LOCUS       sequence-127245-        4446 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 4446)
  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 4446)
  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-PRKAA2
FEATURES             Location/Qualifiers
     source          1..4446
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(149..176)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(268..354)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     403..425
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     417..434
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     418..434
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    450..549
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2206..2305)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2322..2341)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2323..2341)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2346..2362)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2346..2362)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2793..3584
                     /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      3677..4265
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4166..4185
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4419..4436
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
      121 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
      181 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
      241 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
      301 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
      361 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
      421 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
      481 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
      541 aagttggcat ggctgagaag cagaagcacg acgggcgggt gaagatcgga cactacgtgc
      601 tgggcgacac gctgggcgtc ggcaccttcg gcaaagtgaa gattggagaa catcaattaa
      661 caggccataa agtggcagtt aaaatcttaa atagacagaa gattcgcagt ttagatgttg
      721 ttggaaaaat aaaacgagaa attcaaaatc taaaactctt tcgtcatcct catattatca
      781 aactatacca ggtgatcagc actccaacag atttttttat ggtaatggaa tatgtgtctg
      841 gaggtgaatt atttgactac atctgtaagc atggacgggt tgaagagatg gaagccaggc
      901 ggctctttca gcagattctg tctgctgtgg attactgtca taggcatatg gttgttcatc
      961 gagacctgaa accagagaat gtcctgttgg atgcacacat gaatgccaag atagccgatt
     1021 tcggattatc taatatgatg tcagatggtg aatttctgag aactagttgc ggatctccaa
     1081 attatgcagc acctgaagtc atctcaggca gattgtatgc aggtcctgaa gttgatatct
     1141 ggagctgtgg tgttatcttg tatgctcttc tttgtggcac cctcccattt gatgatgagc
     1201 atgtacctac gttatttaag aagatccgag ggggtgtctt ttatatccca gaatatctca
     1261 atcgttctgt cgccactctc ctgatgcata tgctgcaggt tgacccactg aaacgagcaa
     1321 ctatcaaaga cataagagag catgaatggt ttaaacaaga tttgcccagt tacttatttc
     1381 ctgaagaccc ttcctatgat gctaacgtca ttgatgatga ggctgtgaaa gaagtgtgtg
     1441 aaaaatttga atgtacagaa tcagaagtaa tgaacagttt atatagtggt gaccctcaag
     1501 accagcttgc agtggcttat catcttatca ttgacaatcg gagaataatg aaccaagcca
     1561 gtgagttcta cctcgcctct agtcctccat ctggttcttt tatggatgat agtgccatgc
     1621 atattccccc aggcctgaaa cctcatccag aaaggatgcc acctcttata gcagacagcc
     1681 ccaaagcaag atgtccattg gatgcactga atacgactaa gcccaaatct ttagctgtga
     1741 aaaaagccaa gtggcatctt ggaatccgaa gtcagagcaa accgtatgac attatggctg
     1801 aagtttaccg agctatgaag cagctggatt ttgaatggaa ggtagtgaat gcataccatc
     1861 ttcgtgtaag aagaaaaaat ccagtgactg gcaattacgt gaaaatgagc ttacaacttt
     1921 acctggttga taacaggagc tatcttttgg actttaaaag cattgatgat gaagtagtgg
     1981 agcagagatc tggttcctca acacctcagc gttcctgttc tgctgctggc ttacacagac
     2041 caagatcaag ttttgattcc acaactgcag agagccattc actttctggc tctctcactg
     2101 gctctttgac cggaagcaca ttgtcttcag tttcacctcg cctgggcagt cacaccatgg
     2161 atttttttga aatgtgtgcc agtctgatta ctactttagc ccgttgccca actttcttgt
     2221 acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta
     2281 tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt
     2341 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca
     2401 ttgcacaaga taaaaatata tcatcatgcc tcctctagac cagccaggac agaaatgcct
     2461 cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac
     2521 gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct
     2581 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt
     2641 tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc
     2701 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc
     2761 agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat
     2821 cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg
     2881 ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt
     2941 tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc
     3001 ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca
     3061 ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg
     3121 gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg
     3181 atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg
     3241 cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct
     3301 taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt
     3361 tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg
     3421 actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg
     3481 cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg
     3541 tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat
     3601 gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa
     3661 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3721 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3781 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     3841 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3901 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3961 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4021 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4081 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4141 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4201 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4261 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4321 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4381 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     4441 cggaag
//
