LOCUS       pTO135_C._auris_pENO1-C 8226 bp ds-DNA     circular     15-JUL-2020
DEFINITION  .
FEATURES             Location/Qualifiers
     CDS             complement(239..307)
                     /label="LacZ alpha"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer_bind     378..395
                     /label="M13-fwd"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer          379..394
                     /label="oTO19"
                     /note="sequence: GTAAAACGACGGCCAG"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    complement(396..413)
                     /label="rev"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     primer          complement(396..413)
                     /label="oTO100 CauElm1KO-Gibs-pUC19-R"
                     /note="sequence: tttcgccaaacttgcGGTACCGAGCTCGAATTC"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     primer          complement(396..428)
                     /label="oTO117 pUC19 Backbone_rev_pTO135"
                     /note="sequence: tgaactacaaagaggGGTACCGAGCTCGAATTC"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     primer          399..435
                     /label="oTO112 pENO1_fwd_pTO135"
                     /note="sequence: ttcgagctcggtaccCCTCTTTGTAGTTCAACTTATG"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    414..436
                     /label="oTO143"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     primer          414..436
                     /label="oTO143 Cau_Cas9_fwd"
                     /note="sequence: CCTCTTTGTAGTTCAACTTATGC"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    414..1413
                     /label="pENO1"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    414..5956
                     /label="Amplified Cas9 for Transient CRISPR"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     primer          complement(1391..1428)
                     /label="oTO113 pENO1_rev_pTO135"
                     /note="sequence: atactttttatccatGATGAAAATTAAGTTTGGATAGG"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer          1399..1441
                     /label="oTO114 Cas9-CYC1term_fwd_pTO135"
                     /note="sequence: aacttaattttcatcATGGATAAAAAGTATAGTATTGGTTTAG"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    1414..5670
                     /label="Ca-Recoded-Cas9-3xNLS"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    5527..5598
                     /label="3x SV40 NLS"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    5729..5956
                     /label="Tcyc1"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     primer          complement(5937..5956)
                     /label="oTO41 CaCAS9-R"
                     /note="sequence: gtcccaaaaccttctcaagc"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     primer          complement(5938..5971)
                     /label="oTO115 Cas9-CYC1term_rev_pTO135"
