LOCUS       Exported               11398 bp ds-DNA     circular SYN 04-AUG-2017
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    SnapGene DNA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11398)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 2017 from SnapGene 4.0.1
            http://www.snapgene.com
COMMENT     Homologous recombination reaction between [bRA66 bRA63 plus BplI 
            gblock] as a target sequence and [PCR(LEU2 1kb up and down)] as an 
            inserted sequence.
FEATURES             Location/Qualifiers
     source          1..11398)
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        2424..2876)
                     /label=GAL1 promoter
                     /note="color: #00ff00; direction: RIGHT"
     misc_feature    2889..7146)
                     /label=CaCas9
                     /note="color: #ff0000; direction: RIGHT"
     misc_feature    7050..7073)
                     /label=NLS
                     /note="color: #ff00ff"
     CDS             7116..7139)
                     /label=Flag tag
                     /note="color: #008000; direction: RIGHT"
     misc_feature    7146..7414)
                     /label=CYC1 terminator
                     /note="note=""direct strand 
                     translation""translation=""SLLESCN*"""
                     /note="color: #0000ff"
     rep_origin      complement(7465..7920))
                     /direction=LEFT
                     /label=M13 origin
                     /note="color: #4682b4"
     rep_origin      7475..7915)
                     /direction=RIGHT
                     /label=F1 ori
                     /note="color: #4682b4"
     CDS             complement(7925..7993))
                     /label=LacZ alpha
                     /note="color: #6495ed; direction: LEFT"
     primer_bind     8064..8081)
                     /label=M13-fwd
                     /note="color: #008b8b; direction: RIGHT"
     primer_bind     8088..8107)
                     /label=T7
                     /note="color: #008b8b; direction: RIGHT"
     misc_feature    8187..8455)
                     /label=SNR52p
                     /note="color: #ffff00"
     misc_feature    8456..8482)
                     /label=BplI
                     /note="color: #ff0000"
     misc_feature    8483..8581)
                     /label=sup4 term + tracer
                     /note="color: #00ff00"
     primer_bind     complement(8622..8641))
                     /label=T3
                     /note="color: #008b8b; direction: LEFT"
     primer_bind     complement(8659..8679))
                     /label=M13-rev
                     /note="color: #008b8b; direction: LEFT"
     rep_origin      complement(9047..9675))
                     /direction=LEFT
                     /label=ColE1 origin
                     /note="color: #4682b4"
     CDS             complement(9827..10486))
                     /label=AmpR
                     /note="color: #8fbc8f; direction: LEFT"
     promoter        complement(10726..10754))
                     /label=Amp prom
                     /note="color: #4682b4; direction: LEFT"
     rep_origin      10816..11326)
                     /label=CEN-ARS pRS
                     /note="color: #ff6fcf"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accacgcttt tcaattcaat tcatcatttt ttttttattc ttttttttga tttcgaattc
      241 gagctcgttt aaactggatg gcggcgttag tatcgaatcg acagcagtat agcgaccagc
      301 attcacataa ggccgtttct gacagagtaa aattcttgag ggaactttca ccattatggg
      361 aaatggttca agaaggtatt gacttaaact ccatcaaatg gtcaggtcat tgagtgtttt
      421 ttatttgttg tatttttttt tttttagaga aaatcctcca atatataaat taggaatcat
      481 agtttcatga ttttctgtta cacctaactt tttgtgtggt gccctcctcc ttgtcaatat
      541 taatgttaaa gtgcaattct ttttccttat cacgttgagc cattagtatc aatttgctta
      601 cctgtattcc tttacatcct cctttttctc cttcttgata aatgtatgta gattgcgtat
      661 atagtttcgt ctaccctatg aacatattcc attttgtaat ttcgtgtcgt ttctattatg
      721 aatttcattt ataaagttta tgtacaaata tcataaaaaa agagaatctt tttaagcaag
      781 gattttctta acttcttcgg cgacagcatc accgacttcg gtggtactgt tggaaccacc
      841 taaatcacca gttctgatac ctgcatccaa aaccttttta actgcatctt caatggcctt
