LOCUS       sequence-331347-        5787 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 5787)
  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 5787)
  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-MERTK
FEATURES             Location/Qualifiers
     source          1..5787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3130..3229)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3246..3265)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3247..3265)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3270..3286)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3270..3286)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3717..4508
                     /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      4601..5189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5090..5109
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5343..5360
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5519..5546)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5638..5724)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(5773..5787,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(5787,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagttgg caccatgggg ccggccccgc tgccgctgct gctgggcctc ttcctccccg
      181 cgctctggcg tagagctatc actgaggcaa gggaagaagc caagccttac ccgctattcc
      241 cgggaccttt tccagggagc ctgcaaactg accacacacc gctgttatcc cttcctcacg
      301 ccagtgggta ccagcctgcc ttgatgtttt caccaaccca gcctggaaga ccacatacag
      361 gaaacgtagc cattccccag gtgacctctg tcgaatcaaa gcccctaccg cctcttgcct
      421 tcaaacacac agttggacac ataatacttt ctgaacataa aggtgtcaaa tttaattgct
      481 caatcagtgt acctaatata taccaggaca ccacaatttc ttggtggaaa gatgggaagg
      541 aattgcttgg ggcacatcat gcaattacac agttttatcc agatgatgaa gttacagcaa
      601 taatcgcttc cttcagcata accagtgtgc agcgttcaga caatgggtcg tatatctgta
      661 agatgaaaat aaacaatgaa gagatcgtgt ctgatcccat ctacatcgaa gtacaaggac
      721 ttcctcactt tactaagcag cctgagagca tgaatgtcac cagaaacaca gccttcaacc
      781 tcacctgtca ggctgtgggc ccgcctgagc ccgtcaacat tttctgggtt caaaacagta
      841 gccgtgttaa cgaacagcct gaaaaatccc cctccgtgct aactgttcca ggcctgacgg
      901 agatggcggt cttcagttgt gaggcccaca atgacaaagg gctgaccgtg tccaagggag
      961 tgcagatcaa catcaaagca attccctccc caccaactga agtcagcatc cgtaacagca
     1021 ctgcacacag cattctgatc tcctgggttc ctggttttga tggatactcc ccgttcagga
     1081 attgcagcat tcaggtcaag gaagctgatc cgctgagtaa tggctcagtc atgattttta
     1141 acacctctgc cttaccacat ctgtaccaaa tcaagcagct gcaagccctg gctaattaca
     1201 gcattggtgt ttcctgcatg aatgaaatag gctggtctgc agtgagccct tggattctag
     1261 ccagcacgac tgaaggagcc ccatcagtag cacctttaaa tgtcactgtg tttctgaatg
     1321 aatctagtga taatgtggac atcagatgga tgaagcctcc gactaagcag caggatggag
     1381 aactggtggg ctaccggata tcccacgtgt ggcagagtgc agggatttcc aaagagctct
     1441 tggaggaagt tggccagaat ggcagccgag ctcggatctc tgttcaagtc cacaatgcta
     1501 cgtgcacagt gaggattgca gccgtcacca aagggggagt tgggcccttc agtgatccag
     1561 tgaaaatatt tatccctgca cacggttggg tagattatgc cccctcttca actccggcgc
     1621 ctggcaatgc agatcctgtg ctcatcatct ttggctgctt ttgtggattt attttgattg
     1681 ggttggtttt atacatctcc ttggccatca gaaaaagagt ccaggagaca aagtttggga
     1741 atgcattcac agaggaggat tctgaattag tggtgaatta tatagcaaag aaatccttct
     1801 gtcggcgagc cattgaactt accttacata gcttgggagt cagtgaggaa ctacaaaata
     1861 aactagaaga tgttgtgatt gacaggaatc ttctaattct tggaaaaatt ctgggtgaag
     1921 gagagtttgg gtctgtaatg gaaggaaatc ttaagcagga agatgggacc tctctgaaag
     1981 tggcagtgaa gaccatgaag ttggacaact cttcgcagcg ggagatcgag gagtttctca
     2041 gtgaggcagc gtgcatgaaa gacttcagcc acccaaatgt cattcgactt ctaggtgtgt
     2101 gtatagaaat gagctctcaa ggcatcccaa agcccatggt aattttaccc ttcatgaaat
     2161 acggggacct gcatacttac ttactttatt cccgattgga gacaggacca aagcatattc
     2221 ctctgcagac actattgaag ttcatggtgg atattgccct gggaatggag tatctgagca
     2281 acaggaattt tcttcatcga gatttagctg ctcgaaactg catgttgcga gatgacatga
     2341 ctgtctgtgt tgcggacttc ggcctctcta agaagattta cagtggcgat tattaccgcc
     2401 aaggccgcat tgctaagatg cctgttaaat ggatcgccat agaaagtctt gcagaccgag
     2461 tctacacaag taaaagtgat gtgtgggcat ttggcgtgac catgtgggaa atagctacgc
     2521 ggggaatgac tccctatcct ggggtccaga accatgagat gtatgactat cttctccatg
     2581 gccacaggtt gaagcagccc gaagactgcc tggatgaact gtatgaaata atgtactctt
     2641 gctggagaac cgatccctta gaccgcccca ccttttcagt attgaggctg cagctagaaa
     2701 aactcttaga aagtttgcct gacgttcgga accaagcaga cgttatttac gtcaatacac
     2761 agttgctgga gagctctgag ggcctggccc agggctccac ccttgctcca ctggacttga
     2821 acatcgaccc tgactctata attgcctcct gcactccccg cgctgccatc agtgtggtca
     2881 cagcagaagt tcatgacagc aaacctcatg aaggacggta catcctgaat gggggcagtg
     2941 aggaatggga agatctgact tctgccccct ctgctgcagt cacagctgaa aagaacagtg
     3001 ttttaccggg ggagagactt gttaggaatg gggtctcctg gtcccattcg agcatgctgc
     3061 ccttgggaag ctcattgccc gatgaacttt tgtttgctga cgactcctca gaaggctcag
     3121 aagtcctgat gccaactttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa
     3181 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
     3241 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
     3301 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgcctcctct
     3361 agaccagcca ggacagaaat gcctcgactt cgctgctacc caaggttgcc gggtgacgca
     3421 caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg gttcgtaagc
     3481 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
     3541 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt ggggtacagt
     3601 ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat
     3661 ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta aacattatga
     3721 gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc
     3781 gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc
     3841 tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa
     3901 cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga
     3961 ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc
     4021 cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct
     4081 tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata
     4141 gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc
     4201 tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg
     4261 atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa
     4321 tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc
     4381 ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc
     4441 gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg
     4501 gcaaataacc ctcgagccac ccatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
     4561 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4621 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4681 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4741 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4801 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4861 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4921 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4981 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5041 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5101 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5161 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5221 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     5281 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     5341 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     5401 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     5461 gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa
     5521 aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg
     5581 tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt
     5641 tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc
     5701 cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg ttaacgctag
     5761 catggatgtt ttcccagtca cgacgtt
//
