LOCUS       pCK071                  7638 bp ds-DNA     circular     09-MAY-2025
DEFINITION  .
KEYWORDS    "Parental Vector" "Origin" "Resistance" "Construction Method"
            "Promoter" "Terminator" "gRNA" "E.coli plasmids" "pCD"
FEATURES             Location/Qualifiers
     terminator      3917..3984
                     /label="sglTerm"
                     /ApEinfo_revcolor="#800040"
                     /ApEinfo_fwdcolor="#800040"
     terminator      3917..4045
                     /label="dblTerm"
                     /ApEinfo_revcolor="#800080"
                     /ApEinfo_fwdcolor="#800080"
     promoter        4092..4126
                     /label="J23107 promoter"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    4127..4153
                     /label="Bujard RBS (strong)"
                     /ApEinfo_revcolor="#db7aff"
                     /ApEinfo_fwdcolor="#db7aff"
     RBS             4136..4147
                     /label="RBS (strong)"
                     /ApEinfo_revcolor="#996633"
                     /ApEinfo_fwdcolor="#996633"
     CDS             4154..5119
                     /label="Translation 4154-5119"
                     /translation="MGPMAEYKDNLLGEANSFLEVLEQVSHLAPLDKPVLIIGERGTGKELIASRLHYLSSRWQGPFISLNCAALNENLLDSELFGHEAGAFTGAQKRHPGRFERADGGTLFLDELATAPMMVQEKLLRVIEYGELERVGGSQPLQVNVRLVCATNADLPAMVNEGTFRADLLDRLAFDVVQLPPLRERESDIMLMAEYFAIQMCREIKLPLFPGFTERARETLLNYRWPGNIRELKNVVERSVYRHGTSDYPLDDIIIDPFKRRPPEDAIAVSETTSLPTLPLDLREFQMQQEKELLQLSLQMNARTRRRERRAEKQAQWKAAN*"
     misc_feature    4163..5050
                     /label="PspF-dHTH"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    5051..5119
                     /label="lambdaN22Plus"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     terminator      5130..5163
                     /label="BBa_B1002 terminator"
                     /ApEinfo_revcolor="#af14ff"
                     /ApEinfo_fwdcolor="#af14ff"
     misc_feature    5164..5881
                     /label="p15A origin"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    5317..5879
                     /label="p15A origin"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    5972..6077
                     /label="Phage lambda T0 terminator"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    complement(5987..6850)
                     /label="CmR(EcoRI-KO) "
                     /ApEinfo_revcolor="#120eff"
                     /ApEinfo_fwdcolor="#120eff"
     misc_feature    complement(6096..6752)
                     /label="CmR (EcoRI-KO)"
                     /ApEinfo_revcolor="#000bff"
                     /ApEinfo_fwdcolor="#000bff"
     promoter        complement(6819..6850)
                     /label="pCat promoter (weak)"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     promoter        6890..7432
                     /label="endogenous promoter"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     misc_feature    7433..3898
                     /label="dCas9"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
ORIGIN
        1 TAGAAGGTAT ACACGTCGGA AGAATCGTAT TTGTTATCTA CAGGAGATTT TTTCAAATGA
