LOCUS       Exported               11387 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 11387)
  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(pgk1 up and down)] as an 
            inserted sequence.
FEATURES             Location/Qualifiers
     source          1..11387)
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    complement(253..487))
                     /label=TEF TER
                     /note="color: #0000ff; direction: LEFT"
     misc_feature    complement(488..1516))
                     /label=HPH
                     /note="color: #ff7f00; direction: LEFT"
     misc_feature    complement(1517..1884))
                     /label=TEF PRO
                     /note="color: #400080; direction: LEFT"
                     1830..2920)
                     /note="note=""Fragment inserted from PCR(pgk1 up and down) 
                     into bRA66 bRA63 plus BplI gblock by homologous 
                     recombination"
                     /note="color: #84a4c0; direction: RIGHT"
     misc_feature    2878..7135)
                     /label=CaCas9
                     /note="color: #ff0000; direction: RIGHT"
     misc_feature    7039..7062)
                     /label=NLS
                     /note="color: #ff00ff; direction: RIGHT"
     CDS             7105..7128)
                     /label=Flag tag
                     /note="color: #008000; direction: RIGHT"
     misc_feature    7135..7403)
                     /label=CYC1 terminator
                     /note="note=""direct strand 
                     translation""translation=""SLLESCN*"""
                     /note="color: #0000ff; direction: RIGHT"
     rep_origin      complement(7454..7909))
                     /direction=LEFT
                     /label=M13 origin
                     /note="color: #4682b4"
     rep_origin      7464..7904)
                     /direction=RIGHT
                     /label=F1 ori
                     /note="color: #4682b4"
     CDS             complement(7914..7982))
                     /label=LacZ alpha
                     /note="color: #6495ed; direction: LEFT"
     primer_bind     8053..8070)
                     /label=M13-fwd
                     /note="color: #008b8b; direction: RIGHT"
     primer_bind     8077..8096)
                     /label=T7
                     /note="color: #008b8b; direction: RIGHT"
     misc_feature    8445..8471)
                     /label=BplI
                     /note="color: #ff0000; direction: RIGHT"
     primer_bind     complement(8611..8630))
                     /label=T3
                     /note="color: #008b8b; direction: LEFT"
     primer_bind     complement(8648..8668))
                     /label=M13-rev
                     /note="color: #008b8b; direction: LEFT"
     misc_binding    complement(8674..8696))
                     /label=LacO
                     /note="color: #6495ed; direction: LEFT"
     rep_origin      complement(9036..9664))
                     /direction=LEFT
                     /label=ColE1 origin
                     /note="color: #4682b4"
     CDS             complement(9816..10475))
                     /label=AmpR
                     /note="color: #8fbc8f; direction: LEFT"
     promoter        complement(10715..10743))
                     /label=Amp prom
                     /note="color: #4682b4; direction: LEFT"
     rep_origin      10805..11315)
                     /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 attcacatac gattgacgca tgatattact ttctgcgcac ttaacttcgc atctgggcag
      361 atgatgtcga ggcgaaaaaa aatataaatc acgctaacat ttgattaaaa tagaacaact
      421 acaatataaa aaaactatac aaatgacaag ttcttgaaaa caagaatctt tttattgtca
      481 gtactgatta ttcctttgcc ctcggacgag tgctggggcg tcggtttcca ctatcggcga
      541 gtacttctac acagccatcg gtccagacgg ccgcgcttct gcgggcgatt tgtgtacgcc
      601 cgacagtccc ggctccggat cggacgattg cgtcgcatcg accctgcgcc caagctgcat
      661 catcgaaatt gccgtcaacc aagctctgat agagttggtc aagaccaatg cggagcatat
      721 acgcccggag ccgcggcgat cctgcaagct ccggatgcct ccgctcgaag tagcgcgtct
      781 gctgctccat acaagccaac cacggcctcc agaagaagat gttggcgacc tcgtattggg
      841 aatccccgaa catcgcctcg ctccagtcaa tgaccgctgt tatgcggcca ttgtccgtca
