LOCUS       sequence-248401-        6740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Basic Part O- ORF; Catalytically dead mutant of the Cas9 from 
            Streptococcus pyogenes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6740)
  AUTHORS   Valenzuela-Ortega M, French C
  TITLE     Joint universal modular plasmids (JUMP): a flexible vector platform 
            for synthetic biology.
  JOURNAL   Synth Biol (Oxf). 2021 Feb 2;6(1):ysab003. doi: 
            10.1093/synbio/ysab003. eCollection 2021.
  PUBMED    33623824
REFERENCE   2  (bases 1 to 6740)
  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: "Synth Biol 
            (Oxf)."; date: "2021-02-2"; volume: "6(1):ysab003. doi"; pages: " 
            10.1093/synbio/ysab003. eCollection 2021"
COMMENT     Sequence Label: pJUMP18-dCas9_O
FEATURES             Location/Qualifiers
     source          1..6740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    39..60
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             join(86..109,110..112,113..2599,2600..2602,2603..4192)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGDS"
     misc_feature    4210..4230
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      4264..4358
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        4431..4522
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4523..4591,4592..5383)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      5387..5426
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     oriT            5573..5681
                     /note="incP origin of transfer"
     rep_origin      5717..6305
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6206..6225
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6459..6476
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6593..6614
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      complement(6648..6734)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaattaagg agttttgcag gtgcacctgc ttttcgctga attcgcggcc gcttctagag
       61 ggtctgcgat gtttggtctc tagccgataa gaaatactca ataggcttag ctatcggcac
      121 aaatagcgtc ggatgggcgg tgatcactga tgaatataag gttccgtcta aaaagttcaa
      181 ggttctggga aatacagacc gccacagtat caaaaaaaat cttatagggg ctcttttatt
      241 tgacagtgga gagacagcgg aagcgactcg tcttaaacgg acagctcgta gaaggtatac
      301 acgtcggaag aatcgtattt gttatctaca ggagattttt tcaaatgaga tggcgaaagt
      361 agatgatagt ttctttcatc gacttgaaga gtcttttttg gtggaagaag ataagaagca
      421 tgaacgtcat cctatttttg gaaatatagt agatgaagtt gcttatcatg agaaatatcc
      481 aactatctat catctgcgaa aaaaattggt agattctact gataaagcgg atttgcgctt
      541 aatctatttg gccttagcgc atatgattaa gtttcgtggt cattttttga ttgagggaga
      601 tttaaatcct gataatagtg atgtggacaa actatttatc cagttggtac aaacctacaa
      661 tcaattattt gaagaaaacc ctattaacgc aagtggagta gatgctaaag cgattctttc
      721 tgcacgattg agtaaatcaa gacgattaga aaatctcatt gctcagctcc ccggtgagaa
      781 gaaaaatggc ttatttggga atctcattgc tttgtcattg ggtttgaccc ctaattttaa
      841 atcaaatttt gatttggcag aagatgctaa attacagctt tcaaaagata cttacgatga
      901 tgatttagat aatttattgg cgcaaattgg agatcaatat gctgatttgt ttttggcagc
      961 taagaattta tcagatgcta ttttactttc agatatccta agagtaaata ctgaaataac
     1021 taaggctccc ctatcagctt caatgattaa acgctacgat gaacatcatc aagacttgac
     1081 tcttttaaaa gctttagttc gacaacaact tccagaaaag tataaagaaa tcttttttga
     1141 tcaatcaaaa aacggatatg caggttatat tgatggggga gctagccaag aagaatttta
     1201 taaatttatc aaaccaattt tagaaaaaat ggatggtact gaggaattat tggtgaaact
     1261 aaatcgtgaa gatttgctgc gcaagcaacg gacctttgac aacggctcta ttccccatca
     1321 aattcacttg ggtgagctgc atgctatttt gagaaggcaa gaagattttt atccattttt
     1381 aaaagacaat cgtgagaaga ttgaaaaaat cttgactttt cgcattcctt attatgttgg
     1441 tccattggcg cgtggcaata gtcgttttgc atggatgact cggaagtctg aagaaacaat
     1501 taccccatgg aattttgaag aagttgtcga taaaggtgct tcagctcaat catttattga
