LOCUS pWS158_[o] 11081 bp ds-DNA circular 26-JUN-2016 DEFINITION . KEYWORDS "E.coli Marker:KanR" "Type:o" "Yeast Marker:URA3" "Yeast Vector:2micron" FEATURES Location/Qualifiers terminator 6784..6858 /label="tSNR52" /ApEinfo_revcolor=#d59687 /ApEinfo_fwdcolor=#d59687 rep_origin 8314..9276 /label="2micron" /ApEinfo_revcolor=#d6b295 /ApEinfo_fwdcolor=#d6b295 misc_feature 5178..5416 /label="Junk for Gap Repair" /ApEinfo_revcolor=#ffef86 /ApEinfo_fwdcolor=#ffef86 misc_feature 28..52 /label="ConS scar" /ApEinfo_revcolor=#ffef86 /ApEinfo_fwdcolor=#ffef86 rep_origin 9277..9314 /label="FRT site" /ApEinfo_revcolor=#b4abac /ApEinfo_fwdcolor=#b4abac CDS complement(9354..10169) /label="KanR" /ApEinfo_revcolor=#008000 /ApEinfo_fwdcolor=#008000 promoter 5417..5516 /label="tRNA Phe Promoter" /ApEinfo_revcolor=#faac61 /ApEinfo_fwdcolor=#faac61 rep_origin complement(10318..11081) /label="ColE1" /ApEinfo_revcolor=#7f7f7f /ApEinfo_fwdcolor=#7f7f7f RBS 5815..5850 /label="sfGFP Ribosome Binding Site" /ApEinfo_revcolor=#b1ff67 /ApEinfo_fwdcolor=#b1ff67 terminator complement(9327..9353) /label="KanR Terminator" /ApEinfo_revcolor=#b1ff67 /ApEinfo_fwdcolor=#b1ff67 promoter complement(10170..10317) /label="KanR Promoter" /ApEinfo_revcolor=#b1ff67 /ApEinfo_fwdcolor=#b1ff67 CDS 766..4866 /label="Cas9" /ApEinfo_revcolor=#85dae9 /ApEinfo_fwdcolor=#85dae9 terminator 4942..5166 /label="ScPGK1 Terminator" /ApEinfo_revcolor=#ff9ccd /ApEinfo_fwdcolor=#ff9ccd CDS 5851..6567 /label="sfGFP" /ApEinfo_revcolor=#b1ff67 /ApEinfo_fwdcolor=#b1ff67 misc_feature 5608..5677 /label="HDV Ribozyme" /ApEinfo_revcolor=#84b0dc /ApEinfo_fwdcolor=#84b0dc sig_peptide 4876..4926 /label="2x NLS Tag" /ApEinfo_revcolor=#85dae9 /ApEinfo_fwdcolor=#85dae9 misc_feature 6859..7203 /label="Junk for Gap Repair" /ApEinfo_revcolor=#ffef86 /ApEinfo_fwdcolor=#ffef86 CDS 7428..8231 /label="URA3" /ApEinfo_revcolor=#faac61 /ApEinfo_fwdcolor=#faac61 terminator 8232..8309 /label="ScURA3 Terminator" /ApEinfo_revcolor=#f8d3a9 /ApEinfo_fwdcolor=#f8d3a9 misc_RNA 6704..6783 /label="sgRNA" /ApEinfo_revcolor=#85dae9 /ApEinfo_fwdcolor=#85dae9 misc_feature 7207..7231 /label="ConE scar" /ApEinfo_revcolor=#ffef86 /ApEinfo_fwdcolor=#ffef86 promoter 5685..5814 /label="BBa_J72163 GlpT Promoter" /ApEinfo_revcolor=#b1ff67 /ApEinfo_fwdcolor=#b1ff67 promoter 57..756 /label="ScPGK1 Promoter" /ApEinfo_revcolor=#84b0dc /ApEinfo_fwdcolor=#84b0dc tRNA 5517..5607 /label="tRNA Phe" /ApEinfo_revcolor=#ff9ccd /ApEinfo_fwdcolor=#ff9ccd promoter 7251..7427 /label="ScURA3 Promoter" /ApEinfo_revcolor=#faac61 /ApEinfo_fwdcolor=#faac61 terminator 6568..6696 /label="BBa_B0015 Terminator" /ApEinfo_revcolor=#b1ff67 /ApEinfo_fwdcolor=#b1ff67 ORIGIN 1 gcggccgccc ctgaattcgc atctagactg aactggccga taattgcaga cgaacggtga 61 gtaaggaaag agtgaggaac tatcgcatac ctgcatttaa agatgccgat ttgggcgcga 121 atcctttatt ttggcttcac cctcatacta ttatcagggc cagaaaaagg aagtgtttcc 181 ctccttcttg aattgatgtt accctcataa agcacgtggc ctcttatcga gaaagaaatt 241 accgtcgctc gtgatttgtt tgcaaaaaga acaaaactga aaaaacccag acacgctcga 301 cttcctgtca tcctattgat tgcagcttcc aatttcgtca cacaacaagg tcctagcgac 