LOCUS       pDYSCKO_CAN1            6467 bp ds-DNA     circular     13-NOV-2018
DEFINITION  .
KEYWORDS    "marker:Target8FR, AmpR"                                           
FEATURES             Location/Qualifiers
     misc_feature    16..21
                     /label="gcatgc Sph1 site insertion"
     promoter        complement(4385..4653)
                     /label="SNR52 promoter"
     gene            1543..2403
                     /label="AmpR"
     promoter        3492..3509
                     /label="lac promoter"
     promoter        1438..1542
                     /label="AmpR promoter"
     misc_feature    3621..3889
                     /label="SNR52 promoter"
     CDS             1612..2403
                     /label="AmpR"
     rep_origin      69..1411
                     /label="2μ ori"
     rep_origin      2574..3162
                     /label="ori"
     promoter        3585..3603
                     /label="T3 promoter"
     promoter        3486..3491
                     /label="lac promoter"
     misc_feature    3995..4008
                     /label="Sup4 term"
     terminator      4014..4232
                     /label="CYC1 terminator"
     stuffer         complement(4366..4384)
                     /label="DYSCKO stuffer"
     gRNA terminator complement(4267..4280)
                     /label="Sup4 term"
     terminator      4233..4261
                     /label="CYC1 terminator"
     mutation        2262..2262
                     /label="AmpR silent mutation to remove BsaI cut site"
     protein_bind    3524..3540
                     /label="lac operator"
     Terminator      4014..4261
                     /label="CYC1  Term"
     RNA             3618..4657
                     /label="DYSCKO-CAN1 Cassette"
     CDS             1543..1611
                     /label="AmpR"
     primer_bind     3548..3564
                     /label="M13 rev"
     RNA             complement(4285..4365)
                     /label="tracrRNA"
     promoter        3510..3516
                     /label="lac promoter"
     rep_origin      4662..5117
                     /label="f1 ori"
     misc_feature    3890..3909
                     /label="Target_CAN1"
     promoter        complement(6052..6267)
                     /label="URA3 promoter"
     misc_feature    3910..3992
                     /label="tracrRNA"
     protein_bind    401..448
                     /label="FRT"
     CDS             complement(5248..6051)
                     /label="URA3"
     mutation        5380..5380
                     /label="URA3 silent mutation to remove BsaI cut site"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagtatga tccaatatca aaggaaatga tagcattgaa ggatgagact aatccaattg
      121 aggagtggca gcatatagaa cagctaaagg gtagtgctga aggaagcata cgataccccg
      181 catggaatgg gataatatca caggaggtac tagactacct ttcatcctac ataaatagac
      241 gcatataagt acgcatttaa gcataaacac gcactatgcc gttcttctca tgtatatata
      301 tatacaggca acacgcagat ataggtgcga cgtgaacagt gagctgtatg tgcgcagctc
      361 gcgttgcatt ttcggaagcg ctcgttttcg gaaacgcttt gaagttccta ttccgaagtt
      421 cctattctct agaaagtata ggaacttcag agcgcttttg aaaaccaaaa gcgctctgaa
      481 gacgcacttt caaaaaacca aaaacgcacc ggactgtaac gagctactaa aatattgcga
      541 ataccgcttc cacaaacatt gctcaaaagt atctctttgc tatatatctc tgtgctatat
      601 ccctatataa cctacccatc cacctttcgc tccttgaact tgcatctaaa ctcgacctct
      661 acatttttta tgtttatctc tagtattact ctttagacaa aaaaattgta gtaagaacta
      721 ttcatagagt gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg tagtatatag
      781 agacaaaata gaagaaaccg ttcataattt tctgaccaat gaagaatcat caacgctatc
      841 actttctgtt cacaaagtat gcgcaatcca catcggtata gaatataatc ggggatgcct
      901 ttatcttgaa aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg gaagtggagt
      961 caggcttttt ttatggaaga gaaaatagac accaaagtag ccttcttcta accttaacgg
     1021 acctacagtg caaaaagtta tcaagagact gcattataga gcgcacaaag gagaaaaaaa
     1081 gtaatctaag atgctttgtt agaaaaatag cgctctcggg atgcattttt gtagaacaaa
     1141 aaagaagtat agattctttg ttggtaaaat agcgctctcg cgttgcattt ctgttctgta
     1201 aaaatgcagc tcagattctt tgtttgaaaa attagcgctc tcgcgttgca tttttgtttt
     1261 acaaaaatga agcacagatt cttcgttggt aaaatagcgc tttcgcgttg catttctgtt
     1321 ctgtaaaaat gcagctcaga ttctttgttt gaaaaattag cgctctcgcg ttgcattttt
     1381 gttctacaaa atgaagcaca gatgcttcgt tcaggtggca cttttcgggg aaatgtgcgc
     1441 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1501 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1561 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1621 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1681 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1741 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     1801 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1861 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1921 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     1981 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2041 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     2101 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     2161 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     2221 tggtttattg ctgataaatc tggagccggt gagcgtgggt cCcgcggtat cattgcagca