                     /note="sequence: gactctagaggatccGTCCCAAAACCTTCTCAAG"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     primer          5942..5974
                     /label="oTO116 pUC19 Backbone_fwd_pTO135"
                     /note="sequence: agaaggttttgggacGGATCCTCTAGAGTCGAC"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer          5956..5974
                     /label="oTO99 CauElm1KO-Gibs-pUC19-F"
                     /note="sequence: ttaataacgcccacgGGATCCTCTAGAGTCGAC"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    5957..5974
                     /label="fwd"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer_bind     complement(6001..6021)
                     /label="M13-rev"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer          complement(6005..6021)
                     /label="oTO18"
                     /note="sequence: CAGGAAACAGCTATGAC"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_binding    complement(6027..6049)
                     /label="LacO"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     promoter        complement(6053..6083)
                     /label="lac promoter"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     rep_origin      complement(6389..7071)
                     /label="ColE1 origin"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     CDS             complement(7169..7828)
                     /label="AmpR"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
ORIGIN
        1 TCGCGCGTTT CGGTGATGAC GGTGAAAACC TCTGACACAT GCAGCTCCCG GAGACGGTCA
       61 CAGCTTGTCT GTAAGCGGAT GCCGGGAGCA GACAAGCCCG TCAGGGCGCG TCAGCGGGTG
      121 TTGGCGGGTG TCGGGGCTGG CTTAACTATG CGGCATCAGA GCAGATTGTA CTGAGAGTGC
      181 ACCATATGCG GTGTGAAATA CCGCACAGAT GCGTAAGGAG AAAATACCGC ATCAGGCGCC
      241 ATTCGCCATT CAGGCTGCGC AACTGTTGGG AAGGGCGATC GGTGCGGGCC TCTTCGCTAT
      301 TACGCCAGCT GGCGAAAGGG GGATGTGCTG CAAGGCGATT AAGTTGGGTA ACGCCAGGGT
      361 TTTCCCAGTC ACGACGTTGT AAAACGACGG CCAGTGAATT CGAGCTCGGT ACCCCTCTTT
      421 GTAGTTCAAC TTATGCTCAC TGCTCTTCCC TCCAATACCA TCTGTTCTAC CATCTGCTCT
      481 GCCCTAATTA AACTCACTCA CCTGTCGAAC AGCACCTGTC AAAACAGGTG ACTCCACTAA
      541 TAATACAAAT CCACGTCAAA TGACGTGCAC GAGGGAGTAT GTCATATTCT ATGGCCGTTA
      601 GTCTCCCGGT CAAATTGACC AATCACAAGA CTTACCTGAA GACCTGACTC AACGCAATCG
      661 CCAGTCCCGG GTTTCCTGGC TCACGGGCCC CTCCACTACG CTGATATTTT GTCAATTAAC
      721 CAGGAGTCTA CCCTAGTGCG CATTGTCGAA GGGAGTGGAA AGCCAGGGAA AAAAAACGTA
      781 TAGTGCGGTC CCCGTCGACC CGAGACGACA AGGCACCCAG GTGAGTCAGG CTCAAATGCA
      841 TGAAAAATTA TGAAAAGTGG AAAATCGATG GCCCTCAGGT GAAGGTTATC AAAACCTCAT
      901 CAGCTCGTCG CTCAATTGCT AAAAATCAAC TGAAACAGCG TCCCAGTGGT GTCACCCCAC
      961 GGTTGTTTCA GGGCTGCGGT GGATCACATG GCGACTCCCT TTGGCCCGTA GGGTGCGATC
     1021 CGTCAGTGCC CGGGGCCCTC TGCCAGGATT CTACGCGCAT TGCAACTCCG AGGCCGATAC
     1081 CCTTTGACCA GTGGTCAATT CGTCACAATT GGCAACATTT TCATGATTTG GCAGCTTCAC
     1141 GTTGAACAAT TGAACCAGCA GAATCCAGGA AACGAGTTTC CGTTGTGAAT GACCCTTTGT