      901 ggacattgtt ggagccgaaa tccgcgtgca cgaggtgccg gacttcgggg cagtcctcgg
      961 cccaaagcat cagctcatcg agagcctgcg cgacggacgc actgacggtg tcgtccatca
     1021 cagtttgcca gtgatacaca tggggatcag caatcgcgca tatgaaatca cgccatgtag
     1081 tgtattgacc gattccttgc ggtccgaatg ggccgaaccc gctcgtctgg ctaagatcgg
     1141 ccgcagcgat cgcatccatg gcctccgcga ccggctgcag aacagcgggc agttcggttt
     1201 caggcaggtc ttgcaacgtg acaccctgtg cacggcggga gatgcaatag gtcaggctct
     1261 cgctgaattc cccaatgtca agcacttccg gaatcgggag cgcggccgat gcaaagtgcc
     1321 gataaacata acgatctttg tagaaaccat cggcgcagct atttacccgc aggacatatc
     1381 cacgccctcc tacatcgaag ctgaaagcac gagattcttc gccctccgag agctgcatca
     1441 ggtcggagac gctgtcgaac ttttcgatca gaaacttctc gacagacgtc gcggtgagtt
     1501 caggcttttt acccatggtt gtttatgttc ggatgtgatg tgagaactgt atcctagcaa
     1561 gattttaaaa ggaagtatat gaaagaagaa cctcagtggc aaatcctaac cttttatatt
     1621 tctctacagg ggcgcggcgt ggggacaatt caacgcgtct gtgaggggag cgtttccctg
     1681 ctcgcaggtc tgcagcgagg agccgtaatt tttgcttcgc gccgtgcggc catcaaaatg
     1741 tatggatgca aatgattata catggggatg tatgggctaa atgtacgggc gacagtcaca
     1801 tcatgcccct gagctgcgca cgtcaagact gtcaaggagg gtattctggg cctccatgtc
     1861 gctggccggg tgacccggcg gggaaagtac cttcaaagaa tggggtctta tcttgttttg
     1921 caagtaccac tgagcaggat aataatagaa atgataatat actatagtag agataacgtc
     1981 gatgacttcc catactgtaa ttgcttttag ttgtgtattt ttagtgtgca agtttctgta
     2041 aatcgattaa tttttttttc tttcctcttt ttattaacct taatttttat tttagattcc
     2101 tgacttcaac tcaagacgca cagatattat aacatctgca taataggcat ttgcaagaat
     2161 tactcgtgag taaggaaaga gtgaggaact atcgcatacc tgcatttaaa gatgccgatt
     2221 tgggcgcgaa tcctttattt tggcttcacc ctcatactat tatcagggcc agaaaaagga
     2281 agtgtttccc tccttcttga attgatgtta ccctcataaa gcacgtggcc tcttatcgag
     2341 aaagaaatta ccgtcgctcg tgatttgttt gcaaaaagaa caaaactgaa aaaacccaga
     2401 cacgctcgac ttcctgtctt cctattgatt gcagcttcca atttcgtcac acaacaaggt
     2461 cctagcgacg gctcacaggt tttgtaacaa gcaatcgaag gttctggaat ggcgggaaag
     2521 ggtttagtac cacatgctat gatgcccact gtgatctcca gagcaaagtt cgttcgatcg
     2581 tactgttact ctctctcttt caaacagaat tgtccgaatc gtgtgacaac aacagcctgt
     2641 tctcacacac tcttttcttc taaccaaggg ggtggtttag tttagtagaa cctcgtgaaa
     2701 cttacattta catatatata aacttgcata aattggtcaa tgcaagaaat acatatttgg
     2761 tcttttctaa ttcgtagttt ttcaagttct tagatgcttt ctttttctct tttttacaga
     2821 tcatcaagga agtaattatc tactttttac aacaaatata aaacaccgga tctcaaaatg
     2881 gataaaaagt atagtattgg tttagatatt ggtactaact ctgtgggttg ggcagttatc
     2941 actgacgaat ataaagttcc atcaaagaaa tttaaggtgt taggtaacac tgacagacac
     3001 tcaataaaaa agaatcttat cggtgctctt ttgttcgact ccggtgaaac tgccgaggct
     3061 acacgtttaa aaagaacagc aagaagaaga tatacccgta gaaaaaatag aatatgttat
     3121 ttacaagaaa tcttttctaa tgaaatggct aaagttgatg attccttttt ccatagattg
     3181 gaagagtcat ttttggttga agaagacaaa aagcatgaga gacatccaat ctttgggaat
     3241 atagttgatg aagtggctta ccatgaaaaa tatcctacca tttatcattt aagaaagaaa
     3301 ttggtagatt caactgataa agctgacctt agattaatct atttagcact tgcccatatg
     3361 attaaattta gaggtcattt tttgattgaa ggtgatttga acccagataa ttctgacgtg
     3421 gataaattat ttattcaatt agtccaaacc tacaaccaat tatttgagga aaatccaatt
     3481 aatgctagtg gtgtcgatgc caaagctata ttatcagcca gattatcaaa atctagacgt
     3541 ttggaaaatt tgattgccca attgccagga gaaaaaaaga atggattatt tggaaacttg
     3601 atcgcattat cattgggttt gacaccaaat tttaaatcta attttgattt agctgaagat
     3661 gctaaattac aattatcaaa agacacctat gacgacgatt tggacaattt acttgctcaa
     3721 attggtgatc aatatgcaga tttgttctta gctgctaaaa acttatctga tgctattttg
     3781 ttgtctgata ttttgagagt gaacacagaa ataaccaaag ctccattatc agcatctatg
     3841 atcaaacgtt atgatgaaca ccatcaggat ttgactttat tgaaagcttt ggtgagacaa
     3901 caattgccag agaagtataa agaaatcttt ttcgatcaat ctaaaaacgg gtatgcaggt
     3961 tatattgatg ggggtgcctc ccaagaggaa ttttacaaat ttataaaacc tattttagaa
     4021 aagatggatg ggactgagga acttttggtc aaattgaaca gagaagattt gttacgtaaa
     4081 cagagaactt ttgataatgg tagtatacct caccaaattc atttgggtga gttgcatgca
     4141 attttaagaa gacaagaaga tttttatcca tttttaaaag ataatagaga aaaaatcgag
     4201 aaaattttaa cctttagaat tccatactat gttgggcctt tggctagagg taattcaaga
     4261 tttgcctgga tgacacgtaa atcagaagaa actattaccc cttggaattt tgaagaggtt
