LOCUS       sequence-375332-        4362 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 4362)
  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 4362)
  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-PAK2
FEATURES             Location/Qualifiers
     source          1..4362
                     /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(1673..1772)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1789..1808)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1790..1808)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1813..1829)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1813..1829)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2260..3051
                     /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      3144..3732
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3633..3652
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3886..3903
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4062..4089)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4181..4267)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4316..4338
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4330..4347
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4331..4347
                     /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 tgtctgataa cggagaactg
      121 gaagataagc ctccagcacc tcctgtgcga atgagcagca ccatctttag cactggaggc
      181 aaagaccctt tgtcagccaa tcacagtttg aaacctttgc cctctgttcc agaagagaaa
      241 aagcccaggc ataaaatcat ctccatattc tcaggcacag agaaaggaag taaaaagaag
      301 gaaaaggaac ggccagaaat ttctcctcca tctgattttg agcacaccat ccatgttggt
      361 tttgatgctg ttactggaga attcactggc atgccagaac agtgggctcg attactacag
      421 acctccaata tcaccaaact agagcaaaag aagaaccctc aggctgtgct ggatgtccta
      481 aagttctacg actccaacac agtgaagcag aaatatctga gctttactcc tcctgagaaa
      541 gatggctttc cttctggaac accagcactg aatgccaagg gaacagaagc acccgcagta
      601 gtgacagagg aggaggatga tgatgaagag actgctcctc ccgttattgc cccgcgaccg
      661 gatcatacga aatcaattta cacacggtct gtaattgacc ctgttcctgc accagttggt
      721 gattcacatg ttgatggtgc tgccaagtct ttagacaaac agaaaaagaa gactaagatg
      781 acagatgaag agattatgga gaaattaaga actatcgtga gcataggtga ccctaagaaa
      841 aaatatacaa gatatgaaaa aattggacaa agggcttctg gtacagtttt cactgctact
      901 gacgttgcac tgggacagga ggttgctatc aaacaaatta atttacagaa acagccaaag
      961 aaggaactga tcattaacga gattctggtg atgaaagaat tgaaaaatcc caacatcgtt
     1021 aactttttgg acagttacct ggtaggagat gaattgtttg tggtcatgga ataccttgct
     1081 ggggggtcac tcactgatgt ggtaacagaa acgtgcatgg atgaagcaca gattgctgct
     1141 gtatgcagag agtgtttaca ggcattggag tttttacatg ctaatcaagt gatccacaga
     1201 gacatcaaaa gtgacaatgt acttttggga atggaaggat ctgttaagct cactgacttt
     1261 ggtttctgtg cccagatcac ccctgagcag agcaaacgca gtaccatggt cggaacgcca
     1321 tactggatgg caccagaggt ggttacacgg aaagcttatg gccctaaagt cgacatatgg
     1381 tctctgggta tcatggctat tgagatggta gaaggagagc ctccatacct caatgaaaat
     1441 cccttgaggg ccttgtacct aatagcaact aatggaaccc cagaacttca gaatccagag
     1501 aaactttccc caatatttcg ggatttctta aatcgatgtt tggaaatgga tgtggaaaaa
     1561 aggggttcag ccaaagaatt attacagcat cctttcctga aactggccaa accgttatct
     1621 agcttgacac cactgatcat ggcagctaaa gaagcaatga agagtaaccg ttacccaact
     1681 ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag
     1741 gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag
     1801 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     1861 tgttacattg cacaagataa aaatatatca tcatgcctcc tctagaccag ccaggacaga
     1921 aatgcctcga cttcgctgct acccaaggtt gccgggtgac gcacaccgtg gaaacggatg
     1981 aaggcacgaa cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt
     2041 atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg
     2101 gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc
     2161 aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt
     2221 acgcagcagg gcagtcgccc taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc
     2281 gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg
     2341 ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat
     2401 attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc
     2461 aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa
     2521 gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg
     2581 caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc
     2641 gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt
     2701 ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat
     2761 gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg
     2821 cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc
     2881 gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct
     2941 agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa
     3001 gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata accctcgagc
     3061 cacccatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
     3121 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3181 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3241 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3301 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3361 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3421 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3481 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3541 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3601 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3661 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3721 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3781 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3841 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3901 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     3961 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     4021 aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga
     4081 tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
     4141 cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
     4201 cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt
     4261 ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag
     4321 tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cc
//