      901 accttcttca ggcaagttca atgacaattt caacatcatt gcagcagaca agatagtggc
      961 gatagggttg accttattct ttggcaaatc tggagcagaa ccgtggcatg gttcgtacaa
     1021 accaaatgcg gtgttcttgt ctggcaaaga ggccaaggac gcagatggca acaaacccaa
     1081 ggaacctggg ataacggagg cttcatcgga gatgatatca ccaaacatgt tgctggtgat
     1141 tataatacca tttaggtggg ttgggttctt aactaggatc atggcggcag aatcaatcaa
     1201 ttgatgttga accttcaatg tagggaattc gttcttgatg gtttcctcca cagtttttct
     1261 ccataatctt gaagaggcca aaacattagc tttatccaag gaccaaatag gcaatggtgg
     1321 ctcatgttgt agggccatga aagcggccat tcttgtgatt ctttgcactt ctggaacggt
     1381 gtattgttca ctatcccaag cgacaccatc accatcgtct tcctttctct taccaaagta
     1441 aatacctccc actaattctc tgacaacaac gaagtcagta cctttagcaa attgtggctt
     1501 gattggagat aagtctaaaa gagagtcgga tgcaaagtta catggtctta agttggcgta
     1561 caattgaagt tctttacgga tttttagtaa accttgttca ggtctaacac taccggtacc
     1621 ccatttagga ccacccacag cacctaacaa aacggcatca gccttcttgg aggcttccag
     1681 cgcctcatct ggaagtggaa cacctgtagc atcgatagca gcaccaccaa ttaaatgatt
     1741 ttcgaaatcg aacttgacat tggaacgaac atcagaaata gctttaagaa ccttaatggc
     1801 ttcggctgtg atttcttgac caacgtggtc acctggcaaa acgacgatct tcttaggggc
     1861 agacattaga atggtatatc cttgaaatat atatatatat attgctgaaa tgtaaaaggt
     1921 aagaaaagtt agaaagtaag acgattgcta accacctatt ggaaaaaaca ataggtcctt
     1981 aaataatatt gtcaacttca agtattgtga tgcaagcatt tagtcatgaa cgcttctcta
     2041 ttctatatga aaagccggtt ccggcgctct cacctttcct ttttctccca atttttcagt
     2101 tgaaaaaggt atatgcgtca ggcgacctct gaaattaaca aaaaatttcc agtcatcgaa
     2161 tttgattctg tgcgatagcg cccctgtgtg ttctcgttat gttgaggaaa aaaataatgg
     2221 ttgctaagag attcgaactc ttgcatctta cgatacctga gtattcccac agttgtattc
     2281 tgggcctcca tgtcgctggc cgggtgaccc ggcggggacg aggcaagcta aacagatctg
     2341 taaagagccc cattatctta gcctaaaaaa accttctctt tggaactttc agtaatacgc
     2401 ttaactgctc attgctatat tgaagtacgg attagaagcc gccgagcggg tgacagccct
     2461 ccgaaggaag actctcctcc gtgcgtcctc gtcttcaccg gtcgcgttcc tgaaacgcag
     2521 atgtgcctcg cgccgcactg ctccgaacaa taaagattct acaatactag cttttatggt
     2581 tatgaagagg aaaaattggc agtaacctgg ccccacaaac cttcaaatga acgaatcaaa
     2641 ttaacaacca taggatgata atgcgattag ttttttagcc ttatttctgg ggtaattaat
     2701 cagcgaagcg atgatttttg atctattaac agatatataa atgcaaaaac tgcataacca
     2761 ctttaactaa tactttcaac attttcggtt tgtattactt cttattcaaa tgtaataaaa
     2821 gtatcaacaa aaaattgtta atatacctct atactttaac gtcaaggaga aaaaacccgg
     2881 atctcaaaat ggataaaaag tatagtattg gtttagatat tggtactaac tctgtgggtt
     2941 gggcagttat cactgacgaa tataaagttc catcaaagaa atttaaggtg ttaggtaaca
     3001 ctgacagaca ctcaataaaa aagaatctta tcggtgctct tttgttcgac tccggtgaaa
     3061 ctgccgaggc tacacgttta aaaagaacag caagaagaag atatacccgt agaaaaaata
     3121 gaatatgtta tttacaagaa atcttttcta atgaaatggc taaagttgat gattcctttt
     3181 tccatagatt ggaagagtca tttttggttg aagaagacaa aaagcatgag agacatccaa
     3241 tctttgggaa tatagttgat gaagtggctt accatgaaaa atatcctacc atttatcatt
     3301 taagaaagaa attggtagat tcaactgata aagctgacct tagattaatc tatttagcac
     3361 ttgcccatat gattaaattt agaggtcatt ttttgattga aggtgatttg aacccagata
     3421 attctgacgt ggataaatta tttattcaat tagtccaaac ctacaaccaa ttatttgagg
     3481 aaaatccaat taatgctagt ggtgtcgatg ccaaagctat attatcagcc agattatcaa
     3541 aatctagacg tttggaaaat ttgattgccc aattgccagg agaaaaaaag aatggattat
     3601 ttggaaactt gatcgcatta tcattgggtt tgacaccaaa ttttaaatct aattttgatt
     3661 tagctgaaga tgctaaatta caattatcaa aagacaccta tgacgacgat ttggacaatt
     3721 tacttgctca aattggtgat caatatgcag atttgttctt agctgctaaa aacttatctg
     3781 atgctatttt gttgtctgat attttgagag tgaacacaga aataaccaaa gctccattat
     3841 cagcatctat gatcaaacgt tatgatgaac accatcagga tttgacttta ttgaaagctt
     3901 tggtgagaca acaattgcca gagaagtata aagaaatctt tttcgatcaa tctaaaaacg
     3961 ggtatgcagg ttatattgat gggggtgcct cccaagagga attttacaaa tttataaaac
     4021 ctattttaga aaagatggat gggactgagg aacttttggt caaattgaac agagaagatt
     4081 tgttacgtaa acagagaact tttgataatg gtagtatacc tcaccaaatt catttgggtg
     4141 agttgcatgc aattttaaga agacaagaag atttttatcc atttttaaaa gataatagag
     4201 aaaaaatcga gaaaatttta acctttagaa ttccatacta tgttgggcct ttggctagag
