LOCUS       pCas9-NT                7271 bp ds-DNA     circular     26-FEB-2025
DEFINITION  .
KEYWORDS    "creator:Enrico Orsi"
COMMENT     Imported using the Genbank importer. File name:
            1_psibr-cas9_constitutive_nt_ev.gb Imported using the Genbank
            importer. File name: psibr-cas9_constitutive_nt_ev.gb Imported
            using the Genbank importer. File name: psibr-cas9_int3_nt.gb
FEATURES             Location/Qualifiers
     misc_feature    complement(318..1089)
                     /label="Ori pBBR1"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
                     /note="label: Ori pBBR1"
     misc_feature    1334..1363
                     /label="lcaUV5 promoter"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /note="label: lcaUV5 promoter"
     misc_feature    1432..5538
                     /label="Cas9"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     CDS             1435..5535
                     /label="CDS"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD"
     misc_feature    5586..5607
                     /label="B1002 terminator"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
                     /note="label: B1002 terminator"
     promoter        5614..5644
                     /label="lacUV5 promoter"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /note="label: lacUV5 promoter"
     misc_feature    5645..5679
                     /label="NT sequence"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_RNA        5680..5755
                     /label="gRNA scaffold"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
                     /note="note: guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system"
     terminator      5911..5932
                     /label="B1002 terminator"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
                     /note="label: B1002 terminator"
     misc_feature    6040..6855
                     /label="Kan Neo"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
                     /note="label: Kan Neo"
     misc_feature    complement(6929..317)
                     /label="'rep"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /note="label: 'rep"
ORIGIN
        1 TCATTGACCA CGTAGGCCGA CACGGTGCCG GGGCCGTTGA GCTTCACGAC GGAGATCCAG
