LOCUS       pTO136_C._auris_pADH1-t 4075 bp ds-DNA     circular     17-NOV-2020
DEFINITION  .
FEATURES             Location/Qualifiers
     CDS             complement(239..307)
                     /label="LacZ alpha"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    296..312
                     /label="oTO224 1st Round PCR"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     primer          296..312
                     /label="oTO224 CausgRNA_f"
                     /note="sequence: GCTATTACGCCAGCTGG"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     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..428)
                     /label="oTO121 pUC19 Backbone_rev_pTO136"
                     /note="sequence: tttcgatctatctcgGGTACCGAGCTCGAATTC"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     primer          399..432
                     /label="oTO118 pADH1_fwd_pTO136"
                     /note="sequence: ttcgagctcggtaccCGAGATAGATCGAAATACG"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    414..433
                     /label="oTO141"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer          414..433
                     /label="oTO141 Cau_sgRNA_fwd"
                     /note="sequence: CGAGATAGATCGAAATACGC"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    414..1314
                     /label="pADH1"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    414..1805
                     /label="C. auris sgRNA Cassette"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          complement(1295..1344)
                     /label="oTO119 pADH1_rev_pTO136"
                     /note="sequence: cgctaccaactacgccacacgcccgagtagGATTTCGTGAAGATTGATTG"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    1315..1393
                     /label="tRNA-Ala"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    complement(1377..1393)
                     /label="upstream sgRNA primer without overlap"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     primer          complement(1377..1393)
                     /label="oTO276 Cau_Iff/Hyr7sg2up"
                     /note="sequence: TTACAAGGGTATTTATCGAATGGACGAGTCCGGATTC"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          complement(1377..1393)
                     /label="oTO274 Cau_Iff/Hyr3sg2up"
                     /note="sequence: AACGTACCAATTCTCTTGTGTGGACGAGTCCGGATTC"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     misc_feature    1393..1393
                     /label="Insert gRNA 5'"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    1394..1413
                     /label="B8441 ENO1 3' gRNA"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     primer          1412..1436
                     /label="oTO275 Cau_Iff/Hyr3sg2down"
                     /note="sequence: CACAAGAGAATTGGTACGTTgttttagagctagaaatagcaag"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    complement(1414..1414)
                     /label="Insert gRNA 3'"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    1414..1436
                     /label="downstream sgRNA primer without overlap"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     primer          1414..1436
                     /label="oTO277 Cau_Iff/Hyr7sg2down"
                     /note="sequence: TTCGATAAATACCCTTGTAAgttttagagctagaaatagcaag"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    1414..1493
                     /label="sgRNA Template"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     misc_feature    1494..1561
