LOCUS       sequence-230411-        6804 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 6804)
  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 6804)
  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-FLT1
FEATURES             Location/Qualifiers
     source          1..6804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(154..181)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(273..359)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     408..430
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     422..439
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     423..439
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    455..554
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4569..4668)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4685..4704)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4686..4704)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4709..4725)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4709..4725)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5156..5947
                     /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      6040..6628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6529..6548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6782..6799
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
       61 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg
      121 taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
      181 tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
      241 tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
      301 gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga
      361 tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg
      421 ttgtaaaacg acggccagtc ttaagctcgg gccccaaata atgattttat tttgactgat
      481 agtgacctgt tcgttgcaac aaattgatga gcaatgcttt tttataatgc caactttgta
      541 caaaaaagtt ggcaccatgg tcagctactg ggacaccggg gtcctgctgt gcgcgctgct
      601 cagctgtctg cttctcacag gatctagttc aggttcaaaa ttaaaagatc ctgaactgag
      661 tttaaaaggc acccagcaca tcatgcaagc aggccagaca ctgcatctcc aatgcagggg
      721 ggaagcagcc cataaatggt ctttgcctga aatggtgagt aaggaaagcg aaaggctgag
      781 cataactaaa tctgcctgtg gaagaaatgg caaacaattc tgcagtactt taaccttgaa
      841 cacagctcaa gcaaaccaca ctggcttcta cagctgcaaa tatctagctg tacctacttc
      901 aaagaagaag gaaacagaat ctgcaatcta tatatttatt agtgatacag gtagaccttt
      961 cgtagagatg tacagtgaaa tccccgaaat tatacacatg actgaaggaa gggagctcgt
     1021 cattccctgc cgggttacgt cacctaacat cactgttact ttaaaaaagt ttccacttga
     1081 cactttgatc cctgatggaa aacgcataat ctgggacagt agaaagggct tcatcatatc
     1141 aaatgcaacg tacaaagaaa tagggcttct gacctgtgaa gcaacagtca atgggcattt
     1201 gtataagaca aactatctca cacatcgaca aaccaataca atcatagatg tccaaataag
     1261 cacaccacgc ccagtcaaat tacttagagg ccatactctt gttctcaatt gtactgctac
     1321 cactcccttg aacacgagag ttcaaatgac ctggagttac cctgatgaaa aaaataagag
     1381 agcttccgta aggcgacgaa ttgaccaaag caattcccat gccaacatat tctacagtgt
     1441 tcttactatt gacaaaatgc agaacaaaga caaaggactt tatacttgtc gtgtaaggag
     1501 tggaccatca ttcaaatctg ttaacacctc agtgcatata tatgataaag cattcatcac
     1561 tgtgaaacat cgaaaacagc aggtgcttga aaccgtagct ggcaagcggt cttaccggct
     1621 ctctatgaaa gtgaaggcat ttccctcgcc ggaagttgta tggttaaaag atgggttacc
     1681 tgcgactgag aaatctgctc gctatttgac tcgtggctac tcgttaatta tcaaggacgt
     1741 aactgaagag gatgcaggga attatacaat cttgctgagc ataaaacagt caaatgtgtt
     1801 taaaaacctc actgccactc taattgtcaa tgtgaaaccc cagatttacg aaaaggctgt
     1861 gtcatcgttt ccagacccgg ctctctaccc actgggcagc agacaaatcc tgacttgtac
     1921 cgcatatggt atccctcaac ctacaatcaa gtggttctgg cacccctgta accataatca
     1981 ttccgaagca aggtgtgact tttgttccaa taatgaagag tcctttatcc tggatgctga
     2041 cagcaacatg ggaaacagaa ttgagagcat cactcagcgc atggcaataa tagaaggaaa
     2101 gaataagatg gctagcacct tggttgtggc tgactctaga atttctggaa tctacatttg
     2161 catagcttcc aataaagttg ggactgtggg aagaaacata agcttttata tcacagatgt
     2221 gccaaatggg tttcatgtta acttggaaaa aatgccgacg gaaggagagg acctgaaact
     2281 gtcttgcaca gttaacaagt tcttatacag agacgttact tggattttac tgcggacagt
     2341 taataacaga acaatgcact acagtattag caagcaaaaa atggccatca ctaaggagca
     2401 ctccatcact cttaatctta ccatcatgaa tgtttccctg caagattcag gcacctatgc
     2461 ctgcagagcc aggaatgtat acacagggga agaaatcctc cagaagaaag aaattacaat
     2521 cagagatcag gaagcaccat acctcctgcg aaacctcagt gatcacacag tggccatcag
     2581 cagttccacc actttagact gtcatgctaa tggtgtcccc gagcctcaga tcacttggtt
     2641 taaaaacaac cacaaaatac aacaagagcc tggaattatt ttaggaccag gaagcagcac
     2701 gctgtttatt gaaagagtca cagaagagga tgaaggtgtc tatcactgca aagccaccaa