       61 CGCTCGGCCA CCAAGTCCTT GACTGCGTAT TGGACCGTCC GCAAAGAACG TCCGATGAGC
      121 TTGGAAAGTG TTTTCTGGCT GACCACCACG GCGTTCTGGT GGCCCATCTG CGCCACGAGG
      181 TGATGCAGCA GCATTGCCGC CGTGGGTTTC CTCGCAATAA GCCCGGCCCA CGCCTCATGC
      241 GCTTTGCGTT CCGTTTGCAC CCAGTGACCG GGCTTGTTCT TGGCTTGAAT GCCGATTTCT
      301 CTGGACTGCG TGGCCATGCT TATCTCCATG CGGTAGGGGT GCCGCACGGT TGCGGCACCA
      361 TGCGCAATCA GCTGCAACTT TTCGGCAGCG CGACAACAAT TATGCGTTGC GTAAAAGTGG
      421 CAGTCAATTA CAGATTTTCT TTAACCTACG CAATGAGCTA TTGCGGGGGG TGCCGCAATG
      481 AGCTGTTGCG TACCCCCCTT TTTTAAGTTG TTGATTTTTA AGTCTTTCGC ATTTCGCCCT
      541 ATATCTAGTT CTTTGGTGCC CAAAGAAGGG CACCCCTGCG GGGTTCCCCC ACGCCTTCGG
      601 CGCGGCTCCC CCTCCGGCAA AAAGTGGCCC CTCCGGGGCT TGTTGATCGA CTGCGCGGCC
      661 TTCGGCCTTG CCCAAGGTGG CGCTGCCCCC TTGGAACCCC CGCACTCGCC GCCGTGAGGC
      721 TCGGGGGGCA GGCGGGCGGG CTTCGCCCTT CGACTGCCCC CACTCGCATA GGCTTGGGTC
      781 GTTCCAGGCG CGTCAAGGCC AAGCCGCTGC GCGGTCGCTG CGCGAGCCTT GACCCGCCTT
      841 CCACTTGGTG TCCAACCGGC AAGCGAAGCG CGCAGGCCGC AGGCCGGAGG CTTTTCCCCA
      901 GAGAAAATTA AAAAAATTGA TGGGGCAAGG CCGCAGGCCG CGCAGTTGGA GCCGGTGGGT
      961 ATGTGGTCGA AGGCTGGGTA GCCGGTGGGC AATCCCTGTG GTCAAGCTCG TGGGCAGGCG
     1021 CAGCCTGTCC ATCAGCTTGT CCAGCAGGGT TGTCCACGGG CCGAGCGAAG CGAGCCAGCC
     1081 GGTGGCCGCT CGCGGCCATC GTCCACATAT CCACGGGCTG GCAAGGGAGC GCAGCGACCG
     1141 CGCAGGGCGA AGCCCGGAGA GCAAGCCCGT AGGGGGGGCG CGCCCAGCTG TCTAGGGCGG
     1201 CGGATTTGTC CTACTCAGGA GAGCGTTCAC CGACAAACAA CAGATAAAAC GAAAGGCCCA
     1261 GTCTTTCGAC TGAGCCTTTC GTTTTATTTG ATGCCTTTAA TTAAAGCGGA TAACAATTTC
     1321 ACACAGGAGG CCGTTTACAC TTTATGCTTC CGGCTCGTAT AATGTGTGGG GAAACCACAA
     1381 CGGTTTCCCT CTAGAAATAA TTTTGTTTAA CTATAAGAAG GAGATATACA TATGGACAAA
     1441 AAGTACTCGA TTGGGCTCGA CATCGGGACC AACTCCGTTG GGTGGGCTGT TATCACCGAC
     1501 GAGTACAAAG TCCCTTCGAA GAAATTTAAA GTCTTGGGGA ACACCGATCG TCATAGCATC
     1561 AAGAAGAACC TTATTGGAGC GCTTCTCTTC GATAGCGGGG AAACCGCTGA GGCTACCCGG
     1621 TTGAAGCGAA CCGCGCGGAG GAGGTACACC CGGCGAAAAA ACCGGATCTG CTACCTTCAG
     1681 GAAATCTTCT CGAACGAAAT GGCTAAGGTT GACGACAGCT TTTTCCACCG TCTTGAGGAA
     1741 TCGTTCCTTG TTGAGGAGGA TAAAAAACAC GAGCGGCACC CCATCTTCGG GAACATTGTT
     1801 GACGAGGTCG CGTACCACGA AAAGTACCCC ACCATCTACC ACTTGCGTAA GAAGCTTGTT
