LOCUS       Exported               10964 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 10964)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 2017 from SnapGene 4.0.1
            http://www.snapgene.com
COMMENT     Homologous recombination reaction between [bRA63 CaCas9 in bRA48] as
            a target sequence and [Sequence Window #4] as an inserted sequence.
FEATURES             Location/Qualifiers
     source          1..10964)
                     /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"
     promoter        1990..2442)
                     /label=GAL1 promoter
                     /note="color: #00ff00; direction: RIGHT"
     misc_feature    2455..6712)
                     /label=CaCas9
                     /note="color: #ff0000; direction: RIGHT"
     misc_feature    6616..6639)
                     /label=NLS
                     /note="color: #ff00ff"
     CDS             6682..6705)
                     /label=Flag tag
                     /note="color: #008000; direction: RIGHT"
     misc_feature    6712..6980)
                     /label=CYC1 terminator
                     /note="note=""direct strand 
                     translation""translation=""SLLESCN*"""
                     /note="color: #0000ff"
     rep_origin      complement(7031..7486))
                     /direction=LEFT
                     /label=M13 origin
                     /note="color: #4682b4"
     rep_origin      7041..7481)
                     /direction=RIGHT
                     /label=F1 ori
                     /note="color: #4682b4"
     CDS             complement(7491..7559))
                     /label=LacZ alpha
                     /note="color: #6495ed; direction: LEFT"
     primer_bind     7630..7647)
                     /label=M13-fwd
                     /note="color: #008b8b; direction: RIGHT"
     primer_bind     7654..7673)
                     /label=T7
                     /note="color: #008b8b; direction: RIGHT"
     misc_feature    7753..8021)
                     /label=SNR52p
                     /note="color: #ffff00"
     misc_feature    8022..8048)
                     /label=BplI
                     /note="color: #ff0000"
     misc_feature    8049..8147)
                     /label=sup4 term + tracer
                     /note="color: #00ff00"
     primer_bind     complement(8188..8207))
                     /label=T3
                     /note="color: #008b8b; direction: LEFT"
     primer_bind     complement(8225..8245))
                     /label=M13-rev
                     /note="color: #008b8b; direction: LEFT"
     rep_origin      complement(8613..9241))
                     /direction=LEFT
                     /label=ColE1 origin
                     /note="color: #4682b4"
     CDS             complement(9393..10052))
                     /label=AmpR
                     /note="color: #8fbc8f; direction: LEFT"
     promoter        complement(10292..10320))
                     /label=Amp prom
                     /note="color: #4682b4; direction: LEFT"
     rep_origin      10382..10892)
                     /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 gggacgaggc aagctaaaca gatctgtaaa gagccccatt
     1921 atcttagcct aaaaaaacct tctctttgga actttcagta atacgcttaa ctgctcattg
     1981 ctatattgaa gtacggatta gaagccgccg agcgggtgac agccctccga aggaagactc
     2041 tcctccgtgc gtcctcgtct tcaccggtcg cgttcctgaa acgcagatgt gcctcgcgcc
     2101 gcactgctcc gaacaataaa gattctacaa tactagcttt tatggttatg aagaggaaaa
     2161 attggcagta acctggcccc acaaaccttc aaatgaacga atcaaattaa caaccatagg
     2221 atgataatgc gattagtttt ttagccttat ttctggggta attaatcagc gaagcgatga
     2281 tttttgatct attaacagat atataaatgc aaaaactgca taaccacttt aactaatact
     2341 ttcaacattt tcggtttgta ttacttctta ttcaaatgta ataaaagtat caacaaaaaa
     2401 ttgttaatat acctctatac tttaacgtca aggagaaaaa acccggatct caaaatggat
     2461 aaaaagtata gtattggttt agatattggt actaactctg tgggttgggc agttatcact
     2521 gacgaatata aagttccatc aaagaaattt aaggtgttag gtaacactga cagacactca
     2581 ataaaaaaga atcttatcgg tgctcttttg ttcgactccg gtgaaactgc cgaggctaca
     2641 cgtttaaaaa gaacagcaag aagaagatat acccgtagaa aaaatagaat atgttattta
     2701 caagaaatct tttctaatga aatggctaaa gttgatgatt cctttttcca tagattggaa
     2761 gagtcatttt tggttgaaga agacaaaaag catgagagac atccaatctt tgggaatata
     2821 gttgatgaag tggcttacca tgaaaaatat cctaccattt atcatttaag aaagaaattg
     2881 gtagattcaa ctgataaagc tgaccttaga ttaatctatt tagcacttgc ccatatgatt
     2941 aaatttagag gtcatttttt gattgaaggt gatttgaacc cagataattc tgacgtggat
     3001 aaattattta ttcaattagt ccaaacctac aaccaattat ttgaggaaaa tccaattaat
     3061 gctagtggtg tcgatgccaa agctatatta tcagccagat tatcaaaatc tagacgtttg