                     /label="HDV Ribozyme"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     misc_feature    1562..1805
                     /label="AgTEF2 Terminator"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer          1703..1721
                     /label="oTO92 Cau-Elm1KO-ntermflank-R"
                     /note="sequence: cagatgcgaagttaagtgc"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer          1776..1823
                     /label="oTO120 pUC19 Backbone_fwd_pTO136"
                     /note="sequence: gtcgattcgatactaacgccgccatccagtGGATCCTCTAGAGTCGAC"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     misc_feature    complement(1788..1804)
                     /label="oTO142"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer          complement(1788..1804)
                     /label="oTO142 Cau_sgRNA_rev"
                     /note="sequence: ctggatggcggcgttag"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer          1805..1823
                     /label="oTO99 CauElm1KO-Gibs-pUC19-F"
                     /note="sequence: ttaataacgcccacgGGATCCTCTAGAGTCGAC"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    1806..1823
                     /label="fwd"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     primer_bind     complement(1850..1870)
                     /label="M13-rev"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer          complement(1854..1870)
                     /label="oTO18"
                     /note="sequence: CAGGAAACAGCTATGAC"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_binding    complement(1876..1898)
                     /label="LacO"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     promoter        complement(1902..1932)
                     /label="lac promoter"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    complement(1955..1974)
                     /label="oTO225 1st Round PCR"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     primer          complement(1955..1974)
                     /label="oTO225 CausgRNA_r"
                     /note="sequence: CGCAATTAATGTGAGTTAGC"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     rep_origin      complement(2238..2920)
                     /label="ColE1 origin"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     CDS             complement(3018..3677)
                     /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 ACCCGAGATA
      421 GATCGAAATA CGCTCTAAAG TTTCCAATAC GATTTAAAAC GCATAAGGTG GTCCGGCAGC
      481 AGGCCCTTCA AAGCGGGAAG TATACTTCTA GAAACTATTC ATTGCGCTAG ACGCTCATAT
      541 GCAACGCCAC TCAAATCTGC AAAATTATTC AGATCTATGC TGACATACTC TGACGATTCT
      601 GTGGCGATGC ACTTTTTTTT TTTTTTTTTT TCATTTTTTT TTTTATTTTT TTTGCAACCG
      661 TTCCATTCGA CAGGTTACAT TTCTAGAAAC GTCTGTCCCC AACATGGCAC GGAAAAATTA
      721 GACCAGATTC CCACGTACTG GGCGACTTGG CTGTGCATGA TGCAGTTTCG GGCATCATGA
      781 TCCTTAAGCG CATACATCAG GTGATGCGAC ACCACATCGT GACGAAAGGA AGCTGGAGCC
      841 AGGCCAATTT CACCGCAGCC TTTGACCCTG AGCCGCAAGA CACCACCCCT CCTTGGCGGC
      901 GATGCCCGGT TCTTCCAATT GCGGCTCGTA GGTGCAGTTG AAACGACGAT CTGATGTTCC
      961 ATGAAAGAGC AGATAGAAAA AAGTGAGATT GGCGAAGACA GTAAAATTTC AACACCACAT
     1021 GTCAATCTTA CAAAAACAAT CAATCAGTTC ATTTTTCTTC TCCCCTCAAC TACAATTTTC
     1081 AAAAATGCCA GTGAATGAAA AAAAAGACCC ATTGTCCCAA AACTGGCTCC CTTCTTGAAA
     1141 CCACCTGATT CCTCGAGCGA ATCGGATGGG AAAAAGCCGC ACCGTCACCC AATGCGAACC
     1201 ACCGGCCATG CGTCAGATGG CACCCTTTGG CTGCGAAAAA TATAAAAGCA GAGCAACACC
