LOCUS       Exported                6816 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pDONR223-ERBB3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6816)
  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 Nov 24. ():.
  PUBMED    21107320
REFERENCE   2  (bases 1 to 6816)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6816
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      121..148
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      240..326
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     375..397
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     389..406
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     390..406
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    422..521
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4548..4647)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4664..4683)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4665..4683)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4688..4704)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4688..4704)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5135..5926
                     /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      6019..6607
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6508..6527
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6761..6778
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
       61 cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca
      121 aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg
      181 cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga
      241 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
      301 tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct
      361 agcatggatg ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc
      421 ccaaataatg attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgatgagca
      481 atgctttttt ataatgccaa ctttgtacaa aaaagttggc accatgaggg cgaacgacgc
      541 tctgcaggtg ctgggcttgc ttttcagcct ggcccggggc tccgaggtgg gcaactctca
      601 ggcagtgtgt cctgggactc tgaatggcct gagtgtgacc ggcgatgctg agaaccaata
      661 ccagacactg tacaagctct acgagaggtg tgaggtggtg atggggaacc ttgagattgt
      721 gctcacggga cacaatgccg acctctcctt cctgcagtgg attcgagaag tgacaggcta
      781 tgtcctcgtg gccatgaatg aattctctac tctaccattg cccaacctcc gcgtggtgcg
      841 agggacccag gtctacgatg ggaagtttgc catcttcgtc atgttgaact ataacaccaa
      901 ctccagccac gctctgcgcc agctccgctt gactcagctc accgagattc tgtcaggggg
      961 tgtttatatt gagaagaacg ataagctttg tcacatggac acaattgact ggagggacat
     1021 cgtgagggac cgagatgctg agatagtggt gaaggacaat ggcagaagct gtcccccctg
     1081 tcatgaggtt tgcaaggggc gatgctgggg tcctggatca gaagactgcc agacattgac
     1141 caagaccatc tgtgctcctc agtgtaatgg tcactgcttt gggcccaacc ccaaccagtg
     1201 ctgccatgat gagtgtgccg ggggctgctc aggccctcag gacacagact gctttgcctg
     1261 ccggcacttc aatgacagtg gagcctgtgt acctcgctgt ccacagcctc ttgtctacaa
     1321 caagctaact ttccagctgg aacccaatcc ccacaccaag tatcagtatg gaggagtttg
     1381 tgtagccagc tgtccccata actttgtggt ggatcaaaca tcctgtgtca gggcctgtcc
     1441 tcctgacaag atggaagtag ataaaaatgg gctcaagatg tgtgagcctt gtgggggact
     1501 atgtcccaaa gcctgtgagg gaacaggctc tgggagccgc ttccagactg tggactcgag
     1561 caacattgat ggatttgtga actgcaccaa gatcctgggc aacctggact ttctgatcac
     1621 cggcctcaat ggagacccct ggcacaagat ccctgccctg gacccagaga agctcaatgt
     1681 cttccggaca gtacgggaga tcacaggtta cctgaacatc cagtcctggc cgccccacat
     1741 gcacaacttc agtgtttttt ccaatttgac aaccattgga ggcagaagcc tctacaaccg
     1801 gggcttctca ttgttgatca tgaagaactt gaatgtcaca tctctgggct tccgatccct
     1861 gaaggaaatt agtgctgggc gtatctatat aagtgccaat aggcagctct gctaccacca
     1921 ctctttgaac tggaccaagg tgcttcgggg gcctacggaa gagcgactag acatcaagca
     1981 taatcggccg cgcagagact gcgtggcaga gggcaaagtg tgtgacccac tgtgctcctc
     2041 tgggggatgc tggggcccag gccctggtca gtgcttgtcc tgtcgaaatt atagccgagg
     2101 aggtgtctgt gtgacccact gcaactttct gaatggggag cctcgagaat ttgcccatga
     2161 ggccgaatgc ttctcctgcc acccggaatg ccaacccatg gagggcactg ccacatgcaa
     2221 tggctcgggc tctgatactt gtgctcaatg tgcccatttt cgagatgggc cccactgtgt
     2281 gagcagctgc ccccatggag tcctaggtgc caagggccca atctacaagt acccagatgt
     2341 tcagaatgaa tgtcggccct gccatgagaa ctgcacccag gggtgtaaag gaccagagct
     2401 tcaagactgt ttaggacaaa cactggtgct gatcggcaaa acccatctga caatggcttt
     2461 gacagtgata gcaggattgg tagtgatttt catgatgctg ggcggcactt ttctctactg
     2521 gcgtgggcgc cggattcaga ataaaagggc tatgaggcga tacttggaac ggggtgagag
     2581 catagagcct ctggacccca gtgagaaggc taacaaagtc ttggccagaa tcttcaaaga
     2641 gacagagcta aggaagctta aagtgcttgg ctcgggtgtc tttggaactg tgcacaaagg
