LOCUS       sequence-190101-        3407 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses the catalytically inactive version of ThermoCas9 
            (Thermo(d)Cas9) and its sgRNA module.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3407)
  AUTHORS   Mougiakos I, Mohanraju P, Bosma EF, Vrouwe V, Finger Bou M, 
            Naduthodi MIS, Gussak A, Brinkman RBL, van Kranenburg R, van der 
            Oost J
  TITLE     Characterizing a thermostable Cas9 for bacterial genome editing and 
            silencing.
  JOURNAL   Nat Commun. 2017 Nov 21;8(1):1647. doi: 10.1038/s41467-017-01591-4.
  PUBMED    29162801
REFERENCE   2  (bases 1 to 3407)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2017-11-21"; volume: "8(1):1647. doi"; pages: " 
            10.1038/s41467-017-01591-4"
COMMENT     Sequence Label: pThermoCas9i_ctrl
FEATURES             Location/Qualifiers
     source          1..3407
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(46..62)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(46..62)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(59..81)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    70..86
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(94..100,101..118,119..124))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    139..160
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(1047..2051)
                     /codon_start=1
                     /gene="repB"
                     /product="RepB replication protein"
                     /label=repB
                     /note="from Enterococcus faecalis plasmid pAM-alpha-1"
                     /db_xref="GI:22652809"
                     /protein_id="AAN03827.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
     primer_bind     complement(2572..2589)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2743..3331)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2823..2842)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 actataaaac tattgtgcat gggagggaag cttggcgtaa tcatggtcat agctgtttcc
       61 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
      121 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
      181 cgctttccag tcgggaaacc tgtcgtgcca gcccttcaaa cttcccaaag gcgagcccta
      241 gtgacattag aaaaccgact gtaaaaagta cagtcggcat tatctcatat tataaaagcc
      301 agtcattagg cctatctgac aattcctgaa tagagttcat aaacaatcct gcatgataac
      361 catcacaaac agaatgatgt acctgtaaag atagcggtaa atatattgaa ttacctttat
      421 taatgaattt tcctgctgta ataatgggta gaaggtaatt actattatta ttgatattta
      481 agttaaaccc agtaaatgaa gtccatggaa taatagaaag agaaaaagca ttttcaggta
      541 taggtgtttt gggaaacaat ttccccgaac cattatattt ctctacatca gaaaggtata
      601 aatcataaaa ctctttgaag tcattcttta caggagtcca aataccagag aatgttttag
      661 atacaccatc aaaaattgta taaagtggct ctaacttatc ccaataacct aactctccgt
      721 cgctattgta accagttcta aaagctgtat ttgagtttat cacccttgtc actaagaaaa
      781 taaatgcagg gtaaaattta tatccttctt gttttatgtt tcggtataaa acactaatat
      841 caatttctgt ggttatacta aaagtcgttt gttggttcaa ataatgatta aatatctctt
      901 ttctcttcca attgtctaaa tcaattttat taaagttcat ttgatatgcc tcctaaattt
      961 ttatctaaag tgaatttagg aggcttactt gtctgctttc ttcattagaa tcaatccttt
     1021 tttaaaagtc aatcccgttt gttgaactac tctttaataa aataattttt ccgttcccaa
     1081 ttccacattg caataataga aaatccatct tcatcggctt tttcgtcatc atctgtatga
     1141 atcaaatcgc cttcttctgt gtcatcaagg tttaattttt tatgtatttc ttttaacaaa
     1201 ccaccatagg agattaacct tttacggtgt aaaccttcct ccaaatcaga caaacgtttc
     1261 aaattctttt cttcatcatc ggtcataaaa tccgtatcct ttacaggata ttttgcagtt
     1321 tcgtcaattg ccgattgtat atccgattta tatttatttt tcggtcgaat catttgaact
     1381 tttacatttg gatcatagtc taatttcatt gcctttttcc aaaattgaat ccattgtttt
     1441 tgattcacgt agttttctgt attcttaaaa taagttggtt ccacacatac caatacatgc
     1501 atgtgctgat tataagaatt atctttatta tttattgtca cttccgttgc acgcataaaa
     1561 ccaacaagat ttttattaat ttttttatat tgcatcattc ggcgaaatcc ttgagccata
     1621 tctgacaaac tcttatttaa ttcttcgcca tcataaacat ttttaactgt taatgtgaga
     1681 aacaaccaac gaactgttgg cttttgttta ataacttcag caacaacctt ttgtgactga
     1741 atgccatgtt tcattgctct cctccagttg cacattggac aaagcctgga tttacaaaac
     1801 cacactcgat acaactttct ttcgcctgtt tcacgatttt gtttatactc taatatttca
     1861 gcacaatctt ttactctttc agccttttta aattcaagaa tatgcagaag ttcaaagtaa
     1921 tcaacattag cgattttctt ttctctccat ggtctcactt ttccactttt tgtcttgtcc
     1981 actaaaaccc ttgatttttc atctgaataa atgctactat taggacacat aatattaaaa
     2041 gaaaccccca tctatttagt tatttgtttg gtcacttata actttaacag atggggtttt
     2101 tctgtgcaac caattttaag ggttttcaat actttaaaac acatacatac caacacttca
     2161 acgcaccttt cagcaactaa aataaaaatg acgttatttc tatatgtatc aagaatagaa
     2221 agaactcgtt tttcgctacg ctcaaaacgc aaaaaaagca ctcattcgag tgctttttct
     2281 tatcgctcca aatcatgcga ttttttcctc tttgcttttc tttgctcacg aagttctcga
     2341 tcacgctgca aaacatcttg aagcgaaaaa gtattcttct tttcttccga tcgctcatgc
     2401 tgacgcacga aaagccctct aggcgcatag gaacaactcc taaatgcatg tgaggggttt
     2461 tctcgtccat gtgaacagtc gcatacgcaa tattttgttt cccatactgc attaatgaat
     2521 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2581 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2641 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2701 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2761 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2821 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2881 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2941 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3001 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3061 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3121 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3181 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3241 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3301 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3361 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatga
//