       61 GATGGCGAAA GTAGATGATA GTTTCTTTCA TCGACTTGAA GAGTCTTTTT TGGTGGAAGA
      121 AGACAAGAAG CATGAACGTC ATCCTATTTT TGGAAATATA GTAGATGAAG TTGCTTATCA
      181 TGAGAAATAT CCAACTATCT ATCATCTGCG AAAAAAATTG GTAGATTCTA CTGATAAAGC
      241 GGATTTGCGC TTAATCTATT TGGCCTTAGC GCATATGATT AAGTTTCGTG GTCATTTTTT
      301 GATTGAGGGA GATTTAAATC CTGATAATAG TGATGTGGAC AAACTATTTA TCCAGTTGGT
      361 ACAAACCTAC AATCAATTAT TTGAAGAAAA CCCTATTAAC GCAAGTGGAG TAGATGCTAA
      421 AGCGATTCTT TCTGCACGAT TGAGTAAATC AAGACGATTA GAAAATCTCA TTGCTCAGCT
      481 CCCCGGTGAG AAGAAAAATG GCTTATTTGG GAATCTCATT GCTTTGTCAT TGGGTTTGAC
      541 CCCTAATTTT AAATCAAATT TTGATTTGGC AGAAGATGCT AAATTACAGC TTTCAAAAGA
      601 TACTTACGAT GATGATTTAG ATAATTTATT GGCGCAAATT GGAGATCAAT ATGCTGATTT
      661 GTTTTTGGCA GCTAAGAATT TATCAGATGC TATTTTACTT TCAGATATCC TAAGAGTAAA
      721 TACTGAAATA ACTAAGGCTC CCCTATCAGC TTCAATGATT AAACGCTACG ATGAACATCA
      781 TCAAGACTTG ACTCTTTTAA AAGCTTTAGT TCGACAACAA CTTCCAGAAA AGTATAAAGA
      841 AATCTTTTTT GATCAATCAA AAAACGGATA TGCAGGTTAT ATTGATGGGG GAGCTAGCCA
      901 AGAAGAATTT TATAAATTTA TCAAACCAAT TTTAGAAAAA ATGGATGGTA CTGAGGAATT
      961 ATTGGTGAAA CTAAATCGTG AAGATTTGCT GCGCAAGCAA CGGACCTTTG ACAACGGCTC
     1021 TATTCCCCAT CAAATTCACT TGGGTGAGCT GCATGCTATT TTGAGAAGAC AAGAAGACTT
     1081 TTATCCATTT TTAAAAGACA ATCGTGAGAA GATTGAAAAA ATCTTGACTT TTCGAATTCC
     1141 TTATTATGTT GGTCCATTGG CGCGTGGCAA TAGTCGTTTT GCATGGATGA CTCGGAAGTC
     1201 TGAAGAAACA ATTACCCCAT GGAATTTTGA AGAAGTTGTC GATAAAGGTG CTTCAGCTCA
     1261 ATCATTTATT GAACGCATGA CAAACTTTGA TAAAAATCTT CCAAATGAAA AAGTACTACC
     1321 AAAACATAGT TTGCTTTATG AGTATTTTAC GGTTTATAAC GAATTGACAA AGGTCAAATA
     1381 TGTTACTGAA GGAATGCGAA AACCAGCATT TCTTTCAGGT GAACAGAAGA AAGCCATTGT
     1441 TGATTTACTC TTCAAAACAA ATCGAAAAGT AACCGTTAAG CAATTAAAAG AAGATTATTT
     1501 CAAAAAAATA GAATGTTTTG ATAGTGTTGA AATTTCAGGA GTTGAAGATA GATTTAATGC
     1561 TTCATTAGGT ACCTACCATG ATTTGCTAAA AATTATTAAA GATAAAGATT TTTTGGATAA
     1621 TGAAGAAAAT GAAGATATCT TAGAGGATAT TGTTTTAACA TTGACCTTAT TTGAAGATAG
     1681 GGAGATGATT GAGGAAAGAC TTAAAACATA TGCTCACCTC TTTGATGATA AGGTGATGAA
     1741 ACAGCTTAAA CGTCGCCGTT ATACTGGTTG GGGACGTTTG TCTCGAAAAT TGATTAATGG
     1801 TATTAGGGAT AAGCAATCTG GCAAAACAAT ATTAGATTTT TTGAAATCAG ATGGTTTTGC
     1861 CAATCGCAAT TTTATGCAGC TGATCCATGA TGATAGTTTG ACATTTAAAG AAGACATTCA
     1921 AAAAGCACAA GTGTCTGGAC AAGGCGATAG TTTACATGAA CATATTGCAA ATTTAGCTGG
     1981 TAGCCCTGCT ATTAAAAAAG GTATTTTACA GACTGTAAAA GTTGTTGATG AATTGGTCAA
     2041 AGTAATGGGG CGGCATAAGC CAGAAAATAT CGTTATTGAA ATGGCACGTG AAAATCAGAC
     2101 AACTCAAAAG GGCCAGAAAA ATTCGCGAGA GCGTATGAAA CGAATCGAAG AAGGTATCAA
     2161 AGAATTAGGA AGTCAGATTC TTAAAGAGCA TCCTGTTGAA AATACTCAAT TGCAAAATGA
     2221 AAAGCTCTAT CTCTATTATC TCCAAAATGG AAGAGACATG TATGTGGACC AAGAATTAGA
     2281 TATTAATCGT TTAAGTGATT ATGATGTCGA TgcCATTGTT CCACAAAGTT TCCTTAAAGA
     2341 CGATTCAATA GACAATAAGG TCTTAACGCG TTCTGATAAA AATCGTGGTA AATCGGATAA
     2401 CGTTCCAAGT GAAGAAGTAG TCAAAAAGAT GAAAAACTAT TGGAGACAAC TTCTAAACGC
     2461 CAAGTTAATC ACTCAACGTA AGTTTGATAA TTTAACGAAA GCTGAACGTG GAGGTTTGAG
     2521 TGAACTTGAT AAAGCTGGTT TTATCAAACG CCAATTGGTT GAAACTCGCC AAATCACTAA