     4261 gtaattcaag atttgcctgg atgacacgta aatcagaaga aactattacc ccttggaatt
     4321 ttgaagaggt tgttgataaa ggagcatcag cacagagttt tattgaaaga atgaccaatt
     4381 tcgataaaaa cttaccaaat gaaaaagttt taccaaaaca ttccttgtta tacgaatatt
     4441 ttactgttta caatgaactt acaaaggtta aatatgttac tgaaggtatg cgtaagccag
     4501 cctttttatc tggagaacag aaaaaggcaa tagttgattt attgtttaaa acaaatagaa
     4561 aagttactgt taaacaatta aaagaagatt actttaagaa aattgaatgt tttgattcag
     4621 ttgaaatcag tggtgttgaa gacagattta atgctagttt aggaacttac catgatttac
     4681 ttaaaattat caaagataaa gatttcttgg ataacgaaga aaatgaagac attttagaag
     4741 acattgtttt aaccttaact ttattcgaag atagagagat gattgaagaa cgtttgaaga
     4801 cttatgcaca tttgtttgac gataaagtga tgaaacagtt gaaaagaaga cgttatactg
     4861 gatggggtag attgtctcgt aaattgatca atggaattag agataaacaa agtggtaaaa
     4921 ctatcttgga ctttttgaaa tctgacggat ttgctaatag aaatttcatg caattgatcc
     4981 acgacgatag tttgacattt aaagaagaca tccaaaaggc ccaagtgagt gggcaaggtg
     5041 attcattaca tgaacatatt gcaaatttag ccggatctcc tgctattaag aaagggatat
     5101 tacaaactgt taaagttgtg gatgaattag tgaaagtaat gggaagacat aaacctgaaa
     5161 acattgtcat tgagatggca agagaaaatc aaactacaca aaaaggacag aaaaatagta
     5221 gagaacgtat gaaaagaata gaagagggta ttaaagaatt gggtagtcaa atattgaaag
     5281 aacacccagt ggaaaatacc cagttgcaaa atgaaaaatt atatctttac taccttcaaa
     5341 atggacgtga tatgtatgtt gatcaggaat tagatataaa tagactttca gattatgatg
     5401 tagatcatat agttccacaa tctttcttga aagatgattc catagacaat aaagtattaa
     5461 ctagaagtga taaaaataga ggtaaaagtg ataatgtccc aagtgaggaa gtcgtcaaaa
     5521 agatgaaaaa ttactggcgt caacttttga atgctaaatt aattactcaa agaaaatttg
     5581 ataatttgac taaagcagaa agaggtgggc tttctgaatt agataaagcc gggttcatta
     5641 aaagacaatt ggtcgaaact agacaaatta ctaaacatgt tgcccaaatt ttagattccc
     5701 gtatgaacac taagtatgac gaaaatgata agttaatacg tgaggttaaa gtcattactt
     5761 taaaatcaaa acttgtctct gatttcagaa aggatttcca attctataaa gttagagaaa
     5821 ttaataatta tcatcatgct catgatgcat atttgaatgc tgtagttgga actgctttaa
     5881 tcaagaaata ccctaaatta gaatctgaat ttgtatatgg tgattacaaa gtctatgatg
     5941 ttagaaagat gattgctaaa tcagaacaag aaattggtaa agctacagct aaatacttct
     6001 tttactctaa cattatgaat ttctttaaaa cagaaattac tttggcaaac ggtgaaatta
     6061 gaaaaagacc tcttattgaa acaaatggtg agactggaga gatagtttgg gacaaagggc
     6121 gtgatttcgc tactgttaga aaagttttat caatgccaca agttaacatt gtaaagaaaa
     6181 cagaggttca aactggtggt ttctcaaaag aaagtatttt gcctaaaaga aatagtgata
     6241 aattgattgc cagaaaaaag gattgggatc caaagaaata tggtggtttc gactcaccaa
     6301 ccgtagccta ttctgttttg gttgtggcaa aggttgaaaa gggtaaaagt aaaaagctta
     6361 aatcagtaaa agaacttttg ggtattacaa taatggaaag aagttccttt gaaaagaacc
     6421 ctattgattt tttggaagct aaaggttata aggaagtaaa gaaggactta ataatcaaat
     6481 tgcctaaata ttctttattt gaattagaaa atgggagaaa aagaatgttg gcttctgctg
     6541 gagaattgca aaagggtaat gaattagcat tgccttccaa atatgttaac ttcttgtatt
     6601 tagcttcaca ctatgaaaag ttgaaagggt caccagaaga taacgagcaa aaacaattat
     6661 ttgttgaaca acacaaacac tacttagatg agattataga acaaattagt gaattcagta
     6721 aaagagtgat attagctgat gcaaatttag ataaagtttt gtcagcctat aacaaacata
     6781 gagataagcc aattagagaa caagcagaaa acattattca cttatttacc cttaccaatt
     6841 taggagcacc tgctgctttc aagtattttg atacaacaat tgatcgtaaa agatatacct
     6901 caacaaaaga agtcttagac gccaccttaa ttcatcaatc aatcactgga ttgtatgaga
     6961 caagaattga tttgtctcaa ttgggtggtg atgaaggggc tgatcctaag aagaaaagaa
     7021 aagttgatcc aaagaaaaag cgtaaggtgg atcctaagaa aaagagaaag gttgactaca
     7081 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     7141 gataatctct tctcgagtca tgtaattagt tatgtcacgc ttacattcac gccctccccc
     7201 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta
     7261 tttttttata gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt
     7321 tctgtacaga cgcgtgtacg catgtaacat tatactgaaa accttgcttg agaaggtttt
     7381 gggacgctcg aaggctttaa tttgcggccg gtactgcggt gtgaaatacc gcacagatgc
     7441 gtaaggagaa aataccgcat caggaaattg taaacgttaa tattttgtta aaattcgcgt