361 ggctcacagg ttttgtaaca agcaatcgaa ggttctggaa tggcgggaaa gggtttagta 421 ccacatgcta tgatgcccac tgtgatctcc agagcaaagt tcgttcgatc gtactgttac 481 tctctctctt tcaaacagaa ttgtccgaat cgtgtgacaa caacagcctg ttctcacaca 541 ctcttttctt ctaaccaagg gggtggttta gtttagtaga acctcgtgaa acttacattt 601 acatatatat aaacttgcat aaattggtca atgcaagaaa tacatatttg gtcttttcta 661 attcgtagtt tttcaagttc ttagatgctt tctttttctc ttttttacag atcatcaagg 721 aagtaattat ctacttttta caacaaatat aaaacaagat ctatggacaa gaagtattct 781 atcggactgg atatcgggac taatagcgtc gggtgggccg tgatcactga cgagtacaag 841 gtgccctcta agaagttcaa ggtgctcggg aacaccgacc ggcattccat caagaaaaat 901 ctgatcggag ctctcctctt tgattcaggg gaaaccgctg aagcaacccg cctcaagcgg 961 actgctagac ggcggtacac caggaggaag aaccggattt gttaccttca agagatattc 1021 tccaacgaaa tggcaaaggt cgacgacagc ttcttccata ggctggaaga atcattcctc 1081 gtggaagagg ataagaagca tgaacggcat cccatcttcg gtaatatcgt cgacgaggtg 1141 gcctatcacg agaaataccc aaccatctac catcttcgca aaaagctggt ggactcaacc 1201 gacaaggcag acctccggct tatctacctg gccctggccc acatgatcaa gttcagaggc 1261 cacttcctga tcgagggcga cctcaatcct gacaatagcg atgtggataa actgttcatc 1321 cagctggtgc agacttacaa ccagctcttt gaagagaacc ccatcaatgc aagcggagtc 1381 gatgccaagg ccattctgtc agcccggctg tcaaagagcc gcagacttga gaatcttatc 1441 gctcagctgc cgggtgaaaa gaaaaatgga ctgttcggga acctgattgc tctttcactt 1501 gggctgactc ccaatttcaa gtctaatttc gacctggcag aggatgccaa gctgcaactg 1561 tccaaggaca cctatgatga cgatctcgac aacctcctgg cccagatcgg tgaccaatac 1621 gccgaccttt tccttgctgc taagaatctt tctgacgcca tcctgctgtc tgacattctc 1681 cgcgtgaaca ctgaaatcac caaggcccct ctttcagctt caatgattaa gcggtatgat 1741 gagcaccacc aggacctgac cctgcttaag gcactcgtcc ggcagcagct tccggagaag 1801 tacaaggaaa tcttctttga ccagtcaaag aatggatacg ccggctacat cgacggaggt 1861 gcctcccaag aggaatttta taagtttatc aaacctatcc ttgagaagat ggacggcacc 1921 gaagagctcc tcgtgaaact gaatcgggag gatctgctgc ggaagcagcg cactttcgac 1981 aatgggagca ttccccacca gatccatctt ggggagcttc acgccatcct tcggcgccaa 2041 gaggacttct acccctttct taaggacaac agggagaaga ttgagaaaat tctcactttc 2101 cgcatcccct actacgtggg acccctcgcc agaggaaata gccggtttgc ttggatgacc 2161 agaaagtcag aagaaactat cactccctgg aacttcgaag aggtggtgga caagggagcc 2221 agcgctcagt cattcatcga acggatgact aacttcgata agaacctccc caatgagaag 2281 gtcctgccga aacattccct gctctacgag tactttaccg tgtacaacga gctgaccaag 2341 gtgaaatatg tcaccgaagg gatgaggaag cccgcattcc tgtcaggcga acaaaagaag 2401 gcaattgtgg accttctgtt caagaccaat agaaaggtga ccgtgaagca gctgaaggag 2461 gactatttca agaaaattga atgcttcgac tctgtggaga ttagcggggt cgaagatcgg 2521 ttcaacgcaa gcctgggtac ttaccatgat ctgcttaaga tcatcaagga caaggatttt 2581 ctggacaatg aggagaacga ggacatcctt gaggacattg tcctgactct cactctgttc 2641 gaggaccggg aaatgatcga ggagaggctt aagacctacg cccatctgtt cgacgataaa 2701 gtgatgaagc aacttaaacg gagaagatat accggatggg gacgccttag ccgcaaactc 2761 atcaacggaa tccgggacaa acagagcgga aagaccattc ttgatttcct taagagcgac 2821 ggattcgcta atcgcaactt catgcaactt atccatgatg attccctgac ctttaaggag 2881 gacatccaga aggcccaagt gtctggacaa ggtgactcac tgcacgagca tatcgcaaat 2941 ctggctggtt cacccgctat taagaagggt attctccaga ccgtgaaagt