     2581 GCATGTGGCA CAAATTTTGG ATAGTCGCAT GAATACTAAA TACGATGAAA ATGATAAACT
     2641 TATTCGAGAG GTTAAAGTGA TTACCTTAAA ATCTAAATTA GTTTCTGACT TCCGAAAAGA
     2701 TTTCCAATTC TATAAAGTAC GTGAGATTAA CAATTACCAT CATGCCCATG ATGCGTATCT
     2761 AAATGCCGTC GTTGGAACTG CTTTGATTAA GAAATATCCA AAACTTGAAT CGGAGTTTGT
     2821 CTATGGTGAT TATAAAGTTT ATGATGTTCG TAAAATGATT GCTAAGTCTG AGCAAGAAAT
     2881 AGGCAAAGCA ACCGCAAAAT ATTTCTTTTA CTCTAATATC ATGAACTTCT TCAAAACAGA
     2941 AATTACACTT GCAAATGGAG AGATTCGCAA ACGCCCTCTA ATCGAAACTA ATGGGGAAAC
     3001 TGGAGAAATT GTCTGGGATA AAGGGCGAGA TTTTGCCACA GTGCGCAAAG TATTGTCCAT
     3061 GCCCCAAGTC AATATTGTCA AGAAAACAGA AGTACAGACA GGCGGATTCT CCAAGGAGTC
     3121 AATTTTACCA AAAAGAAATT CGGACAAGCT TATTGCTCGT AAAAAAGACT GGGATCCAAA
     3181 AAAATATGGT GGTTTTGATA GTCCAACGGT AGCTTATTCA GTCCTAGTGG TTGCTAAGGT
     3241 GGAAAAAGGG AAATCGAAGA AGTTAAAATC CGTTAAAGAG TTACTAGGGA TCACAATTAT
     3301 GGAAAGAAGT TCCTTTGAAA AAAATCCGAT TGACTTTTTA GAAGCTAAAG GATATAAGGA
     3361 AGTTAAAAAA GACTTAATCA TTAAACTACC TAAATATAGT CTTTTTGAGT TAGAAAACGG
     3421 TCGTAAACGG ATGCTGGCTA GTGCCGGAGA ATTACAAAAA GGAAATGAGC TGGCTCTGCC
     3481 AAGCAAATAT GTGAATTTTT TATATTTAGC TAGTCATTAT GAAAAGTTGA AGGGTAGTCC
     3541 AGAAGATAAC GAACAAAAAC AATTGTTTGT GGAGCAGCAT AAGCATTATT TAGATGAGAT
     3601 TATTGAGCAA ATCAGTGAAT TTTCTAAGCG TGTTATTTTA GCAGATGCCA ATTTAGATAA
     3661 AGTTCTTAGT GCATATAACA AACATAGAGA CAAACCAATA CGTGAACAAG CAGAAAATAT
     3721 TATTCATTTA TTTACGTTGA CGAATCTTGG AGCTCCCGCT GCTTTTAAAT ATTTTGATAC
     3781 AACAATTGAT CGTAAACGAT ATACGTCTAC AAAAGAAGTT TTAGATGCCA CTCTTATCCA
     3841 TCAATCCATC ACTGGTCTTT ATGAAACACG CATTGATTTG AGTCAGCTAG GAGGTGACTA
     3901 ACtcgagtaa ggatctccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     3961 tttcgtttta tctgttgttt gtcggtgaac gctctctact agagtcacac tggctcacct
     4021 tcgggtgggc ctttctgcgt ttataCCTAG agcgcccgga agagagtcaa ttcagggtgg
     4081 tgaatctgca gtttacggct agctcagccc taggtattat gctagcGAAT TCATTAAAGA
     4141 GGAGAAAGGT ACCatggggc ccatggcaga atacaaagat aatttacttg gtgaggcgaa
     4201 cagctttctc gaagtgctgg aacaggtttc gcatctcgca ccgctggaca aaccggtgct
     4261 catcatcggc gaacgcggca ccggtaaaga gctgattgcc agccgcctgc attatctctc
     4321 ctcccgttgg caagggccgt ttatttccct taactgcgcg gcgttaaatg aaaatctgct
     4381 ggattccgaa ctgtttggtc acgaagcggg ggcgtttacc ggtgcgcaaa aacgtcatcc
     4441 agggagattt gaacgtgccg acggcggtac gctatttctt gatgaactcg ctacggcacc
     4501 gatgatggtg caggagaaat tattgcgcgt gattgagtac ggtgaactgg agcgcgttgg
     4561 cggcagccaa ccattgcagg tgaatgtgcg gttggtatgc gcgacgaatg ccgatctccc
     4621 ggcgatggtc aatgaaggca cttttcgcgc tgacctgctc gaccgactgg cttttgatgt
     4681 tgtacaactg ccaccactgc gcgagcgcga aagcgacata atgttgatgg cagaatactt
     4741 tgccatccag atgtgtcggg aaatcaagct gcctctgttc ccggggttta cggagcgcgc
     4801 cagagaaaca ttgctgaatt atcgttggcc gggaaatatt cgtgaattga aaaacgtggt
     4861 ggaacgttca gtgtatcgcc acggcaccag cgattatccg cttgatgaca tcattattga
     4921 tccctttaaa cggcgtccgc ctgaagacgc tatcgccgtt tcagaaacca cctcgcttcc
     4981 aacactgccg ctggatttac gtgagtttca gatgcagcag gaaaaagagt tgctgcaact
