LOCUS       sequence-343698-        5142 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 5142)
  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 5142)
  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-PFKP
FEATURES             Location/Qualifiers
     source          1..5142
                     /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"
     protein_bind    complement(2453..2552)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2569..2588)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2570..2588)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2593..2609)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2593..2609)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3040..3831
                     /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      3924..4512
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4413..4432
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4666..4683
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4842..4869)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4961..5047)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5096..5118
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5110..5127
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5111..5127
                     /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 tggacgcgga cgactcccgg
      121 gcccccaagg gctccttgcg gaagttcctg gagcacctct ccggggccgg caaggccatc
      181 ggcgtgctga ccagcggcgg ggatgctcaa ggtatgaacg ctgccgtccg tgccgtggtg
      241 cgcatgggta tctacgtggg ggccaaggtg tacttcatct acgagggcta ccagggcatg
      301 gtggacggag gctcaaacat cgcagaggcc gactgggaga gtgtctccag catcctgcaa
      361 gtgggcggga cgatcattgg cagtgcgcgg tgccaggcct tccgcacgcg ggaaggccgc
      421 ctgaaggctg cttgcaacct gctgcagcgc ggcatcacca acctgtgtgt gatcggcggg
      481 gacgggagcc tcaccggggc caacctcttc cggaaggagt ggagtgggct gctggaggag
      541 ctggccagga acggccagat cgataaggag gccgtgcaga agtacgccta cctcaacgtg
      601 gtgggcatgg tgggctccat cgacaatgat ttctgcggca ccgacatgac catcggcacg
      661 gactccgccc tgcacaggat catcgaggtc gtcgacgcca tcatgaccac ggcccagagc
      721 caccagagga ccttcgttct ggaggtgatg ggacgacact gtgggtacct ggccctggtg
      781 agtgccttgg cctgcggtgc ggactgggtg ttccttccag aatctccacc agaggaaggc
      841 tgggaggagc agatgtgtgt caaactctcg gagaaccgtg cccggaaaaa aaggctgaat
      901 attattattg tggctgaagg agcaattgat acccaaaata aacccatcac ctctgagaaa
      961 atcaaagagc ttgtcgtcac gcagctgggc tatgacacac gtgtgaccat cctcgggcac
     1021 gtgcagagag gagggacccc ttcggcattc gacaggatct tggccagccg catgggagtg
     1081 gaggcagtca tcgccttgct agaggccacc ccggacaccc cagcttgcgt cgtgtcactg
     1141 aacgggaacc acgccgtgcg cctgccgctg atggagtgcg tgcagatgac tcaggatgtg
     1201 cagaaggcga tggacgagag gagatttcaa gatgcggttc gactccgagg gaggagcttt
     1261 gcgggcaacc tgaacaccta caagcgactt gccatcaagc tgccggatga tcagatccca
     1321 aagaccaatt gcaacgtagc tgtcatcaac gtgggggcac ccgcggctgg gatgaacgcg
     1381 gccgtacgct cagctgtgcg cgtgggcatt gccgacggcc acaggatgct cgccatctat
     1441 gatggctttg acggcttcgc caagggccag atcaaagaaa tcggctggac agatgtcggg
     1501 ggctggaccg gccaaggagg ctccattctt gggacaaaac gcgttctccc ggggaagtac
     1561 ttggaagaga tcgccacaca gatgcgcacg cacagcatca acgcgctgct gatcatcggt
     1621 ggattcgagg cctacctggg actcctggag ctgtcagccg cccgggagaa gcacgaggag
     1681 ttctgtgtcc ccatggtcat ggttcccgct actgtgtcca acaatgtgcc gggttccgat
     1741 ttcagcatcg gggcagacac cgccctgaac actatcaccg acacctgcga ccgcatcaag
     1801 cagtccgcca gcggaaccaa gcggcgcgtg ttcatcatcg agaccatggg cggctactgt
     1861 ggctacctgg ccaacatggg ggggctcgcg gctggagctg atgccgcata cattttcgaa
     1921 gagcccttcg acatcaggga tctgcagtcc aacgtggagc acctgacgga gaaaatgaag
     1981 accaccatcc agagaggcct tgtgctcaga aatgagagct gcagtgaaaa ctacaccacc
     2041 gacttcattt accagctgta ttcagaagag ggcaaaggcg tgtttgactg caggaagaac
     2101 gtgctgggtc acatgcagca gggtggggca ccctctccat ttgatagaaa ctttggaacc
     2161 aaaatctctg ccagagctat ggagtggatc actgcaaaac tcaaggaggc ccggggcaga
     2221 ggaaaaaaat ttaccaccga tgattccatt tgtgtgctgg gaataagcaa aagaaacgtt
     2281 atttttcaac ctgtggcaga gctgaagaag caaacggatt ttgagcacag gattcccaaa
     2341 gaacagtggt ggctcaagct acggcccctc atgaaaatcc tggccaagta caaggccagc
     2401 tatgacgtgt cggactcagg ccagctggaa catgtgcagc cctggagtgt ctgcccaact
     2461 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     2521 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag
     2581 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     2641 tgttacattg cacaagataa aaatatatca tcatgcctcc tctagaccag ccaggacaga
     2701 aatgcctcga cttcgctgct acccaaggtt gccgggtgac gcacaccgtg gaaacggatg
     2761 aaggcacgaa cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt
     2821 atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg
     2881 gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc
     2941 aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt
     3001 acgcagcagg gcagtcgccc taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc
     3061 gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg
     3121 ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat
     3181 attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc
     3241 aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa
     3301 gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg
     3361 caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc
     3421 gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt
     3481 ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat
     3541 gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg
     3601 cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc
     3661 gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct
     3721 agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa
     3781 gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata accctcgagc
     3841 cacccatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     3901 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3961 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4021 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4081 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     4141 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     4201 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     4261 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     4321 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     4381 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     4441 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4501 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4561 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4621 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     4681 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     4741 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     4801 aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga
     4861 tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
     4921 cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     4981 cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt
     5041 ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag
     5101 tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cc
//