     1261 GGTGGAATTT CCAAAGTCTT TTCTTCTCCA TCATCAATCA ATCTTCACGA AATCCTACTC
     1321 GGGCGTGTGG CGTAGTTGGT AGCGCGTTCG CTTAGCATGC GAAAGGTCTC CGGTTCGAAT
     1381 CCGGACTCGT CCAGTTTGCG CTTCAAACCA CTTgttttag agctagaaat agcaagttaa
     1441 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttGGCCGGC
     1501 ATGGTCCCAG CCTCCTCGCT GGCGCCGGCT GGGCAACATG CTTCGGCATG GCGAATGGGA
     1561 Ctaactcgag tcagtactga caataaaaag attcttgttt tcaagaactt gtcatttgta
     1621 tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat ttatattttt
     1681 tttcgcctcg acatcatctg cccagatgcg aagttaagtg cgcagaaagt aatatcatgc
     1741 gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta acgccgccat
     1801 ccagtGGATC CTCTAGAGTC GACCTGCAGG CATGCAAGCT TGGCGTAATC ATGGTCATAG
     1861 CTGTTTCCTG TGTGAAATTG TTATCCGCTC ACAATTCCAC ACAACATACG AGCCGGAAGC
     1921 ATAAAGTGTA AAGCCTGGGG TGCCTAATGA GTGAGCTAAC TCACATTAAT TGCGTTGCGC
     1981 TCACTGCCCG CTTTCCAGTC GGGAAACCTG TCGTGCCAGC TGCATTAATG AATCGGCCAA
     2041 CGCGCGGGGA GAGGCGGTTT GCGTATTGGG CGCTCTTCCG CTTCCTCGCT CACTGACTCG
     2101 CTGCGCTCGG TCGTTCGGCT GCGGCGAGCG GTATCAGCTC ACTCAAAGGC GGTAATACGG
     2161 TTATCCACAG AATCAGGGGA TAACGCAGGA AAGAACATGT GAGCAAAAGG CCAGCAAAAG
     2221 GCCAGGAACC GTAAAAAGGC CGCGTTGCTG GCGTTTTTCC ATAGGCTCCG CCCCCCTGAC
     2281 GAGCATCACA AAAATCGACG CTCAAGTCAG AGGTGGCGAA ACCCGACAGG ACTATAAAGA
     2341 TACCAGGCGT TTCCCCCTGG AAGCTCCCTC GTGCGCTCTC CTGTTCCGAC CCTGCCGCTT
     2401 ACCGGATACC TGTCCGCCTT TCTCCCTTCG GGAAGCGTGG CGCTTTCTCA TAGCTCACGC
     2461 TGTAGGTATC TCAGTTCGGT GTAGGTCGTT CGCTCCAAGC TGGGCTGTGT GCACGAACCC
     2521 CCCGTTCAGC CCGACCGCTG CGCCTTATCC GGTAACTATC GTCTTGAGTC CAACCCGGTA
     2581 AGACACGACT TATCGCCACT GGCAGCAGCC ACTGGTAACA GGATTAGCAG AGCGAGGTAT
     2641 GTAGGCGGTG CTACAGAGTT CTTGAAGTGG TGGCCTAACT ACGGCTACAC TAGAAGAACA
     2701 GTATTTGGTA TCTGCGCTCT GCTGAAGCCA GTTACCTTCG GAAAAAGAGT TGGTAGCTCT
     2761 TGATCCGGCA AACAAACCAC CGCTGGTAGC GGTGGTTTTT TTGTTTGCAA GCAGCAGATT
     2821 ACGCGCAGAA AAAAAGGATC TCAAGAAGAT CCTTTGATCT TTTCTACGGG GTCTGACGCT
     2881 CAGTGGAACG AAAACTCACG TTAAGGGATT TTGGTCATGA GATTATCAAA AAGGATCTTC
     2941 ACCTAGATCC TTTTAAATTA AAAATGAAGT TTTAAATCAA TCTAAAGTAT ATATGAGTAA
     3001 ACTTGGTCTG ACAGTTACCA ATGCTTAATC AGTGAGGCAC CTATCTCAGC GATCTGTCTA
     3061 TTTCGTTCAT CCATAGTTGC CTGACTCCCC GTCGTGTAGA TAACTACGAT ACGGGAGGGC
     3121 TTACCATCTG GCCCCAGTGC TGCAATGATA CCGCGAGACC CACGCTCACC GGCTCCAGAT
     3181 TTATCAGCAA TAAACCAGCC AGCCGGAAGG GCCGAGCGCA GAAGTGGTCC TGCAACTTTA
     3241 TCCGCCTCCA TCCAGTCTAT TAATTGTTGC CGGGAAGCTA GAGTAAGTAG TTCGCCAGTT
     3301 AATAGTTTGC GCAACGTTGT TGCCATTGCT ACAGGCATCG TGGTGTCACG CTCGTCGTTT
     3361 GGTATGGCTT CATTCAGCTC CGGTTCCCAA CGATCAAGGC GAGTTACATG ATCCCCCATG
     3421 TTGTGCAAAA AAGCGGTTAG CTCCTTCGGT CCTCCGATCG TTGTCAGAAG TAAGTTGGCC
     3481 GCAGTGTTAT CACTCATGGT TATGGCAGCA CTGCATAATT CTCTTACTGT CATGCCATCC
     3541 GTAAGATGCT TTTCTGTGAC TGGTGAGTAC TCAACCAAGT CATTCTGAGA ATAGTGTATG
     3601 CGGCGACCGA GTTGCTCTTG CCCGGCGTCA ATACGGGATA ATACCGCGCC ACATAGCAGA
     3661 ACTTTAAAAG TGCTCATCAT TGGAAAACGT TCTTCGGGGC GAAAACTCTC AAGGATCTTA
     3721 CCGCTGTTGA GATCCAGTTC GATGTAACCC ACTCGTGCAC CCAACTGATC TTCAGCATCT
     3781 TTTACTTTCA CCAGCGTTTC TGGGTGAGCA AAAACAGGAA GGCAAAATGC CGCAAAAAAG
     3841 GGAATAAGGG CGACACGGAA ATGTTGAATA CTCATACTCT TCCTTTTTCA ATATTATTGA
     3901 AGCATTTATC AGGGTTATTG TCTCATGAGC GGATACATAT TTGAATGTAT TTAGAAAAAT
     3961 AAACAAATAG GGGTTCCGCG CACATTTCCC CGAAAAGTGC CACCTGACGT CTAAGAAACC
     4021 ATTATTATCA TGACATTAAC CTATAAAAAT AGGCGTATCA CGAGGCCCTT TCGTC
//