     4321 gttgataaag gagcatcagc acagagtttt attgaaagaa tgaccaattt cgataaaaac
     4381 ttaccaaatg aaaaagtttt accaaaacat tccttgttat acgaatattt tactgtttac
     4441 aatgaactta caaaggttaa atatgttact gaaggtatgc gtaagccagc ctttttatct
     4501 ggagaacaga aaaaggcaat agttgattta ttgtttaaaa caaatagaaa agttactgtt
     4561 aaacaattaa aagaagatta ctttaagaaa attgaatgtt ttgattcagt tgaaatcagt
     4621 ggtgttgaag acagatttaa tgctagttta ggaacttacc atgatttact taaaattatc
     4681 aaagataaag atttcttgga taacgaagaa aatgaagaca ttttagaaga cattgtttta
     4741 accttaactt tattcgaaga tagagagatg attgaagaac gtttgaagac ttatgcacat
     4801 ttgtttgacg ataaagtgat gaaacagttg aaaagaagac gttatactgg atggggtaga
     4861 ttgtctcgta aattgatcaa tggaattaga gataaacaaa gtggtaaaac tatcttggac
     4921 tttttgaaat ctgacggatt tgctaataga aatttcatgc aattgatcca cgacgatagt
     4981 ttgacattta aagaagacat ccaaaaggcc caagtgagtg ggcaaggtga ttcattacat
     5041 gaacatattg caaatttagc cggatctcct gctattaaga aagggatatt acaaactgtt
     5101 aaagttgtgg atgaattagt gaaagtaatg ggaagacata aacctgaaaa cattgtcatt
     5161 gagatggcaa gagaaaatca aactacacaa aaaggacaga aaaatagtag agaacgtatg
     5221 aaaagaatag aagagggtat taaagaattg ggtagtcaaa tattgaaaga acacccagtg
     5281 gaaaataccc agttgcaaaa tgaaaaatta tatctttact accttcaaaa tggacgtgat
     5341 atgtatgttg atcaggaatt agatataaat agactttcag attatgatgt agatcatata
     5401 gttccacaat ctttcttgaa agatgattcc atagacaata aagtattaac tagaagtgat
     5461 aaaaatagag gtaaaagtga taatgtccca agtgaggaag tcgtcaaaaa gatgaaaaat
     5521 tactggcgtc aacttttgaa tgctaaatta attactcaaa gaaaatttga taatttgact
     5581 aaagcagaaa gaggtgggct ttctgaatta gataaagccg ggttcattaa aagacaattg
     5641 gtcgaaacta gacaaattac taaacatgtt gcccaaattt tagattcccg tatgaacact
     5701 aagtatgacg aaaatgataa gttaatacgt gaggttaaag tcattacttt aaaatcaaaa
     5761 cttgtctctg atttcagaaa ggatttccaa ttctataaag ttagagaaat taataattat
     5821 catcatgctc atgatgcata tttgaatgct gtagttggaa ctgctttaat caagaaatac
     5881 cctaaattag aatctgaatt tgtatatggt gattacaaag tctatgatgt tagaaagatg
     5941 attgctaaat cagaacaaga aattggtaaa gctacagcta aatacttctt ttactctaac
     6001 attatgaatt tctttaaaac agaaattact ttggcaaacg gtgaaattag aaaaagacct
     6061 cttattgaaa caaatggtga gactggagag atagtttggg acaaagggcg tgatttcgct
     6121 actgttagaa aagttttatc aatgccacaa gttaacattg taaagaaaac agaggttcaa
     6181 actggtggtt tctcaaaaga aagtattttg cctaaaagaa atagtgataa attgattgcc
     6241 agaaaaaagg attgggatcc aaagaaatat ggtggtttcg actcaccaac cgtagcctat
     6301 tctgttttgg ttgtggcaaa ggttgaaaag ggtaaaagta aaaagcttaa atcagtaaaa
     6361 gaacttttgg gtattacaat aatggaaaga agttcctttg aaaagaaccc tattgatttt
     6421 ttggaagcta aaggttataa ggaagtaaag aaggacttaa taatcaaatt gcctaaatat
     6481 tctttatttg aattagaaaa tgggagaaaa agaatgttgg cttctgctgg agaattgcaa
     6541 aagggtaatg aattagcatt gccttccaaa tatgttaact tcttgtattt agcttcacac
     6601 tatgaaaagt tgaaagggtc accagaagat aacgagcaaa aacaattatt tgttgaacaa
     6661 cacaaacact acttagatga gattatagaa caaattagtg aattcagtaa aagagtgata
     6721 ttagctgatg caaatttaga taaagttttg tcagcctata acaaacatag agataagcca
     6781 attagagaac aagcagaaaa cattattcac ttatttaccc ttaccaattt aggagcacct
     6841 gctgctttca agtattttga tacaacaatt gatcgtaaaa gatatacctc aacaaaagaa
     6901 gtcttagacg ccaccttaat tcatcaatca atcactggat tgtatgagac aagaattgat
     6961 ttgtctcaat tgggtggtga tgaaggggct gatcctaaga agaaaagaaa agttgatcca
     7021 aagaaaaagc gtaaggtgga tcctaagaaa aagagaaagg ttgactacaa agaccatgac
     7081 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagtg ataatctctt
     7141 ctcgagtcat gtaattagtt atgtcacgct tacattcacg ccctcccccc acatccgctc
     7201 taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat ttttttatag
     7261 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacagac
     7321 gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga gaaggttttg ggacgctcga
     7381 aggctttaat ttgcggccgg tactgcggtg tgaaataccg cacagatgcg taaggagaaa
     7441 ataccgcatc aggaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt
     7501 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     7561 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     7621 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     7681 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     7741 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     7801 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     7861 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc gcgccattcg
     7921 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     7981 cagctggcga aggggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     8041 cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa
     8101 ttggagctcc accgcggtgg cggccgctct agaactagtg gatcccccgg gctgcaggaa
     8161 ttcgatatca agctttcttt gaaaagataa tgtatgatta tgctttcact catatttata
     8221 cagaaacttg atgttttctt tcgagtatat acaaggtgat tacatgtacg tttgaagtac
     8281 aactctagat tttgtagtgc cctcttgggc tagcggtaaa ggtgcgcatt ttttcacacc
     8341 ctacaatgtt ctgttcaaaa gattttggtc aaacgctgta gaagtgaaag ttggtgcgca
     8401 tgtttcggcg ttcgaaactt ctccgcagtg aaagataaat gatcctagat gcgagatatc
     8461 ctcatgccta ggttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa
     8521 cttgaaaaag tggcaccgag tcggtggtgc tttttttgtt ttttatgtct accgtcgacc
     8581 tcgagggggg gcccggtacc cagcttttgt tccctttagt gagggttaat tccgagcttg
     8641 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     8701 aacataggag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gaggtaactc
     8761 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     8821 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     8881 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     8941 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     9001 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     9061 aggctcggcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     9121 ccgacaggac tataaagata ccaggcgttc ccccctggaa gctccctcgt gcgctctcct
     9181 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     9241 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     9301 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     9361 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     9421 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     9481 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     9541 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     9601 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     9661 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     9721 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     9781 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     9841 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactgcccgt cgtgtagata
     9901 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     9961 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
    10021 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
    10081 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
    10141 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
    10201 gttacatgat cccccatgtt gtgaaaaaaa gcggttagct ccttcggtcc tccgatcgtt
    10261 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
    10321 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
    10381 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
    10441 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
    10501 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
    10561 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
    10621 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
    10681 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
    10741 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
    10801 cctgggtcct tttcatcacg tgctataaaa ataattataa tttaaatttt ttaatataaa
    10861 tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc
    10921 cgaagatgta aaagactcta gggggatcgc caacaaatac taccttttat cttgctcttc
    10981 ctgctctcag gtattaatgc cgaattgttt catcttgtct gtgtagaaga ccacacacga
    11041 aaatcctgtg attttacatt ttacttatcg ttaatcgaat gtatatctat ttaatctgct
    11101 tttcttgtct aataaatata tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt
    11161 gtttattttt ttttcttcat tccgtaactc ttctaccttc tttatttact ttctaaaatc
    11221 caaatacaaa acataaaaat aaataaacac agagtaaatt cccaaattat tccatcatta
    11281 aaagatacga ggcgcgtgta agttacaggc aagcgatccg tcctaagaaa ccattattat
    11341 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtc
//