     7501 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     7561 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     7621 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     7681 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     7741 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     7801 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     7861 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     7921 cgcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     7981 cgctattacg ccagctggcg aaggggggat gtgctgcaag gcgattaagt tgggtaacgc
     8041 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa tacgactcac
     8101 tatagggcga attggagctc caccgcggtg gcggccgctc tagaactagt ggatcccccg
     8161 ggctgcagga attcgatatc aagctttctt tgaaaagata atgtatgatt atgctttcac
     8221 tcatatttat acagaaactt gatgttttct ttcgagtata tacaaggtga ttacatgtac
     8281 gtttgaagta caactctaga ttttgtagtg ccctcttggg ctagcggtaa aggtgcgcat
     8341 tttttcacac cctacaatgt tctgttcaaa agattttggt caaacgctgt agaagtgaaa
     8401 gttggtgcgc atgtttcggc gttcgaaact tctccgcagt gaaagataaa tgatcctaga
     8461 tgcgagatat cctcatgcct aggttttaga gctagaaata gcaagttaaa ataaggctag
     8521 tccgttatca acttgaaaaa gtggcaccga gtcggtggtg ctttttttgt tttttatgtc
     8581 taccgtcgac ctcgaggggg ggcccggtac ccagcttttg ttccctttag tgagggttaa
     8641 ttccgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     8701 caattccaca caacatagga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     8761 tgaggtaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     8821 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     8881 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     8941 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     9001 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     9061 cgtttttcca taggctcggc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     9121 ggtggcgaaa cccgacagga ctataaagat accaggcgtt cccccctgga agctccctcg
     9181 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     9241 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     9301 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     9361 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     9421 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     9481 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     9541 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     9601 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     9661 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     9721 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     9781 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     9841 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactgcccg
     9901 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     9961 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
    10021 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
    10081 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
    10141 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
    10201 gatcaaggcg agttacatga tcccccatgt tgtgaaaaaa agcggttagc tccttcggtc
    10261 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
    10321 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
    10381 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
    10441 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
    10501 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
    10561 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
    10621 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
    10681 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
    10741 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
    10801 gaaaagtgcc acctgggtcc ttttcatcac gtgctataaa aataattata atttaaattt
    10861 tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag
    10921 tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta
    10981 tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag
    11041 accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta
    11101 tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt
    11161 tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac
    11221 tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta
    11281 ttccatcatt aaaagatacg aggcgcgtgt aagttacagg caagcgatcc gtcctaagaa
    11341 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
