttgaggcc
     1741 tatgaagggg tgctgcagct ggtggaggct cgcgggcgct acgaggagct ctgcatcgtc
     1801 atgtgtgtca tcccagccac catcagcaac aacgtccctg gcaccgactt cagcctgggc
     1861 tccgacactg ctgtaaatgc cgccatggag agctgtgacc gcatcaaaca gtctgcctcg
     1921 gggaccaagc gccgtgtgtt catcgtggag accatggggg gttactgtgg ctacctggcc
     1981 accgtgactg gcattgctgt gggggccgac gccgcctacg tcttcgagga ccctttcaac
     2041 atccacgact taaaggtcaa cgtggagcac atgacggaga agatgaagac agacattcag
     2101 aggggcctgg tgctgcggaa cgagaagtgc catgactact acaccacgga gttcctgtac
     2161 aacctgtact catcagaggg caagggcgtc ttcgactgca ggaccaatgt cctgggccac
     2221 ctgcagcagg gtggcgctcc aacccccttt gaccggaact atgggaccaa gctgggggtg
     2281 aaggccatgc tgtggttgtc ggagaagctg cgcgaggttt accgcaaggg acgggtgttc
     2341 gccaatgccc cagactcggc ctgcgtgatc ggcctgaaga agaaggcggt ggccttcagc
     2401 cccgtcactg agctcaagaa agacactgat ttcgagcacc gcatgccacg ggagcagtgg
     2461 tggctgagcc tgcggctcat gctgaagatg ctggcacaat accgcatcag tatggccgcc
     2521 tacgtgtcag gggagctgga gcacgtgacc cgccgcaccc tgagcatgga caagggcttc
     2581 tgcccaactt tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc
     2641 aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc
     2701 tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa
     2761 aatctctgat gttacattgc acaagataaa aatatatcat catgcctcct ctagaccagc
     2821 caggacagaa atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg
     2881 aaacggatga aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc
     2941 aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac
     3001 ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct
     3061 cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc
     3121 aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg
     3181 gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc
     3241 gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca
     3301 cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga
     3361 gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc
     3421 gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag
     3481 cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca
     3541 gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc
     3601 ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag
     3661 gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga
     3721 aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg
     3781 aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata
     3841 cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat
     3901 cagttggaag aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa
     3961 ccctcgagcc acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     4021 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4081 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4141 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4201 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4261 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4321 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4381 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4441 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4501 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4561 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4621 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4681 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     4741 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     4801 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     4861 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     4921 aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat
     4981 ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc
     5041 gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac
     5101 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag
     5161 cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg
     5221 ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc c
//