     2761 ccagaagggc tctgtggaaa gttcagcata cctcactgtt caaggaacct cggacaagcc
     2821 taatctggag ctgatcactc taacatgcac ctgtgtggct gcgactctct tctggctcct
     2881 attaaccctc tttatccgaa aaatgaaaag gtcttcttct gaaataaaga ctgactacct
     2941 atcaattata atggacccag atgaagttcc tttggatgag cagtgtgagc ggctccctta
     3001 tgatgccagc aagtgggagt ttgcccggga gagacttaaa ctgggcaaat cacttggaag
     3061 aggggctttt ggaaaagtgg ttcaagcatc agcatttggc attaagaaat cacctacgtg
     3121 ccggactgtg gctgtgaaaa tgctgaaaga gggggccacg gccagcgagt acaaagctct
     3181 gatgactgag ctaaaaatct tgacccacat tggccaccat ctgaacgtgg ttaacctgct
     3241 gggagcctgc accaagcaag gagggcctct gatggtgatt gttgaatact gcaaatatgg
     3301 aaatctctcc aactacctca agagcaaacg tgacttattt tttctcaaca aggatgcagc
     3361 actacacatg gagcctaaga aagaaaaaat ggagccaggc ctggaacaag gcaagaaacc
     3421 aagactagat agcgtcacca gcagcgaaag ctttgcgagc tccggctttc aggaagataa
     3481 aagtctgagt gatgttgagg aagaggagga ttctgacggt ttctacaagg agcccatcac
     3541 tatggaagat ctgatttctt acagttttca agtggccaga ggcatggagt tcctgtcttc
     3601 cagaaagtgc attcatcggg acctggcagc gagaaacatt cttttatctg agaacaacgt
     3661 ggtgaagatt tgtgattttg gccttgcccg ggatatttat aagaaccccg attatgtgag
     3721 aaaaggagat actcgacttc ctctgaaatg gatggctcct gaatctatct ttgacaaaat
     3781 ctacagcacc aagagcgacg tgtggtctta cggagtattg ctgtgggaaa tcttctcctt
     3841 aggtgggtct ccatacccag gagtacaaat ggatgaggac ttttgcagtc gcctgaggga
     3901 aggcatgagg atgagagctc ctgagtactc tactcctgaa atctatcaga tcatgctgga
     3961 ctgctggcac agagacccaa aagaaaggcc aagatttgca gaacttgtgg aaaaactagg
     4021 tgatttgctt caagcaaatg tacaacagga tggtaaagac tacatcccaa tcaatgccat
     4081 actgacagga aatagtgggt ttacatactc aactcctgcc ttctctgagg acttcttcaa
     4141 ggaaagtatt tcagctccga agtttaattc aggaagctct gatgatgtca gatacgtaaa
     4201 tgctttcaag ttcatgagcc tggaaagaat caaaaccttt gaagaacttt taccgaatgc
     4261 cacctccatg tttgatgact accagggcga cagcagcact ctgttggcct ctcccatgct
     4321 gaagcgcttc acctggactg acagcaaacc caaggcctcg ctcaagattg acttgagagt
     4381 aaccagtaaa agtaaggagt cggggctgtc tgatgtcagc aggcccagtt tctgccattc
     4441 cagctgtggg cacgtcagcg aaggcaagcg caggttcacc tacgaccacg ctgagctgga
     4501 aaggaaaatc gcgtgctgct ccccgccccc agactacaac tcggtggtcc tgtactccac
     4561 cccacccatc ccaactttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
     4621 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctga
     4681 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
     4741 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gcctcctcta
     4801 gaccagccag gacagaaatg cctcgacttc gctgctaccc aaggttgccg ggtgacgcac
     4861 accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct
     4921 gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg
     4981 tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc
     5041 tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg
     5101 gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa acattatgag
     5161 ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg
     5221 ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct
     5281 gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac
     5341 gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat
     5401 tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc
     5461 agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt
     5521 cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag
     5581 cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct
     5641 atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga
     5701 tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat
     5761 cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc
     5821 cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg
     5881 cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg
     5941 caaataaccc tcgagccacc catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     6001 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6061 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6121 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6181 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6241 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6301 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6361 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6421 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6481 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6541 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6601 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6661 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6721 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6781 cagcgagtca gtgagcgagg aagc
//
