LOCUS       sequence-450654-        6189 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 6189)
  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 6189)
  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-JAK2
FEATURES             Location/Qualifiers
     source          1..6189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(637..736)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(753..772)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(754..772)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(777..793)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(777..793)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1224..2015
                     /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      2108..2696
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2597..2616
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2850..2867
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3026..3053)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3145..3231)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3280..3302
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3294..3311
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3295..3311
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3327..3426
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ggcgtaatct aaaattaatt atggaatatt taccatatgg aagtttacga gactatcttc
       61 aaaaacataa agaacggata gatcacataa aacttctgca gtacacatct cagatatgca
      121 agggtatgga gtatcttggt acaaaaaggt atatccacag ggatctggca acgagaaata
      181 tattggtgga gaacgagaac agagttaaaa ttggagattt tgggttaacc aaagtcttgc
      241 cacaagacaa agaatactat aaagtaaaag aacctggtga aagtcccata ttctggtatg
      301 ctccagaatc actgacagag agcaagtttt ctgtggcctc agatgtttgg agctttggag
      361 tggttctgta tgaacttttc acatacattg agaagagtaa aagtccacca gcggaattta
      421 tgcgtatgat tggcaatgac aaacaaggac agatgatcgt gttccatttg atagaacttt
      481 tgaagaataa tggaagatta ccaagaccag atggatgccc agatgagatc tatatgatca
      541 tgacagaatg ctggaacaat aatgtaaatc aacgcccctc ctttagggat ctagctcttc
      601 gagtggatca aataagggat aacatggctg gatgatgccc aactttcttg tacaaagttg
      661 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
      721 ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca
      781 tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
      841 ataaaaatat atcatcatgc ctcctctaga ccagccagga cagaaatgcc tcgacttcgc
      901 tgctacccaa ggttgccggg tgacgcacac cgtggaaacg gatgaaggca cgaacccagt
      961 ggacataagc ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact
     1021 ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt
     1081 gttatgactg tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt
     1141 acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc
     1201 gccctaaaac aaagttaaac attatgaggg aagcggtgat cgccgaagta tcgactcaac
     1261 tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt
     1321 tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta
     1381 cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa
     1441 cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc
     1501 acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc
     1561 agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta
     1621 tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac
     1681 tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat
     1741 ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca
     1801 tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa
     1861 tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg
     1921 gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg
     1981 tgaaaggcga gatcaccaag gtagtcggca aataaccctc gagccaccca tgaccaaaat
     2041 cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     2101 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     2161 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     2221 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     2281 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     2341 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     2401 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     2461 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     2521 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     2581 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     2641 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     2701 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     2761 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     2821 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     2881 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     2941 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta
