LOCUS       sequence-345435-        4911 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 4911)
  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 4911)
  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-PRKCQ
FEATURES             Location/Qualifiers
     source          1..4911
                     /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(2222..2321)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2338..2357)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2339..2357)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2362..2378)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2362..2378)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2809..3600
                     /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      3693..4281
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4182..4201
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4435..4452
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4611..4638)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4730..4816)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4865..4887
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4879..4896
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4880..4896
                     /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 ccatgtcgcc atttcttcgg
      121 attggcttgt ccaactttga ctgcgggtcc tgccagtctt gtcagggcga ggctgttaac
      181 ccttactgtg ctgtgctcgt caaagagtat gtcgaatcag agaacgggca gatgtatatc
      241 cagaaaaagc ctaccatgta cccaccctgg gacagcactt ttgatgccca tatcaacaag
      301 ggaagagtca tgcagatcat tgtgaaaggc aaaaacgtgg acctcatctc tgaaaccacc
      361 gtggagctct actcgctggc tgagaggtgc aggaagaaca acgggaagac agaaatatgg
      421 ttagagctga aacctcaagg ccgaatgcta atgaatgcaa gatactttct ggaaatgagt
      481 gacacaaagg acatgaatga atttgagacg gaaggcttct ttgctttgca tcagcgccgg
      541 ggtgccatca agcaggcaaa ggtccaccac gtcaagtgcc acgagttcac tgccaccttc
      601 ttcccacagc ccacattttg ctctgtctgc cacgagtttg tctggggcct gaacaaacag
      661 ggctaccagt gccgacaatg caatgcagca attcacaaga agtgtattga taaagttata
      721 gcaaagtgca caggatcagc tatcaatagc cgagaaacca tgttccacaa ggagagattc
      781 aaaattgaca tgccacacag atttaaagtc tacaattaca agagcccgac cttctgtgaa
      841 cactgtggga ccctgctgtg gggactggca cggcaaggac tcaagtgtga tgcatgtggc
      901 atgaatgtgc atcatagatg ccagacaaag gtggccaacc tttgtggcat aaaccagaag
      961 ctaatggctg aagcgctggc catgattgag agcactcaac aggctcgctg cttaagagat
     1021 actgaacaga tcttcagaga aggtccggtt gaaattggtc tcccatgctc catcaaaaat
     1081 gaagcaaggc cgccatgttt accgacaccg ggaaaaagag agcctcaggg catttcctgg
     1141 gagtctccgt tggatgaggt ggataaaatg tgccatcttc cagaacctga actgaacaaa
     1201 gaaagaccat ctctgcagat taaactaaaa attgaggatt ttatcttgca caaaatgttg
     1261 gggaaaggaa gttttggcaa ggtcttcctg gcagaattca agaaaaccaa tcaatttttc
     1321 gcaataaagg ccttaaagaa agatgtggtc ttgatggacg atgatgttga gtgcacgatg
     1381 gtagagaaga gagttctttc cttggcctgg gagcatccgt ttctgacgca catgttttgt
     1441 acattccaga ccaaggaaaa cctctttttt gtgatggagt acctcaacgg aggggactta
     1501 atgtaccaca tccaaagctg ccacaagttc gacctttcca gagcgacgtt ttatgctgct
     1561 gaaatcattc ttggtctgca gttccttcat tccaaaggaa tagtctacag ggacctgaag
     1621 ctagataaca tcctgttaga caaagatgga catatcaaga ttgcggattt tggaatgtgc
     1681 aaggagaaca tgttaggaga tgccaagacg aataccttct gtgggacacc tgactacatc
     1741 gccccagaga tcttgctggg tcagaaatac aaccactctg tggactggtg gtccttcggg
     1801 gttctccttt atgaaatgct gattggtcag tcgcctttcc acgggcagga tgaggaggag
     1861 ctcttccact ccatccgcat ggacaatccc ttttacccac ggtggctgga gaaggaagca
     1921 aaggaccttc tggtgaagct cttcgtgaga gaacctgaga agaggctggg cgtgagggga
     1981 gacatccgcc agcacccttt gtttcgggag atcaactggg aggaacttga acggaaggag
     2041 attgacccac cgttccggcc gaaagtgaaa tcaccatttg actgcagcaa tttcgacaaa
     2101 gaattcttaa acgagaagcc ccggctgtca tttgccgaca gagcactgat caacagcatg
     2161 gaccagaata tgttcaggaa cttttccttc atgaaccccg ggatggagcg gctgatatcc
     2221 ttgccaactt tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc
     2281 aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc
     2341 tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa
     2401 aatctctgat gttacattgc acaagataaa aatatatcat catgcctcct ctagaccagc
     2461 caggacagaa atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg
     2521 aaacggatga aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc
     2581 aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac
     2641 ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct
     2701 cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc
     2761 aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg
     2821 gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc
     2881 gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca
     2941 cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga
     3001 gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc
     3061 gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag
     3121 cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca
     3181 gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc
     3241 ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag
     3301 gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga
     3361 aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg
     3421 aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata
     3481 cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat
     3541 cagttggaag aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa
     3601 ccctcgagcc acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg
     3661 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3721 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3781 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     3841 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     3901 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     3961 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4021 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4081 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4141 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4201 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4261 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4321 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     4381 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     4441 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     4501 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     4561 aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat
     4621 ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc
     4681 gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac
     4741 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag
     4801 cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg
     4861 ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc c
//