     1861 GACTCGACCG ACAAGGCTGA CCTTCGTCTC ATCTACCTTG CACTCGCTCA CATGATCAAA
     1921 TTCCGGGGGC ACTTCCTTAT CGAAGGGGAC CTCAACCCCG ACAACAGCGA CGTTGATAAG
     1981 CTTTTCATCC AGCTTGTTCA GACGTACAAC CAGCTCTTCG AGGAGAACCC CATCAACGCT
     2041 AGCGGGGTTG ACGCGAAGGC TATCCTTTCG GCTCGTCTTA GCAAGTCGAG GCGTCTCGAG
     2101 AACTTGATCG CGCAGTTGCC TGGGGAAAAA AAGAACGGGC TCTTCGGAAA CTTGATCGCG
     2161 CTTTCGCTTG GGCTTACGCC CAACTTCAAG TCGAACTTCG ACCTTGCTGA GGACGCGAAG
     2221 CTCCAGCTTT CGAAGGACAC CTACGACGAC GACCTCGACA ACCTCCTTGC TCAGATCGGG
     2281 GACCAGTACG CGGACCTTTT CCTTGCTGCG AAAAACCTCT CGGACGCGAT CCTCCTTTCG
     2341 GACATCCTTA GGGTTAACAC CGAGATTACC AAAGCGCCTC TTTCGGCGTC GATGATCAAG
     2401 CGTTACGACG AGCACCACCA GGATCTTACC CTTCTCAAGG CGCTCGTCCG TCAGCAGCTT
     2461 CCCGAGAAAT ACAAGGAGAT CTTCTTCGAC CAGTCGAAGA ACGGGTACGC TGGGTACATC
     2521 GACGGAGGGG CGTCCCAGGA GGAGTTCTAC AAGTTCATCA AGCCCATCCT CGAGAAGATG
     2581 GACGGGACCG AAGAGCTCCT TGTTAAGCTT AACCGGGAGG ACCTTTTGCG TAAACAGCGA
     2641 ACGTTCGATA ACGGGTCGAT CCCTCACCAG ATCCATCTTG GGGAATTGCA CGCGATCCTT
     2701 AGGAGGCAGG AGGATTTCTA CCCCTTCCTC AAGGATAACC GGGAAAAAAT CGAGAAGATC
     2761 CTTACCTTCC GTATCCCCTA CTACGTCGGG CCCCTTGCTC GGGGGAACAG CCGGTTCGCT
     2821 TGGATGACCC GAAAATCGGA GGAGACCATC ACGCCCTGGA ACTTCGAGGA GGTCGTTGAC
     2881 AAGGGGGCGT CGGCGCAGTC GTTCATCGAG CGTATGACCA ACTTCGACAA GAACCTTCCC
     2941 AACGAGAAGG TTCTTCCCAA GCACAGCCTT CTTTACGAAT ACTTCACGGT CTACAACGAG
     3001 CTTACCAAAG TTAAGTACGT CACCGAGGGG ATGCGTAAGC CCGCTTTCCT TTCGGGGGAG
     3061 CAGAAAAAGG CAATCGTCGA CCTCTTGTTT AAGACCAACC GTAAGGTTAC GGTCAAACAG
     3121 CTCAAGGAGG ACTACTTTAA GAAGATTGAG TGCTTCGACA GCGTCGAGAT CTCGGGGGTC
     3181 GAGGACAGGT TCAACGCGTC GCTCGGGACG TACCACGACC TTCTTAAGAT CATCAAGGAC
     3241 AAGGACTTCC TTGACAACGA GGAGAACGAG GACATCCTCG AAGACATCGT CCTCACCCTT
     3301 ACGCTCTTCG AGGACAGGGA AATGATCGAA GAGAGGCTTA AGACCTACGC GCATTTGTTC
     3361 GACGACAAAG TTATGAAGCA GCTTAAGCGG CGTCGGTACA CCGGGTGGGG GCGGCTTTCG
     3421 CGTAAGCTTA TCAACGGGAT CAGGGACAAA CAGTCGGGGA AGACCATTCT CGACTTCCTT
     3481 AAGTCGGACG GGTTCGCAAA CCGTAACTTC ATGCAGTTGA TCCACGACGA CAGCCTTACC
     3541 TTCAAGGAGG ATATCCAGAA GGCTCAGGTT TCGGGGCAGG GGGACAGCCT CCACGAGCAC
     3601 ATCGCTAACC TCGCGGGGTC CCCCGCGATC AAGAAGGGGA TCCTCCAGAC CGTTAAGGTC