     1201 GTGTGCAAAT AATCATGGTG GCTCCCTAAT GGCATACCCT CAGTCACCTG GAGGAATTGC
     1261 TCGCTCGAGT CTCCTGAGGT GTCCTCCTGC CTGTGGAAAA AGCGGCCAGG TGTGGTCCAC
     1321 CGTTGGAACA GCTTGCTGTA TAAATACCAC AGGTGCGCAC GCCCCAGGGG CCTTTTTTTT
     1381 TTTTCTCCTC CCTATCCAAA CTTAATTTTC ATCatggata aaaagtatag tattggttta
     1441 gatattggta ctaactctgt gggttgggca gttatcaccg acgaatataa agttccatca
     1501 aagaaattta aggtgttagg taacactgac agacactcaa taaaaaagaa tcttatcggt
     1561 gctcttttgt tcgactccgg tgaaactgcc gaggctacac gtttaaaaag aacagcaaga
     1621 agaagatata cccgtagaaa aaatagaata tgttatttac aagaaatctt ttctaatgaa
     1681 atggctaaag ttgatgattc ctttttccat agattggaag agtcattttt ggttgaagaa
     1741 gacaaaaagc atgagagaca tccaatcttt gggaatatag ttgatgaagt ggcttaccat
     1801 gaaaaatatc ctaccattta tcatttaaga aagaaattgg tagattcaac tgataaagct
     1861 gaccttagat taatctattt agcacttgcc catatgatta aatttagagg tcattttttg
     1921 attgaaggtg atttgaaccc agataattct gacgtggata aattatttat tcaattagtc
     1981 caaacctaca accaattatt tgaggaaaat ccaattaatg ctagtggtgt cgatgccaaa
     2041 gctatattat cagccagatt atcaaaatct agacgtttgg aaaatttgat tgcccaattg
     2101 ccaggagaaa aaaagaatgg attatttgga aacttgatcg cattatcatt gggtttgaca
     2161 ccaaatttta aatctaattt tgatttagct gaagatgcta aattacaatt atcaaaagac
     2221 acctatgacg acgatttgga caatttactt gctcaaattg gtgatcaata tgcagatttg
     2281 ttcttagctg ctaaaaactt atctgatgct attttgttgt ctgatatttt gagagtgaac
     2341 acagaaataa ccaaagctcc attatcagca tctatgatca aacgttatga tgaacaccat
     2401 caggatttga ctttattgaa agctttggtg agacaacaat tgccagagaa gtataaagaa
     2461 atctttttcg atcaatctaa aaacgggtat gcaggttata ttgatggggg tgcctcccaa
     2521 gaggaatttt acaaatttat aaaacctatt ttagaaaaga tggatgggac tgaggaactt
     2581 ttggtcaaat tgaacagaga agatttgtta cgtaaacaga gaacttttga taatggtagt
     2641 atacctcacc aaattcattt gggtgagttg catgcaattt taagaagaca agaagatttt
     2701 tatccatttt taaaagataa tagagaaaaa atcgagaaaa ttttaacctt tagaattcca
     2761 tactatgttg ggcctttggc tagaggtaat tcaagatttg cctggatgac acgtaaatca
     2821 gaagaaacta ttaccccttg gaattttgaa gaggttgttg ataaaggagc atcagcacag
     2881 agttttattg aaagaatgac caatttcgat aaaaacttac caaatgaaaa agttttacca
     2941 aaacattcct tgttatacga atattttact gtttacaatg aacttacaaa ggttaaatat
     3001 gttactgaag gtatgcgtaa gccagccttt ttatctggag aacagaaaaa ggcaatagtt
     3061 gatttattgt ttaaaacaaa tagaaaagtt actgttaaac aattaaaaga agattacttt
     3121 aagaaaattg aatgttttga ttcagttgaa atcagtggtg ttgaagacag atttaatgct
     3181 agtttaggaa cttaccatga tttacttaaa attatcaaag ataaagattt cttggataac
     3241 gaagaaaatg aagacatttt agaagacatt gttttaacct taactttatt cgaagataga
     3301 gagatgattg aagaacgttt gaagacttat gcacatttgt ttgacgataa agtgatgaaa
     3361 cagttgaaaa gaagacgtta tactggatgg ggtagattgt ctcgtaaatt gatcaatgga
     3421 attagagata aacaaagtgg taaaactatc ttggactttt tgaaatctga cggatttgct
     3481 aatagaaatt tcatgcaatt gatccacgac gatagtttga catttaaaga agacatccaa