     5041 cagtttgcaa ATGAATGCAC GCACACGCCG CCGCGAACGT CGCGCAGAGA AACAGGCTCA
     5101 ATGGAAAGCA GCAAATTAAG CGGCCGCcac gcaaaaaacc ccgcttcggc ggggtttttt
     5161 cgccctaggg atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact
     5221 gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat
     5281 acttaacagg gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct
     5341 gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa
     5401 agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg
     5461 tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag
     5521 ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga
     5581 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc
     5641 accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg
     5701 gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc cagttacctc
     5761 ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt
     5821 cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat catcttatta
     5881 atcagataaa atatttctag atttcagtgc aatttatctc ttcaaatgta gcacctgaag
     5941 tcagccccat acgatataag ttgttactag tgcttggatt ctcaccaata aaaaacgccc
     6001 ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta
     6061 tcaacaggag tccaagcgag ctcgatatca aattacgccc cgccctgcca ctcatcgcag
     6121 tactgttgta attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga
     6181 acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg
     6241 aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc
     6301 acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc
     6361 aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg
     6421 tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa
     6481 caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat acgaaattcc
     6541 ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta
     6601 tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta
     6661 cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca
     6721 acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc
     6781 gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct
     6841 cttacgtgcc gatcaacgtc tcattttcgc cagatatcga cgtcttaagT TACGAAATCA
     6901 TCCTGTGGAG CTTAGTAGGT TTAGCAAGAT GGCAGCGCCT AAATGTAGAA TGATAAAAGG
     6961 ATTAAGAGAT TAATTTCCCT AAAAATGATA AAACAAGCGT TTTGAAAGCG CTTGTTTTTT
     7021 TGGTTTGCAG TCAGAGTAGA ATAGAAGTAT CAAAAAAAGC ACCGACTCGG TGCCACTTTT
     7081 TCAAGTTGAT AACGGACTAG CCTTATTTTA ACTTGCTATG CTGTTTTGAA TGGTTCCAAC
     7141 AAGATTATTT TATAACTTTT ATAACAAATA ATCAAGGAGA AATTCAAAGA AATTTATCAG
     7201 CCATAAAACA ATACTTAATA CTATAGAATG ATAACAAAAT AAACTACTTT TTAAAAGAAT
     7261 TTTGTGTTAT AATCTATTTA TTATTAAGTA TTGGGTAATA TTTTTTGAAG AGATATTTTG
     7321 AAAAAGAAAA ATTAAAGCAT ATTAAACTAA TTTCGGAGGT CATTAAAACT ATTATTGAAA
     7381 TCATCAAACT CATTATGGAT TTAATTTAAA CTTTTTATTT TAGGAGGCAA AAATGGATAA
     7441 GAAATACTCA ATAGGCTTAG cTATCGGCAC AAATAGCGTC GGATGGGCGG TGATCACTGA
     7501 TGAATATAAG GTTCCGTCTA AAAAGTTCAA GGTTCTGGGA AATACAGACC GCCACAGTAT
     7561 CAAAAAAAAT CTTATAGGGG CTCTTTTATT TGACAGTGGA GAGACAGCGG AAGCGACTCG
     7621 TCTCAAACGG ACAGCTCG
//