cgtggacgag 3001 ctggtcaagg tgatgggtcg ccataaacca gagaacattg tcatcgagat ggccagggaa 3061 aaccagacta cccagaaggg acagaagaac agcagggagc ggatgaaaag aattgaggaa 3121 gggattaagg agctcgggtc acagatcctt aaagagcacc cggtggaaaa cacccagctt 3181 cagaatgaga agctctatct gtactacctt caaaatggac gcgatatgta tgtggaccaa 3241 gagcttgata tcaacaggct ctcagactac gacgtggacc atatcgtccc tcagagcttc 3301 ctcaaagacg actcaattga caataaggtg ctgactcgct cagacaagaa ccggggaaag 3361 tcagataacg tgccctcaga ggaagtcgtg aaaaagatga agaactattg gcgccagctt 3421 ctgaacgcaa agctgatcac tcagcggaag ttcgacaatc tcactaaggc tgagaggggc 3481 ggactgagcg aactggacaa agcaggattc attaaacggc aacttgtgga gactcggcag 3541 attactaaac atgtcgccca aatccttgac tcacgcatga ataccaagta cgacgaaaac 3601 gacaaactta tccgcgaggt gaaggtgatt accctgaagt ccaagctggt cagcgatttc 3661 agaaaggact ttcaattcta caaagtgcgg gagatcaata actatcatca tgctcatgac 3721 gcatatctga atgccgtggt gggaaccgcc ctgatcaaga agtacccaaa gctggaaagc 3781 gagttcgtgt acggagacta caaggtctac gacgtgcgca agatgattgc caaatctgag 3841 caggagatcg gaaaggccac cgcaaagtac ttcttctaca gcaacatcat gaatttcttc 3901 aagaccgaaa tcacccttgc aaacggtgag atccggaaga ggccgctcat cgagactaat 3961 ggggagactg gcgaaatcgt gtgggacaag ggcagagatt tcgctaccgt gcgcaaagtg 4021 ctttctatgc ctcaagtgaa catcgtgaag aaaaccgagg tgcaaaccgg aggcttttct 4081 aaggaatcaa tcctccccaa gcgcaactcc gacaagctca ttgcaaggaa gaaggattgg 4141 gaccctaaga agtacggcgg attcgattca ccaactgtgg cttattctgt cctggtcgtg 4201 gctaaggtgg aaaaaggaaa gtctaagaag ctcaagagcg tgaaggaact gctgggtatc 4261 accattatgg agcgcagctc cttcgagaag aacccaattg actttctcga agccaaaggt 4321 tacaaggaag tcaagaagga ccttatcatc aagctcccaa agtatagcct gttcgaactg 4381 gagaatgggc ggaagcggat gctcgcctcc gctggcgaac ttcagaaggg taatgagctg 4441 gctctcccct ccaagtacgt gaatttcctc taccttgcaa gccattacga gaagctgaag 4501 gggagccccg aggacaacga gcaaaagcaa ctgtttgtgg agcagcataa gcattatctg 4561 gacgagatca ttgagcagat ttccgagttt tctaaacgcg tcattctcgc tgatgccaac 4621 ctcgataaag tccttagcgc atacaataag cacagagaca aaccaattcg ggagcaggct 4681 gagaatatca tccacctgtt caccctcacc aatcttggtg cccctgccgc attcaagtac 4741 ttcgacacca ccatcgaccg gaaacgctat acctccacca aagaagtgct ggacgccacc 4801 ctcatccacc agagcatcac cggactttac gaaactcgga ttgacctctc acagctcgga 4861 ggggatgagg gagctgatcc aaaaaagaag agaaaggtag atccaaaaaa gaagagaaag 4921 gtataaggat cctaactcga gattgaattg aattgaaatc gatagatcaa tttttttctt 4981 ttctctttcc ccatccttta cgctaaaata atagtttatt ttattttttg aatatttttt 5041 atttatatac gtatatatag actattattt atcttttaat gattattaag atttttatta 5101 aaaaaaaatt cgctcctctt ttaatgcctt tatgcagttt ttttttccca ttcgatattt 5161 ctatgtgctg CCAAGATATC AACAACAGAG GACATATGCC CTACCTCCAT GATCTACTGA 5221 CGTCCCTGAG GCTGCAATTC ATGTAATGGG GCAGTATCCG CGGCAAGTCC TAGTGCAATG 5281 GCGGTTTTTT ACCCTCGTTC TGAAGAAGAG GCGACGCGGG TGCGGTCATC ACTAATGTGG 5341 AAATTGGTAA GACTCTCGGG CCTCCGCCTT TAGGCGGTGC TTACTCTTTC ATAAAGGGGC 5401 TGTTAGTTAT GCaacgatgt gcttcagtat tacatttttt gccttcaacg ccttgattgt 5461 tctatttttg ctaataataa atctatttca tcggactaaa agtccattag ttgtaagcgg 5521 atttagctca gttgggagag cgccagactg aagaaaaact tcggtcaagt catctggagg 5581 tcctgtgttc gatccacaga attcgcagat