     3541 aaggcccaag tgagtgggca aggtgattca ttacatgaac atattgcaaa tttagccgga
     3601 tctcctgcta ttaagaaagg gatattacaa actgttaaag ttgtggatga attagtgaaa
     3661 gtaatgggaa gacataaacc tgaaaacatt gtcattgaga tggcaagaga aaatcaaact
     3721 acacaaaaag gacagaaaaa tagtagagaa cgtatgaaaa gaatagaaga gggtattaaa
     3781 gaattgggta gtcaaatatt gaaagaacac ccagtggaaa atacccagtt gcaaaatgaa
     3841 aaattatatc tttactacct tcaaaatgga cgtgatatgt atgttgatca ggaattagat
     3901 ataaatagac tttcagatta tgatgtagat catatagttc cacaatcttt cttgaaagat
     3961 gattccatag acaataaagt attaactaga agtgataaaa atagaggtaa aagtgataat
     4021 gtcccaagtg aggaagtcgt caaaaagatg aaaaattact ggcgtcaact tttgaatgct
     4081 aaattaatta ctcaaagaaa atttgataat ttgactaaag cagaaagagg tgggctttct
     4141 gaattagata aagccgggtt cattaaaaga caattggtcg aaactagaca aattactaaa
     4201 catgttgccc aaattttaga ttcccgtatg aacactaagt atgacgaaaa tgataagtta
     4261 atacgtgagg ttaaagtcat tactttaaaa tcaaaacttg tctctgattt cagaaaggat
     4321 ttccaattct ataaagttag agaaattaat aattatcatc atgctcatga tgcatatttg
     4381 aatgctgtag ttggaactgc tttaatcaag aaatacccta aattagaatc tgaatttgta
     4441 tatggtgatt acaaagtcta tgatgttaga aagatgattg ctaaatcaga acaagaaatt
     4501 ggtaaagcta cagctaaata cttcttttac tctaacatta tgaatttctt taaaacagaa
     4561 attactttgg caaacggtga aattagaaaa agacctctta ttgaaacaaa tggtgagact
     4621 ggagagatag tttgggacaa agggcgtgat ttcgctactg ttagaaaagt tttatcaatg
     4681 ccacaagtta acattgtaaa gaaaacagag gttcaaactg gtggtttctc aaaagaaagt
     4741 attttgccta aaagaaatag tgataaattg attgccagaa aaaaggattg ggatccaaag
     4801 aaatatggtg gtttcgactc accaaccgta gcctattctg ttttggttgt ggcaaaggtt
     4861 gaaaagggta aaagtaaaaa gcttaaatca gtaaaagaac ttttgggtat tacaataatg
     4921 gaaagaagtt cctttgaaaa gaaccctatt gattttttgg aagctaaagg ttataaggaa
     4981 gtaaagaagg acttaataat caaattgcct aaatattctt tatttgaatt agaaaatggg
     5041 agaaaaagaa tgttggcttc tgctggagaa ttgcaaaagg gtaatgaatt agcattgcct
     5101 tccaaatatg ttaacttctt gtatttagct tcacactatg aaaagttgaa agggtcacca
     5161 gaagataacg agcaaaaaca attatttgtt gaacaacaca aacactactt agatgagatt
     5221 atagaacaaa ttagtgaatt cagtaaaaga gtgatattag ctgatgcaaa tttagataaa
     5281 gttttgtcag cctataacaa acatagagat aagccaatta gagaacaagc agaaaacatt
     5341 attcacttat ttacccttac caatttagga gcacctgctg ctttcaagta ttttgataca
     5401 acaattgatc gtaaaagata tacctcaaca aaagaagtct tagacgccac cttaattcat
     5461 caatcaatca ctggattgta tgagacaaga attgatttgt ctcaattggg tggtgatgaa
     5521 ggggctgatc ctaagaagaa aagaaaagtt gatccaaaga aaaagcgtaa ggtggatcct
     5581 aagaaaaaga gaaaggttga ctacaaagac catgacggtg attataaaga tcatgacatc
     5641 gactacaagg atgacgatga caagtgataa tgactgcaga gatccatcga cctgccgcca
     5701 agctaattcc gggcgaattt ctgtcgagtc atgtaattag ttatgtcacg cttacattca
     5761 cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
     5821 gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt
     5881 tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt
     5941 gagaaggttt tgggacGGAT CCTCTAGAGT CGACCTGCAG GCATGCAAGC TTGGCGTAAT
     6001 CATGGTCATA GCTGTTTCCT GTGTGAAATT GTTATCCGCT CACAATTCCA CACAACATAC
     6061 GAGCCGGAAG CATAAAGTGT AAAGCCTGGG GTGCCTAATG AGTGAGCTAA CTCACATTAA
     6121 TTGCGTTGCG CTCACTGCCC GCTTTCCAGT CGGGAAACCT GTCGTGCCAG CTGCATTAAT
     6181 GAATCGGCCA ACGCGCGGGG AGAGGCGGTT TGCGTATTGG GCGCTCTTCC GCTTCCTCGC
     6241 TCACTGACTC GCTGCGCTCG GTCGTTCGGC TGCGGCGAGC GGTATCAGCT CACTCAAAGG
     6301 CGGTAATACG GTTATCCACA GAATCAGGGG ATAACGCAGG AAAGAACATG TGAGCAAAAG
     6361 GCCAGCAAAA GGCCAGGAAC CGTAAAAAGG CCGCGTTGCT GGCGTTTTTC CATAGGCTCC
     6421 GCCCCCCTGA CGAGCATCAC AAAAATCGAC GCTCAAGTCA GAGGTGGCGA AACCCGACAG
     6481 GACTATAAAG ATACCAGGCG TTTCCCCCTG GAAGCTCCCT CGTGCGCTCT CCTGTTCCGA
     6541 CCCTGCCGCT TACCGGATAC CTGTCCGCCT TTCTCCCTTC GGGAAGCGTG GCGCTTTCTC
     6601 ATAGCTCACG CTGTAGGTAT CTCAGTTCGG TGTAGGTCGT TCGCTCCAAG CTGGGCTGTG
     6661 TGCACGAACC CCCCGTTCAG CCCGACCGCT GCGCCTTATC CGGTAACTAT CGTCTTGAGT
     6721 CCAACCCGGT AAGACACGAC TTATCGCCAC TGGCAGCAGC CACTGGTAAC AGGATTAGCA
     6781 GAGCGAGGTA TGTAGGCGGT GCTACAGAGT TCTTGAAGTG GTGGCCTAAC TACGGCTACA
     6841 CTAGAAGAAC AGTATTTGGT ATCTGCGCTC TGCTGAAGCC AGTTACCTTC GGAAAAAGAG
     6901 TTGGTAGCTC TTGATCCGGC AAACAAACCA CCGCTGGTAG CGGTGGTTTT TTTGTTTGCA
     6961 AGCAGCAGAT TACGCGCAGA AAAAAAGGAT CTCAAGAAGA TCCTTTGATC TTTTCTACGG
     7021 GGTCTGACGC TCAGTGGAAC GAAAACTCAC GTTAAGGGAT TTTGGTCATG AGATTATCAA
     7081 AAAGGATCTT CACCTAGATC CTTTTAAATT AAAAATGAAG TTTTAAATCA ATCTAAAGTA
     7141 TATATGAGTA AACTTGGTCT GACAGTTACC AATGCTTAAT CAGTGAGGCA CCTATCTCAG
     7201 CGATCTGTCT ATTTCGTTCA TCCATAGTTG CCTGACTCCC CGTCGTGTAG ATAACTACGA
     7261 TACGGGAGGG CTTACCATCT GGCCCCAGTG CTGCAATGAT ACCGCGAGAC CCACGCTCAC
     7321 CGGCTCCAGA TTTATCAGCA ATAAACCAGC CAGCCGGAAG GGCCGAGCGC AGAAGTGGTC
     7381 CTGCAACTTT ATCCGCCTCC ATCCAGTCTA TTAATTGTTG CCGGGAAGCT AGAGTAAGTA
     7441 GTTCGCCAGT TAATAGTTTG CGCAACGTTG TTGCCATTGC TACAGGCATC GTGGTGTCAC
     7501 GCTCGTCGTT TGGTATGGCT TCATTCAGCT CCGGTTCCCA ACGATCAAGG CGAGTTACAT
     7561 GATCCCCCAT GTTGTGCAAA AAAGCGGTTA GCTCCTTCGG TCCTCCGATC GTTGTCAGAA
     7621 GTAAGTTGGC CGCAGTGTTA TCACTCATGG TTATGGCAGC ACTGCATAAT TCTCTTACTG
     7681 TCATGCCATC CGTAAGATGC TTTTCTGTGA CTGGTGAGTA CTCAACCAAG TCATTCTGAG
     7741 AATAGTGTAT GCGGCGACCG AGTTGCTCTT GCCCGGCGTC AATACGGGAT AATACCGCGC
     7801 CACATAGCAG AACTTTAAAA GTGCTCATCA TTGGAAAACG TTCTTCGGGG CGAAAACTCT
     7861 CAAGGATCTT ACCGCTGTTG AGATCCAGTT CGATGTAACC CACTCGTGCA CCCAACTGAT
     7921 CTTCAGCATC TTTTACTTTC ACCAGCGTTT CTGGGTGAGC AAAAACAGGA AGGCAAAATG
     7981 CCGCAAAAAA GGGAATAAGG GCGACACGGA AATGTTGAAT ACTCATACTC TTCCTTTTTC
     8041 AATATTATTG AAGCATTTAT CAGGGTTATT GTCTCATGAG CGGATACATA TTTGAATGTA
     8101 TTTAGAAAAA TAAACAAATA GGGGTTCCGC GCACATTTCC CCGAAAAGTG CCACCTGACG
     8161 TCTAAGAAAC CATTATTATC ATGACATTAA CCTATAAAAA TAGGCGTATC ACGAGGCCCT
     8221 TTCGTC
//