LOCUS       sequence-490584-        4845 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 4845)
  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 4845)
  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-PLK2
FEATURES             Location/Qualifiers
     source          1..4845
                     /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"
     CDS             268..285
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(2156..2255)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2272..2291)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2273..2291)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2296..2312)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2296..2312)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2743..3534
                     /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      3627..4215
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4116..4135
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4369..4386
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4545..4572)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4664..4750)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4799..4821
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4813..4830
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4814..4830
                     /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 tggagctttt gcggactatc
      121 acctaccagc cagccgccag caccaaaatg tgcgagcagg cgctgggcaa gggttgcgga
      181 gcagactcga agaagaagcg gccgccgcag ccccccgagg aatcgcagcc acctcagtcc
      241 caggcgcaag tgcccccggc ggcccctcac caccatcacc accattcgca ctcggggccg
      301 gagatctcgc ggattatcgt cgaccccacg actgggaagc gctactgccg gggcaaagtg
      361 ctgggaaagg gtggctttgc aaaatgttac gagatgacag atttgacaaa taacaaagtc
      421 tacgccgcaa aaattattcc tcacagcaga gtagctaaac ctcatcaaag ggaaaagatt
      481 gacaaagaaa tagagcttca cagaattctt catcataagc atgtagtgca gttttaccac
      541 tacttcgagg acaaagaaaa catttacatt ctcttggaat actgcagtag aaggtcaatg
      601 gctcatattt tgaaagcaag aaaggtgttg acagagccag aagttcgata ctacctcagg
      661 cagattgtgt ctggactgaa ataccttcat gaacaagaaa tcttgcacag agatctcaaa
      721 ctagggaact tttttattaa tgaagccatg gaactaaaag ttggggactt cggtctggca
      781 gccaggctag aacccttgga acacagaagg agaacgatat gtggtacccc aaattatctc
      841 tctcctgaag tcctcaacaa acaaggacat ggctgtgaat cagacatttg ggccctgggc
      901 tgtgtaatgt atacaatgtt actagggagg cccccatttg aaactacaaa tctcaaagaa
      961 acttataggt gcataaggga agcaaggtat acaatgccgt cctcattgct ggctcctgcc
     1021 aagcacttaa ttgctagtat gttgtccaaa aacccagagg atcgtcccag tttggatgac
     1081 atcattcgac atgacttttt tttgcagggc ttcactccgg acagactgtc ttctagctgt
     1141 tgtcatacag ttccagattt ccacttatca agcccagcta agaatttctt taagaaagca
     1201 gctgctgctc tttttggtgg caaaaaagac aaagcaagat atattgacac acataataga
     1261 gtgtctaaag aagatgaaga catctacaag cttaggcatg atttgaaaaa gacttcaata
     1321 actcagcaac ccagcaaaca caggacagat gaggagctcc agccacctac caccacagtt
     1381 gccaggtctg gaacacccgc agtagaaaac aagcagcaga ttggggatgc tattcggatg
     1441 atagtcagag ggactcttgg cagctgtagc agcagcagtg aatgccttga agacagtacc
     1501 atgggaagtg ttgcagacac agtggcaagg gttcttcggg gatgtctgga aaacatgccg
     1561 gaagctgatt gcattcccaa agagcagctg agcacatcat ttcagtgggt caccaaatgg
     1621 gttgattact ctaacaaata tggctttggg taccagctct cagaccacac cgtcggtgtc
     1681 cttttcaaca atggtgctca catgagcctc cttccagaca aaaaaacagt tcactattac
     1741 gcagagcttg gccaatgctc agttttccca gcaacagatg ctcctgagca atttattagt
     1801 caagtgacgg tgctgaaata cttttctcat tacatggagg agaacctcat ggatggtgga
     1861 gatctgccta gtgttactga tattcgaaga cctcggctct acctccttca gtggctaaaa
     1921 tctgataagg ccctaatgat gctctttaat gatggcacct ttcaggtgaa tttctaccat
     1981 gatcatacaa aaatcatcat ctgtagccaa aatgaagaat accttctcac ctacatcaat
     2041 gaggatagga tatctacaac tttcaggctg acaactctgc tgatgtctgg ctgttcatca
     2101 gaattaaaaa atcgaatgga atatgccctg aacatgctct tacaaagatg taactgccca
     2161 actttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa
     2221 caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt
     2281 gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc
     2341 tgatgttaca ttgcacaaga taaaaatata tcatcatgcc tcctctagac cagccaggac
     2401 agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg
     2461 atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag
     2521 cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca
     2581 gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca
     2641 tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat
     2701 gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc
     2761 gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg
     2821 acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt
     2881 gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg
     2941 atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta
     3001 gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa
     3061 ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg
     3121 atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta
     3181 ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta
     3241 aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta
     3301 gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat
     3361 gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa
     3421 gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg
     3481 gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg
     3541 agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac
     3601 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     3661 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     3721 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     3781 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     3841 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     3901 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3961 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4021 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4081 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4141 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4201 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4261 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     4321 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     4381 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     4441 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     4501 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca
     4561 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
     4621 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
     4681 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc
     4741 gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc
     4801 cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccc
//
