LOCUS       sequence-297880-        4350 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 4350)
  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 4350)
  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-PFKFB3
FEATURES             Location/Qualifiers
     source          1..4350
                     /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(2229..2328)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2345..2364)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2346..2364)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2369..2385)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2369..2385)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2816..3607
                     /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      3700..4288
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4189..4208
                     /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 ccgttggaac tgacgcagag ccgagtgcag aagatctggg tgcccgtgga
      721 ccacaggccc tcgttgccca gatcctgtgg gccaaagctg accaactccc ccaccgtcat
      781 cgtcatggtg ggcctccccg cccggggcaa gacctacatc tccaagaagc tgactcgcta
      841 cctcaactgg attggcgtcc ccacaaaagt gttcaacgtc ggggagtatc gccgggaggc
      901 tgtgaagcag tacagctcct acaacttctt ccgccccgac aatgaggaag ccatgaaagt
      961 ccggaagcaa tgtgccttag ctgccttgag agatgtcaaa agctacctgg cgaaagaagg
     1021 gggacaaatt gcggttttcg atgccaccaa tactactaga gagaggagac acatgatcct
     1081 tcattttgcc aaagaaaatg actttaaggc gtttttcatc gagtcggtgt gcgacgaccc
     1141 tacagttgtg gcctccaata tcatggaagt taaaatctcc agcccggatt acaaagactg
     1201 caactcggca gaagccatgg acgacttcat gaagaggatc agttgctatg aagccagcta
     1261 ccagcccctc gaccccgaca aatgcgacag ggacttgtcg ctgatcaagg tgattgacgt
     1321 gggccggagg ttcctggtga accgggtgca ggaccacatc cagagccgca tcgtgtacta
     1381 cctgatgaac atccacgtgc agccgcgtac catctacctg tgccggcacg gcgagaacga
     1441 gcacaacctc cagggccgca tcgggggcga ctcaggcctg tccagccggg gcaagaagtt
     1501 tgccagtgct ctgagcaagt tcgtggagga gcagaacctg aaggacctgc gcgtgtggac
     1561 cagccagctg aagagcacca tccagacggc cgaggcgctg cggctgccct acgagcagtg
     1621 gaaggcgctc aatgagatcg acgcgggcgt ctgtgaggag ctgacctacg aggagatcag
     1681 ggacacctac cctgaggagt atgcgctgcg ggagcaggac aagtactatt accgctaccc
     1741 caccggggag tcctaccagg acctggtcca gcgcttggag ccagtgatca tggagctgga
     1801 gcggcaggag aatgtgctgg tcatctgcca ccaggccgtc ctgcgctgcc tgcttgccta
     1861 cttcctggat aagagtgcag aggagatgcc ctacctgaaa tgccctcttc acaccgtcct
     1921 gaaactgacg cctgtcgctt atggctgccg tgtggaatcc atctacctga acgtggagtc
     1981 cgtctgcaca caccgggaga ggtcagagga tgcaaagaag ggacctaacc cgctcatgag
     2041 acgcaatagt gtcaccccgc tagccagccc cgaacccacc aaaaagcctc gcatcaacag
     2101 ctttgaggag catgtggcct ccacctcggc cgccctgccc agctgcctgc ccccggaggt
     2161 gcccacgcag ctgcctggac aaaacatgaa aggctcccgg agcagcgctg actcctccag
     2221 gaaacactgc ccaactttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     2281 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     2341 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     2401 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gcctcctcta
     2461 gaccagccag gacagaaatg cctcgacttc gctgctaccc aaggttgccg ggtgacgcac
     2521 accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct
     2581 gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg
     2641 tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc
     2701 tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg
     2761 gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa acattatgag
     2821 ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg
     2881 ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct
     2941 gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac
     3001 gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat
     3061 tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc
     3121 agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt
     3181 cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag
     3241 cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct
     3301 atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga
     3361 tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat
     3421 cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc
     3481 cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg
     3541 cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg
     3601 caaataaccc tcgagccacc catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     3661 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3721 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3781 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3841 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3901 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3961 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4021 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4081 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4141 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4201 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4261 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4321 ggccttttgc tggccttttg ctcacatgtt
//
