LOCUS       sequence-443280-        3507 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 3507)
  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 3507)
  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-AK2
FEATURES             Location/Qualifiers
     source          1..3507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      822..831
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1386..1485)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1502..1521)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1503..1521)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1526..1542)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1526..1542)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1973..2764
                     /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      2857..3445
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3346..3365
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agttggcatg gctcccagcg tgccagcggc agaacccgag tatcctaaag gcatccgggc
      721 cgtgctgctg gggcctcccg gggccggtaa agggacccag gcacccagat tggctgaaaa
      781 cttctgtgtc tgccatttag ctactgggga catgctgagg gccatggtgg cttctggctc
      841 agagctagga aaaaagctga aggcaactat ggatgctggg aaactggtga gtgatgaaat
      901 ggtagtggag ctcattgaga agaatttgga gacccccttg tgcaaaaatg gttttcttct
      961 ggatggcttc cctcggactg tgaggcaggc agaaatgctc gatgacctca tggagaagag
     1021 gaaagagaag cttgattctg tgattgaatt cagcatccca gactctctgc tgatccgaag
     1081 aatcacagga aggctgattc accccaagag tggccgttcc taccacgagg agttcaaccc
     1141 tccaaaagag cccatgaaag atgacatcac cggggaaccc ttgatccgtc gatcagatga
     1201 taatgaaaag gccttgaaaa tccgcctgca agcctaccac actcaaacca ccccactcat
     1261 agagtactac aggaaacggg ggatccactc cgccatcgat gcatcccaga cccccgatgt
     1321 cgtgttcgca agcatcctag cagccttctc caaagccaca tgtaaagact tggttatgtt
     1381 tatctaccca actttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     1441 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     1501 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     1561 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgcc tcctctagac
     1621 cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc
     1681 gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta
     1741 atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg
     1801 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat
     1861 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
     1921 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga
     1981 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca
     2041 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa
     2101 gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg
     2161 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct
     2221 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc
     2281 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga
     2341 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt
     2401 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt
     2461 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga
     2521 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc
     2581 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt
     2641 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc
     2701 agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa
     2761 ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     2821 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2881 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2941 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3001 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3061 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3121 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3181 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3241 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3301 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3361 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3421 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3481 cttttgctgg ccttttgctc acatgtt
//
