LOCUS       sequence-305008-        5529 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 5529)
  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 5529)
  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-PRKD1
FEATURES             Location/Qualifiers
     source          1..5529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(919..946)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1038..1124)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1173..1195
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1187..1204
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1188..1204
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1220..1319
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4059..4158)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4175..4194)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4176..4194)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4199..4215)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4199..4215)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4646..5437
                     /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"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga
      901 agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc
      961 ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc
     1021 aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata
     1081 aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc
     1141 tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc
     1201 cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga cctgttcgtt
     1261 gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa aagttggcac
     1321 catgagcgcc cctccggtcc tgcggccgcc cagtccgctg ctgcccgtgg cggcggcagc
     1381 tgccgcagcg gccgccgcac tggtcccagg gtccgggccc gggcccgcgc cgttcttggc
     1441 tcctgtcgcg gccccggtcg ggggcatctc gttccatctg cagatcggcc tgagccgtga
     1501 gccggtgctg ctgctgcagg actcgtccgg ggactacagc ctggcgcacg tccgcgagat
     1561 ggcttgctcc attgtcgacc agaagttccc tgaatgtggt ttctacggaa tgtatgataa
     1621 gatcctgctt tttcgccatg accctacctc tgaaaacatc cttcagctgg tgaaagcggc
     1681 cagtgatatc caggaaggcg atcttattga agtggtcttg tcagcttccg ccacctttga
     1741 agactttcag attcgtcccc acgctctctt tgttcattca tacagagctc cagctttctg
     1801 tgatcactgt ggagaaatgc tgtgggggct ggtacgtcaa ggtcttaaat gtgaagggtg
     1861 tggtctgaat taccataaga gatgtgcatt taaaataccc aacaattgca gcggtgtgag
     1921 gcggagaagg ctctcaaacg tttccctcac tggggtcagc accatccgca catcatctgc
     1981 tgaactctct acaagtgccc ctgatgagcc ccttctgcaa aaatcaccat cagagtcgtt
     2041 tattggtcga gagaagaggt caaattctca atcatacatt ggacgaccaa ttcaccttga
     2101 caagattttg atgtctaaag ttaaagtgcc gcacacattt gtcatccact cctacacccg
     2161 gcccacagtg tgccagtact gcaagaagct tctgaagggg cttttcaggc agggcttgca
     2221 gtgcaaagat tgcagattca actgccataa acgttgtgca ccgaaagtac caaacaactg
     2281 ccttggcgaa gtgaccatta atggagattt gcttagccct ggggcagagt ctgatgtggt
     2341 catggaagaa gggagtgatg acaatgatag tgaaaggaac agtgggctca tggatgatat
     2401 ggaagaagca atggtccaag atgcagagat ggcaatggca gagtgccaga acgacagtgg
     2461 cgagatgcaa gatccagacc cagaccacga ggacgccaac agaaccatca gtccatcaac
     2521 aagcaacaat atcccactca tgagggtagt gcagtctgtc aaacacacga agaggaaaag
     2581 cagcacagtc atgaaagaag gatggatggt ccactacacc agcaaggaca cgctgcggaa
     2641 acggcactat tggagattgg atagcaaatg tattaccctc tttcagaatg acacaggaag
     2701 caggtactac aaggaaattc ctttatctga aattttgtct ctggaaccag taaaaacttc
     2761 agctttaatt cctaatgggg ccaatcctca ttgtttcgaa atcactacgg caaatgtagt
     2821 gtattatgtg ggagaaaatg tggtcaatcc ttccagccca tcaccaaata acagtgttct
     2881 caccagtggc gttggtgcag atgtggccag gatgtgggag atagccatcc agcatgccct
     2941 tatgcccgtc attcccaagg gctcctccgt gggtacagga accaacttgc acagagatat
     3001 ctctgtgagt atttcagtat caaattgcca gattcaagaa aatgtggaca tcagcacagt
     3061 atatcagatt tttcctgatg aagtactggg ttctggacag tttggaattg tttatggagg
     3121 aaaacatcgt aaaacaggaa gagatgtagc tattaaaatc attgacaaat tacgatttcc
     3181 aacaaaacaa gaaagccagc ttcgtaatga ggttgcaatt ctacagaacc ttcatcaccc
     3241 tggtgttgta aatttggagt gtatgtttga gacgcctgaa agagtgtttg ttgttatgga
     3301 aaaactccat ggagacatgc tggaaatgat cttgtcaagt gaaaagggca ggttgccaga
     3361 gcacataacg aagtttttaa ttactcagat actcgtggct ttgcggcacc ttcattttaa
     3421 aaatatcgtt cactgtgacc tcaaaccaga aaatgtgttg ctagcctcag ctgatccttt
     3481 tcctcaggtg aaactttgtg attttggttt tgcccggatc atcggagaga agtctttccg
     3541 gaggtcagtg gtgggtaccc ccgcttacct ggctcctgag gtcctaagga acaagggcta
     3601 caatcgctct ctagacatgt ggtctgttgg ggtcatcatc tatgtaagcc taagcggcac
     3661 attcccattt aatgaagatg aagacataca cgaccaaatt cagaatgcag ctttcatgta
     3721 tccaccaaat ccctggaagg aaatatctca tgaagccatt gatcttatca acaatttgct
     3781 gcaagtaaaa atgagaaagc gctacagtgt ggataagacc ttgagccacc cttggctaca
     3841 ggactatcag acctggttag atttgcgaga gctggaatgc aaaatcgggg agcgctacat
     3901 cacccatgaa agtgatgacc tgaggtggga gaagtatgca ggcgagcagg ggctgcagta
     3961 ccccacacac ctgatcaatc caagtgctag ccacagtgac actcctgaga ctgaagaaac
     4021 agaaatgaaa gccctcggtg agcgtgtcag catcctcttg ccaactttct tgtacaaagt
     4081 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     4141 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     4201 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     4261 agataaaaat atatcatcat gcctcctcta gaccagccag gacagaaatg cctcgacttc
     4321 gctgctaccc aaggttgccg ggtgacgcac accgtggaaa cggatgaagg cacgaaccca
     4381 gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa
     4441 ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc
     4501 ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg
     4561 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag
     4621 tcgccctaaa acaaagttaa acattatgag ggaagcggtg atcgccgaag tatcgactca
     4681 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
     4741 tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt
     4801 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga
     4861 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
     4921 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
     4981 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc
     5041 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
     5101 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
     5161 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg
     5221 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc
     5281 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
     5341 tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta
     5401 cgtgaaaggc gagatcacca aggtagtcgg caaataaccc tcgagccacc catgaccaaa
     5461 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5521 aggatcttc
//