     2281 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     2341 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     2401 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     2461 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2521 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2581 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2641 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2701 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     2761 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2821 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2881 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2941 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3001 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3061 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3241 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3301 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     3361 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     3421 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt acctcactca ttaggcaccc
     3481 caggctttac actttatgct tccggctcct atgttgtgtg gaattgtgag cggataacaa
     3541 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
     3601 agggaacaaa agctggagct tctttgaaaa gataatgtat gattatgctt tcactcatat
     3661 ttatacagaa acttgatgtt ttctttcgag tatatacaag gtgattacat gtacgtttga
     3721 agtacaactc tagattttgt agtgccctct tgggctagcg gtaaaggtgc gcattttttc
     3781 acaccctaca atgttctgtt caaaagattt tggtcaaacg ctgtagaagt gaaagttggt
     3841 gcgcatgttt cggcgttcga aacttctccg cagtgaaaga taaatgatcA TAACGGAATC
     3901 CAACTGGGCG TTTTAGAGCT AGAAATAGCA AGTTAAAATA AGGCTAGTCC GTTATCAACT
     3961 TGAAAAAGTG GCACCGAGTC GGTGGTGCTT TTTTTGTTTT TTATGTCTtc gagtcatgta
     4021 attagttatg tcacgcttac Gttcacgccc tccccccaca tccgctctaa ccgaaaagga
     4081 aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt
     4141 aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg
     4201 taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg
     4261 cctcgaAGAC ATAAAAAACA AAAAAAGCAC CACCGACTCG GTGCCACTTT TTCAAGTTGA
     4321 TAACGGACTA GCCTTATTTT AACTTGCTAT TTCTAGCTCT AAAACTGAGA CCAGATCTGG
     4381 TCTCgatcat ttatctttca ctgcggagaa gtttcgaacg ccgaaacatg cgcaccaact
     4441 ttcacttcta cagcgtttga ccaaaatctt ttgaacagaa cattgtaggg tgtgaaaaaa
     4501 tgcgcacctt taccgctagc ccaagagggc actacaaaat ctagagttgt acttcaaacg
     4561 tacatgtaat caccttgtat atactcgaaa gaaaacatca agtttctgta taaatatgag
     4621 tgaaagcata atcatacatt atcttttcaa agaatggcgc gacgcgccct gtagcggcgc
     4681 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4741 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4801 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4861 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4921 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4981 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     5041 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     5101 attaacgttt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     5161 caccgcatag ggtaataact gatataatta aattgaagct ctaatttgtg agtttagtat
     5221 acatgcattt acttataata cagtttttta gttttgctgg ccgcatcttc tcaaatatgc
     5281 ttcccagcct gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa
     5341 tagtcctctt ccaacaataa taatgtcaga tcctgtagaA accacatcat ccacggttct
     5401 atactgttga cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg tcataatcaa
     5461 ccaatcgtaa ccttcatctc ttccacccat gtctctttga gcaataaagc cgataacaaa
     5521 atctttgtcg ctcttcgcaa tgtcaacagt acccttagta tattctccag tagataggga
     5581 gcccttgcat gacaattctg ctaacatcaa aaggcctcta ggttcctttg ttacttcttc
     5641 tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg cattcgtaat
     5701 gtctgcccat tctgctattc tgtatacacc cgcagagtac tgcaatttga ctgtattacc
     5761 aatgtcagca aattttctgt cttcgaagag taaaaaattg tacttggcgg ataatgcctt
     5821 tagcggctta actgtgccct ccatggaaaa atcagtcaag atatccacat gtgtttttag
     5881 taaacaaatt ttgggaccta atgcttcaac taactccagt aattccttgg tggtacgaac
     5941 atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg atattaaata gcttggcagc
     6001 aacaggacta ggatgagtag cagcacgttc cttatatgta gctttcgaca tgatttatct
     6061 tcgtttcctg caggtttttg ttctgtgcag ttgggttaag aatactgggc aatttcatgt
     6121 ttcttcaaca ctacatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc
     6181 ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa Ggaaaccgaa atcaaaaaaa
     6241 agaataaaaa aaaaatgatg aattgaattg aaaagctgtg gtatggtgca ctctcagtac
     6301 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     6361 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     6421 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcga
//