ggccggcatg gtcccagcct cctcgctggc 5641 gccggctggg caacaccttc gggtggcgaa tgggacttga gacggaaagt gaaacgtgat 5701 ttcatgcgtc attttgaaca ttttgtaaat cttatttaat aatgtgtgcg gcaattcaca 5761 tttaatttat gaatgttttc ttaacatcgc ggcaactcaa gaaacggcag gttcggatct 5821 tagctactag agaaagagga gaaatactag atgcgtaaag gcgaagagct gttcactggt 5881 gtcgtcccta ttctggtgga actggatggt gatgtcaacg gtcataagtt ttccgtgcgt 5941 ggcgagggtg aaggtgacgc aactaatggt aaactgacgc tgaagttcat ctgtactact 6001 ggtaaactgc cggttccttg gccgactctg gtaacgacgc tgacttatgg tgttcagtgc 6061 tttgctcgtt atccggacca tatgaagcag catgacttct tcaagtccgc catgccggaa 6121 ggctatgtgc aggaacgcac gatttccttt aaggatgacg gcacgtacaa aacgcgtgcg 6181 gaagtgaaat ttgaaggcga taccctggta aaccgcattg agctgaaagg cattgacttt 6241 aaagaggacg gcaatatcct gggccataag ctggaataca attttaacag ccacaatgtt 6301 tacatcaccg ccgataaaca aaaaaatggc attaaagcga attttaaaat tcgccacaac 6361 gtggaggatg gcagcgtgca gctggctgat cactaccagc aaaacactcc aatcggtgat 6421 ggtcctgttc tgctgccaga caatcactat ctgagcacgc aaagcgttct gtctaaagat 6481 ccgaacgaga aacgcgatca tatggttctg ctggagttcg taaccgcagc gggcatcacg 6541 catggtatgg atgaactgta caaatgacca ggcatcaaat aaaacgaaag gctcagtcga 6601 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca 6661 ctggctcacc ttcgggtggg cctttctgcg tttatacgtc tcagttttag agctagaaat 6721 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct 6781 ttttttattt tttgtcacta ttgttatgta aaatgccacc tctgacagta tggaacgcaa 6841 acttctgtct agtggatagc tgGACATTCC AGGCAGTGCC TCTGCCGCCG GACCCCTCTC 6901 GTGATTGGGT AGCTGGACAT GCCCTTGTAA GATATAACAA GAGCCTGCCT GTCTAATGAT 6961 CTCACGGCGA AAGTCGGGGA GACAGCAGCG GCTGCAGACA TTATACCGCA ACAACACTAA 7021 GGTGAGATAA CTCCGTACAT TGACTACGCG TTCCTCTAGA CCTTACTTGA CCGGATACAG 7081 TGTCTTTGAC ACGTTTATGG GTTACAGCAA TCACATCCAA GACTGGCTAT GCACGAAGCA 7141 ACTCTTGAGT GTTAAAATGT TGACCCCTGT ATTTGGGATG CGGGTAGTAG ATGAGTGCAG 7201 GATATCagca gttacagaga tgttacgaac cactagtgca ctgcagtaca cggtttcctt 7261 gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag gagcacagac 7321 ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc ccagtattct 7381 taacccaact gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta 7441 catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta tttaatatca 7501 tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc aaggaattac 7561 tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat gtggatatct 7621 tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc gccaagtaca 7681 attttttact cttcgaggac agaaaatttg ctgacattgg taatacagtc aaattgcagt 7741 actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca cacggtgtgg 7801 tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca aaggaaccta 7861 gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact ggagaatata 7921 ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc tttattgctc 7981 aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca cccggtgtgg 8041 gtttagatga caagggagat gcattgggtc aacagtatag aaccgtggat gatgtggttt 8101 ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga agggatgcta 8161 aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga agatgcggcc 8221 agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca caaattagag 8281 cttcaattta attatatcag ttattacccg agtaacgaag catctgtgct tcattttgta 8341 gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt 8401 acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg tgcttcattt 8461 ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca 8521 tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga atctatactt 8581 cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa gcatcttaga 8641 ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt tttgcactgt 8701 aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc ataaaaaaag 8761 cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca ttttttcaag 8821 ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt tgtgaacaga 8881 aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt cttctatttt 8941 gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg attcactcta 9001 tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata aacataaaaa 9061 atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg taggttatat 9121 agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt tgtggaagcg 9181 gtattcgcaa tattttagta gctcgttaca gtccggtgcg tttttggttt tttgaaagtg 9241 cgtcatcaga gcgcttttgg ttttcaaaag cgctctgaag ttcctatact ttctagctag 9301 agaataggaa cttcccgagc ggccgcgtgt tacaaccaat taaccaattc tgattagaaa 9361 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat 9421 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg 9481 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat 9541 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc 9601 ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta 9661 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga 9721 gcgaggcgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac 9781 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct 9841 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga 9901 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg 9961 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct 10021 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg 10081 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctggag 10141 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca 10201 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt 10261 tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagtca 10321 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga 10381 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 10441 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 10501 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt 10561 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 10621 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 10681 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 10741 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 10801 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 10861 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 10921 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 10981 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 11041 tgttctttcc tgcgttatcc cctgattctg tggataaccg t //