     3661 GTCGACGAGC TTGTTAAGGT TATGGGACGA CACAAACCCG AGAACATCGT CATCGAGATG
     3721 GCTCGGGAGA ACCAGACCAC CCAGAAAGGG CAGAAGAACT CCCGTGAACG GATGAAGCGT
     3781 ATCGAGGAGG GGATCAAGGA GCTCGGGAGC CAGATCCTTA AGGAACACCC CGTCGAGAAC
     3841 ACCCAGCTTC AGAACGAGAA ATTGTACTTG TACTACTTGC AGAACGGGAG GGATATGTAC
     3901 GTTGATCAGG AGCTCGACAT CAACCGGCTC AGCGACTACG ACGTTGACCA TATCGTCCCC
     3961 CAGAGCTTTC TTAAGGATGA CTCGATTGAT AACAAAGTTC TCACGCGGTC GGACAAGAAC
     4021 CGGGGGAAGT CCGACAACGT CCCCAGCGAG GAGGTTGTTA AGAAAATGAA GAACTACTGG
     4081 AGGCAGCTTC TTAACGCAAA ACTCATCACC CAGCGGAAAT TCGACAACCT CACGAAGGCG
     4141 GAGCGGGGGG GGCTTAGCGA GCTTGACAAG GCGGGGTTCA TCAAGCGTCA GCTTGTCGAG
     4201 ACCCGTCAGA TCACCAAACA CGTTGCTCAG ATCCTTGACA GCCGTATGAA CACCAAGTAC
     4261 GACGAGAACG ACAAGCTTAT CCGTGAAGTC AAGGTTATCA CGCTCAAGTC GAAGCTCGTC
     4321 TCGGATTTTC GTAAGGACTT TCAGTTTTAC AAGGTTCGGG AAATCAACAA CTACCACCAC
     4381 GCACACGACG CTTACCTTAA CGCAGTTGTC GGGACCGCGC TTATCAAAAA GTACCCCAAG
     4441 CTTGAGTCCG AATTCGTTTA CGGGGACTAC AAGGTCTACG ACGTCCGGAA GATGATCGCG
     4501 AAATCGGAAC AGGAGATTGG GAAGGCTACG GCTAAGTACT TTTTCTACTC GAACATCATG
     4561 AACTTTTTTA AGACCGAGAT CACCCTTGCT AACGGGGAAA TCCGTAAGCG TCCCCTTATC
     4621 GAGACCAACG GAGAGACCGG GGAGATCGTT TGGGACAAGG GACGTGACTT CGCAACCGTT
     4681 CGTAAGGTTC TTTCCATGCC TCAGGTTAAC ATCGTTAAAA AGACCGAGGT TCAGACCGGG
     4741 GGGTTTTCCA AAGAATCGAT CCTCCCCAAG AGGAACTCCG ATAAACTTAT CGCGCGGAAG
     4801 AAGGATTGGG ACCCCAAGAA GTACGGGGGG TTCGACAGCC CCACGGTTGC GTACTCGGTT
     4861 CTTGTTGTCG CGAAAGTTGA GAAGGGAAAG TCCAAAAAAC TCAAGTCCGT CAAGGAACTC
     4921 CTTGGAATCA CCATCATGGA GAGGAGCTCC TTCGAGAAGA ACCCTATCGA TTTCCTCGAG
     4981 GCGAAGGGGT ACAAAGAGGT CAAGAAGGAT CTCATCATCA AGCTTCCCAA GTACAGCCTT
     5041 TTCGAACTCG AGAACGGGCG GAAGCGAATG TTGGCGAGCG CAGGGGAGCT CCAGAAGGGG
     5101 AACGAATTGG CGTTGCCCTC CAAGTACGTT AACTTCCTCT ACCTCGCGAG CCACTACGAG
     5161 AAACTTAAAG GGAGCCCCGA GGACAACGAG CAGAAGCAGC TTTTCGTTGA ACAGCACAAA
     5221 CACTACCTCG ACGAAATCAT CGAACAGATC AGCGAGTTCT CGAAACGGGT CATCCTCGCT
     5281 GACGCAAACC TCGACAAGGT CCTTAGCGCT TACAACAAGC ACAGGGATAA GCCCATTCGG
     5341 GAGCAGGCTG AGAACATCAT CCACCTCTTC ACGCTTACGA ACCTTGGGGC GCCTGCGGCG
     5401 TTCAAGTACT TCGACACCAC CATCGACCGG AAGCGTTACA CGTCGACCAA GGAGGTCCTC