     3001 gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt
     3061 ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac
     3121 aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca
     3181 acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc
     3241 agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgttgtaaaa
     3301 cgacggccag tcttaagctc gggccccaaa taatgatttt attttgactg atagtgacct
     3361 gttcgttgca acaaattgat gagcaatgct tttttataat gccaactttg tacaaaaaag
     3421 ttggcatggg aatggcctgc cttacgatga cagaaatgga gggaacatcc acctcttcta
     3481 tatatcagaa tggtgatatt tctggaaatg ccaattctat gaagcaaata gatccagttc
     3541 ttcaggtgta tctttaccat tcccttggga aatctgaggc agattatctg acctttccat
     3601 ctggggagta tgttgcagaa gaaatctgta ttgctgcttc taaagcttgt ggtatcacac
     3661 ctgtgtatca taatatgttt gctttaatga gtgaaacaga aaggatctgg tatccaccca
     3721 accatgtctt ccatatagat gagtcaacca ggcataatgt actctacaga ataagatttt
     3781 actttcctcg ttggtattgc agtggcagca acagagccta tcggcatgga atatctcgag
     3841 gtgctgaagc tcctcttctt gatgactttg tcatgtctta cctctttgct cagtggcggc
     3901 atgattttgt gcacggatgg ataaaagtac ctgtgactca tgaaacacag gaagaatgtc
     3961 ttgggatggc agtgttagat atgatgagaa tagccaaaga aaacgatcaa accccactgg
     4021 ccatctataa ctctatcagc tacaagacat tcttaccaaa atgtattcga gcaaagatcc
     4081 aagactatca tattttgaca aggaagcgaa taaggtacag atttcgcaga tttattcagc
     4141 aattcagcca atgcaaagcc actgccagaa acttgaaact taagtatctt ataaatctgg
     4201 aaactctgca gtctgccttc tacacagaga aatttgaagt aaaagaacct ggaagtggtc
     4261 cttcaggtga ggagattttt gcaaccatta taataactgg aaacggtgga attcagtggt
     4321 caagagggaa acataaagaa agtgagacac tgacagaaca ggatttacag ttatattgcg
     4381 attttcctaa tattattgat gtcagtatta agcaagcaaa ccaagagggt tcaaatgaaa
     4441 gccgagttgt aactatccat aagcaagatg gtaaaaatct ggaaattgaa cttagctcat
     4501 taagggaagc tttgtctttc gtgtcattaa ttgatggata ttatagatta actgcagatg
     4561 cacatcatta cctctgtaaa gaagtagcac ctccagccgt gcttgaaaat atacaaagca
     4621 actgtcatgg cccaatttcg atggattttg ccattagtaa actgaagaaa gcaggtaatc
     4681 agactggact gtatgtactt cgatgcagtc ctaaggactt taataaatat tttttgactt
     4741 ttgctgtcga gcgagaaaat gtcattgaat ataaacactg tttgattaca aaaaatgaga
     4801 atgaagagta caacctcagt gggacaaaga agaacttcag cagtcttaaa gatcttttga
     4861 attgttacca gatggaaact gttcgctcag acaatataat tttccagttt actaaatgct
     4921 gtcccccaaa gccaaaagat aaatcaaacc ttctagtctt cagaacgaat ggtgtttctg
     4981 atgtaccaac ctcaccaaca ttacagaggc ctactcatat gaaccaaatg gtgtttcaca
     5041 aaatcagaaa tgaagatttg atatttaatg aaagccttgg ccaaggcact tttacaaaga
     5101 tttttaaagg cgtacgaaga gaagtaggag actacggtca actgcatgaa acagaagttc
     5161 ttttaaaagt tctggataaa gcacacagaa actattcaga gtctttcttt gaagcagcaa
     5221 gtatgatgag caagctttct cacaagcatt tggttttaaa ttatggagta tgtgtctgtg
     5281 gagacgagaa tattctggtt caggagtttg taaaatttgg atcactagat acatatctga
     5341 aaaagaataa aaattgtata aatatattat ggaaacttga agttgctaaa cagttggcat
     5401 gggccatgca ttttctagaa gaaaacaccc ttattcatgg gaatgtatgt gccaaaaata
     5461 ttctgcttat cagagaagaa gacaggaaga caggaaatcc tcctttcatc aaacttagtg
     5521 atcctggcat tagtattaca gttttgccaa aggacattct tcaggagaga ataccatggg
     5581 taccacctga atgcattgaa aatcctaaaa atttaaattt ggcaacagac aaatggagtt
     5641 ttggtaccac tttgtgggaa atctgcagtg gaggagataa acctctaagt gctctggatt
     5701 ctcaaagaaa gctacaattt tatgaagata ggcatcagct tcctgcacca aagtgggcag
     5761 aattagcaaa ccttataaat aattgtatgg attatgaacc agatttcagg ccttctttca
     5821 gagccatcat acgagatctt aacagtttgt ttactccaga ttatgaacta ttaacagaaa
     5881 atgacatgtt accaaatatg aggataggtg ccctagggtt ttctggtgcc tttgaagacc
     5941 gggatcctac acagtttgaa gagagacatt tgaaatttct acagcaactt ggcaagggta
     6001 attttgggag tgtggagatg tgccggtatg accctctaca ggacaacact ggggaggtgg
     6061 tcgctgtaaa aaagcttcag catagtactg aagagcacct aagagacttt gaaagggaaa
     6121 ttgaaatcct gaaatcccta cagcatgaca acattgtaaa gtacaaggga gtgtgctaca
     6181 gtgctggtc
//
