LOCUS       sequence-459892-        4462 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 4462)
  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 4462)
  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-HIPK1
FEATURES             Location/Qualifiers
     source          1..4462
                     /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(1773..1872)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1889..1908)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1890..1908)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1913..1929)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1913..1929)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2360..3151
                     /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      3244..3832
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3733..3752
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3986..4003
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4162..4189)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4281..4367)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4416..4438
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4430..4447
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4431..4447
                     /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 tggttttgat gtttcagatt
      121 cgttatattt cacaaacaca aggcttgcca gctgaatatc ttctcagtgc cggaacaaaa
      181 acaaccaggt ttttcaacag agatcctaat ttggggtacc cactgtggag gcttaagaca
      241 cctgaagaac atgaactgga gactggaata aaatcaaaag aagctcggaa gtacattttt
      301 aattgcttag atgacatggc tcaggtgaat atgtctacgg acctggaggg aacagacatg
      361 ttggcagaga aggcagaccg aagagaatac attgatctgt taaagaaaat gctcacaatt
      421 gatgcagata agagaattac ccctctaaaa actcttaacc atcagtttgt gacaatgact
      481 caccttttgg attttccaca tagcaatcat gttaagtctt gttttcagaa catggagatc
      541 tgcaagcgga gggttcacat gtatgataca gtgagtcaga tcaagagtcc cttcactaca
      601 catgttgccc caaatacaag cacaaatcta accatgagct tcagcaatca gctcaataca
      661 gtgcacaatc aggccagtgt tctagcttcc agttctactg cagcagctgc tactctttct
      721 ctggctaatt cagatgtctc actactaaac taccagtcag ctttgtacca catcttctta
      781 taattccttg gtccctgtcc aagatcagca tcagcccatc atcattccag atactcccag
      841 ccctcctgtg agtgtcatca ctatccgaag tgacactgat gaggaagagg acaacaaata
      901 caagcccagt agctctggac tgaagccaag gtctaatgtc atcagttatg tcactgtcaa
      961 tgattctcct gactctgact cttctttgag cagcccttat tccactgata ccctgagtgc
     1021 tctccgaggc aatagtggat ccgttttgga ggggcctggc agagttgtgg cagatggcac
     1081 tggcacccgc actatcattg tgcctccact gaaaactcag cttggtgact gcactgtagc
     1141 aacccaggcc tcaggtctcc tgagcaataa gactaagcca gtcgcttcag tgagtgggca
     1201 gtcatctgga tgctgtatca cccccacagg gtatcgagct caacgcgggg ggaccagtgc
     1261 agcacaacca ctcaatctta gccagaacca gcagtcatcg gtggctccaa cctcacagga
     1321 gagaagcagc aacccagccc cccgcaggca gcaggcgttt gtggcccctc tctcccaagc
     1381 cccctacacc ttccagcatg gcagcccgct acactcgaca gggcacccac accttgcccc
     1441 ggcccctgct cacctgccaa gccaggctca tctgtatacg tatgctgccc cgacttctgc
     1501 tgctgcactg ggctcaacca gctccattgc tcatcttttc tccccacagg gttcctcaag
     1561 gcatgctgca gcctatacca ctcaccctag cactttggtg caccaggtcc ctgtcagtgt
     1621 tgggcccagc ctcctcactt ctgccagcgt ggcccctgct cagtaccaac accagtttgc
     1681 cacccaatcc tacattgggt cttcccgagg ctcaacaatt tacactggat acccgctgag
     1741 tcctaccaag atcagccagt attcctactt atacccaact ttcttgtaca aagttggcat
     1801 tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa
     1861 aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc
     1921 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     1981 aaatatatca tcatgcctcc tctagaccag ccaggacaga aatgcctcga cttcgctgct
     2041 acccaaggtt gccgggtgac gcacaccgtg gaaacggatg aaggcacgaa cccagtggac
     2101 ataagcctgt tcggttcgta agctgtaatg caagtagcgt atgcgctcac gcaactggtc
     2161 cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta
     2221 tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc
     2281 cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg gcagtcgccc
     2341 taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc
     2401 agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta
     2461 cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc tggttacggt
     2521 gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc
     2581 ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga
     2641 cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg
     2701 caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt
     2761 gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt
     2821 tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa
     2881 ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg
     2941 gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga
     3001 gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca
     3061 agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa
     3121 aggcgagatc accaaggtag tcggcaaata accctcgagc cacccatgac caaaatccct
     3181 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3241 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3301 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3361 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3421 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3481 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3541 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3601 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3661 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3721 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3781 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3841 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3901 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3961 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     4021 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     4081 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca
     4141 ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga
     4201 tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg
     4261 ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag
     4321 ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt
     4381 ccctactctc gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac
     4441 ggccagtctt aagctcgggc cc
//