     2701 agtgtggatc cctgagggtg aatcaatcaa gattccagtc tgcattaaag tcattgagga
     2761 caagagtgga cggcagagtt ttcaagctgt gacagatcat atgctggcca ttggcagcct
     2821 ggaccatgcc cacattgtaa ggctgctggg actatgccca gggtcatctc tgcagcttgt
     2881 cactcaatat ttgcctctgg gttctctgct ggatcatgtg agacaacacc ggggggcact
     2941 ggggccacag ctgctgctca actggggagt acaaattgcc aagggaatgt actaccttga
     3001 ggaacatggt atggtgcata gaaacctggc tgcccgaaac gtgctactca agtcacccag
     3061 tcaggttcag gtggcagatt ttggtgtggc tgacctgctg cctcctgatg ataagcagct
     3121 gctatacagt gaggccaaga ctccaattaa gtggatggcc cttgagagta tccactttgg
     3181 gaaatacaca caccagagtg atgtctggag ctatggtgtg acagtttggg agttgatgac
     3241 cttcggggca gagccctatg cagggctacg attggctgaa gtaccagacc tgctagagaa
     3301 gggggagcgg ttggcacagc cccagatctg cacaattgat gtctacatgg tgatggtcaa
     3361 gtgttggatg attgatgaga acattcgccc aacctttaaa gaactagcca atgagttcac
     3421 caggatggcc cgagacccac cacggtatct ggtcataaag agagagagtg ggcctggaat
     3481 agcccctggg ccagagcccc atggtctgac aaacaagaag ctagaggaag tagagctgga
     3541 gccagaacta gacctagacc tagacttgga agcagaggag gacaacctgg caaccaccac
     3601 actgggctcc gccctcagcc taccagttgg aacacttaat cggccacgtg ggagccagag
     3661 ccttttaagt ccatcatctg gatacatgcc catgaaccag ggtaatcttg gggagtcttg
     3721 ccaggagtct gcagtttctg ggagcagtga acggtgcccc cgtccagtct ctctacaccc
     3781 aatgccacgg ggatgcctgg catcagagtc atcagagggg catgtaacag gctctgaggc
     3841 tgagctccag gagaaagtgt caatgtgtag gagccggagc aggagccgga gcccacggcc
     3901 acgcggagat agcgcctacc attcccagcg ccacagtctg ctgactcctg ttaccccact
     3961 ctccccaccc gggttagagg aagaggatgt caacggttat gtcatgccag atacacacct
     4021 caaaggtact ccctcctccc gggaaggcac cctttcttca gtgggtctca gttctgtcct
     4081 gggtactgaa gaagaagatg aagatgagga gtatgaatac atgaaccgga ggagaaggca
     4141 cagtccacct catcccccta ggccaagttc ccttgaggag ctgggttatg agtacatgga
     4201 tgtggggtca gacctcagtg cctctctggg cagcacacag agttgcccac tccaccctgt
     4261 acccatcatg cccactgcag gcacaactcc agatgaagac tatgaatata tgaatcggca
     4321 acgagatgga ggtggtcctg ggggtgatta tgcagccatg ggggcctgcc cagcatctga
     4381 gcaagggtat gaagagatga gagcttttca ggggcctgga catcaggccc cccatgtcca
     4441 ttatgcccgc ctaaaaactc tacgtagctt agaggctaca gactctgcct ttgataaccc
     4501 tgattactgg catagcaggc ttttccccaa ggctaatgcc cagagaacgc caactttctt
     4561 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     4621 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     4681 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     4741 cattgcacaa gataaaaata tatcatcatg cctcctctag accagccagg acagaaatgc
     4801 ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc
     4861 acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg
     4921 ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt
     4981 tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca
     5041 gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca
     5101 gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga tcgccgaagt
     5161 atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct
     5221 ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga
     5281 tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga
     5341 ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac
     5401 cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt
     5461 tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat
     5521 tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc
     5581 ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac
     5641 cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac
     5701 gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc
     5761 cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca
     5821 ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt
     5881 tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct cgagccaccc
     5941 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     6001 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6061 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6121 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6181 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6241 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6301 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6361 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6421 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6481 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6541 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6601 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6661 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     6721 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     6781 agcggaagag cgcccaatac gcaaaccgcc tctccc
//