     5461 GACGCAACCC TTATCCACCA GTCCATCACC GGGCTTTACG AGACCCGTAT CGACCTTAGC
     5521 CAGCTTGGGG GGGATTAGCG GCCGCATAAT GATGTGTTAT CATTGATGCG AGGTCGCCTA
     5581 TACCTCCCCG CTTCGGCGGG GTTTTTTCCC GGGTTTACAC TTTATGCTTC CGGCTCGTAT
     5641 AATGGATGGC AGGTGCCCCA ACCCTACCCA CCTGCTAGAG TTTTAGAGCT AGAAATAGCA
     5701 AGTTAAAATA AGGCTAGTCC GTTATCAACT TGAAAAAGTG GCACCGAGTC GGTGCTTTga
     5761 tccggtggat gaccttttga atgaccttta atagattata ttactaatta attggggacc
     5821 ctagaggtcc ccttttttat tttaaaaatt ttttcacaaa acggtttaca agcataaagc
     5881 ttgctcaatc aatcaccGtc gacctcGTCT CCCCGCTTCG GCGGGGTTTT TTCCTAGGTT
     5941 GTGTCTCAAA ATCTCTGATG TTACATTGCA CAAGATAAAA ATATATCATC ATGAACAATA
     6001 AAACTGTCTG CTTACATAAA CAGTAATACA AGGGGTGTTA TGAGCCATAT TCAGCGTGAA
     6061 ACGAGCTGTA GCCGTCCGCG TCTGAACAGC AACATGGATG CGGATCTGTA TGGCTATAAA
     6121 TGGGCGCGTG ATAACGTGGG TCAGAGCGGC GCGACCATTT ATCGTCTGTA TGGCAAACCG
     6181 GATGCGCCGG AACTGTTTCT GAAACATGGC AAAGGCAGCG TGGCGAACGA TGTGACCGAT
     6241 GAAATGGTGC GTCTGAACTG GCTGACCGAA TTTATGCCGC TGCCGACCAT TAAACATTTT
     6301 ATTCGCACCC CGGATGATGC GTGGCTGCTG ACCACCGCGA TTCCGGGCAA AACCGCGTTT
     6361 CAGGTGCTGG AAGAATATCC GGATAGCGGC GAAAACATTG TGGATGCGCT GGCCGTGTTT
     6421 CTGCGTCGTC TGCATAGCAT TCCGGTGTGC AACTGCCCGT TTAACAGCGA TCGTGTGTTT
     6481 CGTCTGGCCC AGGCGCAGAG CCGTATGAAC AACGGCCTGG TGGATGCGAG CGATTTTGAT
     6541 GATGAACGTA ACGGCTGGCC GGTGGAACAG GTGTGGAAAG AAATGCATAA ACTGCTGCCG
     6601 TTTAGCCCGG ATAGCGTGGT GACCCACGGC GATTTTAGCC TGGATAACCT GATTTTCGAT
     6661 GAAGGCAAAC TGATTGGCTG CATTGATGTG GGCCGTGTGG GCATTGCGGA TCGTTATCAG
     6721 GATCTGGCCA TTCTGTGGAA CTGCCTGGGC GAATTTAGCC CGAGCCTGCA AAAACGTCTG
     6781 TTTCAGAAAT ATGGCATTGA TAATCCGGAT ATGAACAAAC TGCAATTTCA TCTGATGCTG
     6841 GATGAATTTT TCTAATAATT AATTGGACCG CGGTCCGAGA GACGCATGGC AGTATCTCAG
     6901 ACTGTGTATC GGCACGTCTC ACTACTTACC GGCGCGGCAG CGTTACCCGT GTCGGCGGCT
     6961 CCAACGGCTC GCCATCGTCC AGAAAACACG GCTCATCGGG CATCGGCAGG CGCTGCTGCC
     7021 CGCGCCGTTC CCATTCCTCC GTTTCGGTCA AGGCTGGCAG GTCTGGTTCC ATGCCCGGAA
     7081 TGCCGGGCTG GCTGGGCGGC TCCTCGCCGG GGCCGGTCGG TAGTTGCTGC TCGCCCGGAT
     7141 ACAGGGTCGG GATGCGGCGC AGGTCGCCAT GCCCCAACAG CGATTCGTCC TGGTCGTCGT
     7201 GATCAACCAC CACGGCGGCA CTGAACACCG ACAGGCGCAA CTGGTCGCGG GGCTGGCCCC
     7261 ACGCCACGCG G
//