     3121 gaaaatttga ttgcccaatt gccaggagaa aaaaagaatg gattatttgg aaacttgatc
     3181 gcattatcat tgggtttgac accaaatttt aaatctaatt ttgatttagc tgaagatgct
     3241 aaattacaat tatcaaaaga cacctatgac gacgatttgg acaatttact tgctcaaatt
     3301 ggtgatcaat atgcagattt gttcttagct gctaaaaact tatctgatgc tattttgttg
     3361 tctgatattt tgagagtgaa cacagaaata accaaagctc cattatcagc atctatgatc
     3421 aaacgttatg atgaacacca tcaggatttg actttattga aagctttggt gagacaacaa
     3481 ttgccagaga agtataaaga aatctttttc gatcaatcta aaaacgggta tgcaggttat
     3541 attgatgggg gtgcctccca agaggaattt tacaaattta taaaacctat tttagaaaag
     3601 atggatggga ctgaggaact tttggtcaaa ttgaacagag aagatttgtt acgtaaacag
     3661 agaacttttg ataatggtag tatacctcac caaattcatt tgggtgagtt gcatgcaatt
     3721 ttaagaagac aagaagattt ttatccattt ttaaaagata atagagaaaa aatcgagaaa
     3781 attttaacct ttagaattcc atactatgtt gggcctttgg ctagaggtaa ttcaagattt
     3841 gcctggatga cacgtaaatc agaagaaact attacccctt ggaattttga agaggttgtt
     3901 gataaaggag catcagcaca gagttttatt gaaagaatga ccaatttcga taaaaactta
     3961 ccaaatgaaa aagttttacc aaaacattcc ttgttatacg aatattttac tgtttacaat
     4021 gaacttacaa aggttaaata tgttactgaa ggtatgcgta agccagcctt tttatctgga
     4081 gaacagaaaa aggcaatagt tgatttattg tttaaaacaa atagaaaagt tactgttaaa
     4141 caattaaaag aagattactt taagaaaatt gaatgttttg attcagttga aatcagtggt
     4201 gttgaagaca gatttaatgc tagtttagga acttaccatg atttacttaa aattatcaaa
     4261 gataaagatt tcttggataa cgaagaaaat gaagacattt tagaagacat tgttttaacc
     4321 ttaactttat tcgaagatag agagatgatt gaagaacgtt tgaagactta tgcacatttg
     4381 tttgacgata aagtgatgaa acagttgaaa agaagacgtt atactggatg gggtagattg
     4441 tctcgtaaat tgatcaatgg aattagagat aaacaaagtg gtaaaactat cttggacttt
     4501 ttgaaatctg acggatttgc taatagaaat ttcatgcaat tgatccacga cgatagtttg
     4561 acatttaaag aagacatcca aaaggcccaa gtgagtgggc aaggtgattc attacatgaa
     4621 catattgcaa atttagccgg atctcctgct attaagaaag ggatattaca aactgttaaa
     4681 gttgtggatg aattagtgaa agtaatggga agacataaac ctgaaaacat tgtcattgag
     4741 atggcaagag aaaatcaaac tacacaaaaa ggacagaaaa atagtagaga acgtatgaaa
     4801 agaatagaag agggtattaa agaattgggt agtcaaatat tgaaagaaca cccagtggaa
     4861 aatacccagt tgcaaaatga aaaattatat ctttactacc ttcaaaatgg acgtgatatg
     4921 tatgttgatc aggaattaga tataaataga ctttcagatt atgatgtaga tcatatagtt
     4981 ccacaatctt tcttgaaaga tgattccata gacaataaag tattaactag aagtgataaa
     5041 aatagaggta aaagtgataa tgtcccaagt gaggaagtcg tcaaaaagat gaaaaattac
     5101 tggcgtcaac ttttgaatgc taaattaatt actcaaagaa aatttgataa tttgactaaa
     5161 gcagaaagag gtgggctttc tgaattagat aaagccgggt tcattaaaag acaattggtc
     5221 gaaactagac aaattactaa acatgttgcc caaattttag attcccgtat gaacactaag
     5281 tatgacgaaa atgataagtt aatacgtgag gttaaagtca ttactttaaa atcaaaactt
     5341 gtctctgatt tcagaaagga tttccaattc tataaagtta gagaaattaa taattatcat
     5401 catgctcatg atgcatattt gaatgctgta gttggaactg ctttaatcaa gaaataccct
     5461 aaattagaat ctgaatttgt atatggtgat tacaaagtct atgatgttag aaagatgatt
     5521 gctaaatcag aacaagaaat tggtaaagct acagctaaat acttctttta ctctaacatt
     5581 atgaatttct ttaaaacaga aattactttg gcaaacggtg aaattagaaa aagacctctt
     5641 attgaaacaa atggtgagac tggagagata gtttgggaca aagggcgtga tttcgctact
     5701 gttagaaaag ttttatcaat gccacaagtt aacattgtaa agaaaacaga ggttcaaact
     5761 ggtggtttct caaaagaaag tattttgcct aaaagaaata gtgataaatt gattgccaga
     5821 aaaaaggatt gggatccaaa gaaatatggt ggtttcgact caccaaccgt agcctattct
     5881 gttttggttg tggcaaaggt tgaaaagggt aaaagtaaaa agcttaaatc agtaaaagaa
     5941 cttttgggta ttacaataat ggaaagaagt tcctttgaaa agaaccctat tgattttttg
     6001 gaagctaaag gttataagga agtaaagaag gacttaataa tcaaattgcc taaatattct
     6061 ttatttgaat tagaaaatgg gagaaaaaga atgttggctt ctgctggaga attgcaaaag
     6121 ggtaatgaat tagcattgcc ttccaaatat gttaacttct tgtatttagc ttcacactat
     6181 gaaaagttga aagggtcacc agaagataac gagcaaaaac aattatttgt tgaacaacac
     6241 aaacactact tagatgagat tatagaacaa attagtgaat tcagtaaaag agtgatatta
     6301 gctgatgcaa atttagataa agttttgtca gcctataaca aacatagaga taagccaatt
     6361 agagaacaag cagaaaacat tattcactta tttaccctta ccaatttagg agcacctgct
     6421 gctttcaagt attttgatac aacaattgat cgtaaaagat atacctcaac aaaagaagtc
     6481 ttagacgcca ccttaattca tcaatcaatc actggattgt atgagacaag aattgatttg
     6541 tctcaattgg gtggtgatga aggggctgat cctaagaaga aaagaaaagt tgatccaaag
     6601 aaaaagcgta aggtggatcc taagaaaaag agaaaggttg actacaaaga ccatgacggt
     6661 gattataaag atcatgacat cgactacaag gatgacgatg acaagtgata atctcttctc
     6721 gagtcatgta attagttatg tcacgcttac attcacgccc tccccccaca tccgctctaa
     6781 ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta
     6841 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg
     6901 tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg
     6961 ctttaatttg cggccggtac tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     7021 ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     7081 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     7141 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     7201 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     7261 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     7321 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     7381 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     7441 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcgcg ccattcgcca
     7501 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     7561 ctggcgaagg ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     7621 tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg
     7681 gagctccacc gcggtggcgg ccgctctaga actagtggat cccccgggct gcaggaattc
     7741 gatatcaagc tttctttgaa aagataatgt atgattatgc tttcactcat atttatacag
     7801 aaacttgatg ttttctttcg agtatataca aggtgattac atgtacgttt gaagtacaac
     7861 tctagatttt gtagtgccct cttgggctag cggtaaaggt gcgcattttt tcacacccta
     7921 caatgttctg ttcaaaagat tttggtcaaa cgctgtagaa gtgaaagttg gtgcgcatgt
     7981 ttcggcgttc gaaacttctc cgcagtgaaa gataaatgat cctagatgcg agatatcctc
     8041 atgcctaggt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
     8101 gaaaaagtgg caccgagtcg gtggtgcttt ttttgttttt tatgtctacc gtcgacctcg
     8161 agggggggcc cggtacccag cttttgttcc ctttagtgag ggttaattcc gagcttggcg
     8221 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     8281 ataggagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag gtaactcaca
     8341 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     8401 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     8461 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     8521 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     8581 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     8641 ctcggccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     8701 acaggactat aaagatacca ggcgttcccc cctggaagct ccctcgtgcg ctctcctgtt
     8761 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     8821 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     8881 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     8941 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     9001 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     9061 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     9121 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     9181 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     9241 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     9301 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     9361 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     9421 tcagcgatct gtctatttcg ttcatccata gttgcctgac tgcccgtcgt gtagataact
     9481 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     9541 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     9601 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     9661 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     9721 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     9781 acatgatccc ccatgttgtg aaaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     9841 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     9901 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     9961 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
    10021 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
    10081 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
    10141 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
    10201 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
    10261 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
    10321 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
    10381 gggtcctttt catcacgtgc tataaaaata attataattt aaatttttta atataaatat
    10441 ataaattaaa aatagaaagt aaaaaaagaa attaaagaaa aaatagtttt tgttttccga
    10501 agatgtaaaa gactctaggg ggatcgccaa caaatactac cttttatctt gctcttcctg
    10561 ctctcaggta ttaatgccga attgtttcat cttgtctgtg tagaagacca cacacgaaaa
    10621 tcctgtgatt ttacatttta cttatcgtta atcgaatgta tatctattta atctgctttt
    10681 cttgtctaat aaatatatat gtaaagtacg ctttttgttg aaatttttta aacctttgtt
    10741 tatttttttt tcttcattcc gtaactcttc taccttcttt atttactttc taaaatccaa
    10801 atacaaaaca taaaaataaa taaacacaga gtaaattccc aaattattcc atcattaaaa
    10861 gatacgaggc gcgtgtaagt tacaggcaag cgatccgtcc taagaaacca ttattatcat
    10921 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtc
//