     1561 acgcatgaca aactttgata aaaatcttcc aaatgaaaaa gtactaccaa aacatagttt
     1621 gctttatgag tattttacgg tttataacga attgacaaag gtcaaatatg ttactgaagg
     1681 aatgcgaaaa ccagcatttc tttcaggtga acagaagaaa gccattgttg atttactctt
     1741 caaaacaaat cgaaaagtaa ccgttaagca attaaaagaa gattatttca aaaaaataga
     1801 atgttttgat agtgttgaaa tttcaggagt tgaagataga tttaatgctt cattaggtac
     1861 ctaccatgat ttgctaaaaa ttattaaaga taaagatttt ttggataatg aagaaaatga
     1921 agatatctta gaggatattg ttttaacatt gaccttattt gaagataggg agatgattga
     1981 ggaaagactt aaaacatatg ctcacctctt tgatgataag gtgatgaaac agcttaaacg
     2041 tcgccgttat actggttggg gacgtttgtc tcgaaaattg attaatggta ttagggataa
     2101 gcaatctggc aaaacaatat tagatttttt gaaatcagat ggttttgcca atcgcaattt
     2161 tatgcagctg atccatgatg atagtttgac atttaaagaa gatattcaaa aagcacaagt
     2221 gtctggacaa ggcgatagtt tacatgaaca tattgcaaat ttagctggta gccctgctat
     2281 taaaaaaggt attttacaga ctgtaaaagt tgttgatgaa ttggtcaaag taatggggcg
     2341 gcataagcca gaaaatatcg ttattgaaat ggcacgtgaa aatcagacaa ctcaaaaggg
     2401 ccagaaaaat tcgcgagagc gtatgaaacg aatcgaagaa ggtatcaaag aattaggaag
     2461 tcagattctt aaagagcatc ctgttgaaaa tactcaattg caaaatgaaa agctctatct
     2521 ctattatctc caaaatggaa gagacatgta tgtggaccaa gaattagata ttaatcgttt
     2581 aagtgattat gatgtcgatg ccattgttcc acaaagtttc cttaaagacg attcaataga
     2641 caataaggtc ttaacgcgtt ctgataaaaa tcgtggtaaa tcggataacg ttccaagtga
     2701 agaagtagtc aaaaagatga aaaactattg gagacaactt ctaaacgcca agttaatcac
     2761 tcaacgtaag tttgataatt taacgaaagc tgaacgtgga ggtttgagtg aacttgataa
     2821 agctggtttt atcaaacgcc aattggttga aactcgccaa atcactaagc atgtggcaca
     2881 aattttggat agtcgcatga atactaaata cgatgaaaat gataaactta ttcgagaggt
     2941 taaagtgatt accttaaaat ctaaattagt ttctgacttc cgaaaagatt tccaattcta
     3001 taaagtacgt gagattaaca attaccatca tgcccatgat gcgtatctaa atgccgtcgt
     3061 tggaactgct ttgattaaga aatatccaaa acttgaatcg gagtttgtct atggtgatta
     3121 taaagtttat gatgttcgta aaatgattgc taagtctgag caagaaatag gcaaagcaac
     3181 cgcaaaatat ttcttttact ctaatatcat gaacttcttc aaaacagaaa ttacacttgc
     3241 aaatggagag attcgcaaac gccctctaat cgaaactaat ggggaaactg gagaaattgt
     3301 ctgggataaa gggcgagatt ttgccacagt gcgcaaagta ttgtccatgc cccaagtcaa
     3361 tattgtcaag aaaacagaag tacagacagg cggattctcc aaggagtcaa ttttaccaaa
     3421 aagaaattcg gacaagctta ttgctcgtaa aaaagactgg gatccaaaaa aatatggtgg
     3481 ttttgatagt ccaacggtag cttattcagt cctagtggtt gctaaggtgg aaaaagggaa
     3541 atcgaagaag ttaaaatccg ttaaagagtt actagggatc acaattatgg aaagaagttc
     3601 ctttgaaaaa aatccgattg actttttaga agctaaagga tataaggaag ttaaaaaaga
     3661 cttaatcatt aaactaccta aatatagtct ttttgagtta gaaaacggtc gtaaacggat
     3721 gctggctagt gccggagaat tacaaaaagg aaatgagctg gctctgccaa gcaaatatgt
     3781 gaatttttta tatttagcta gtcattatga aaagttgaag ggtagtccag aagataacga
     3841 acaaaaacaa ttgtttgtgg agcagcataa gcattattta gatgagatta ttgagcaaat
     3901 cagtgaattt tctaagcgtg ttattttagc agatgccaat ttagataaag ttcttagtgc
     3961 atataacaaa catagagaca aaccaatacg tgaacaagca gaaaatatta ttcatttatt
     4021 tacgttgacg aatcttggag ctcccgctgc ttttaaatat tttgatacaa caattgatcg
     4081 taaacgatat acgtctacaa aagaagtttt agatgccact cttatccatc aatccatcac
     4141 tggtctttat gaaacacgca ttgatttgag tcagctagga ggtgattcgt gagacctttc
     4201 atcgcgacct actagtagcg gccgctgcag ggagttgtct tcgaagactt cgctctagtc
     4261 ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa
     4321 cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aggggtcccc aataattacg
     4381 atttaaatta gtagcccgcc taatgagcgg gctttttttt aattccccta tttgtttatt
     4441 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4501 ataatattga aaaaggaaga gtatgagcat tcagcatttt cgtgtggcgc tgattccgtt
     4561 ttttgcggcg ttttgcctgc cggtgtttgc gcatccggaa accctggtga aagtgaaaga
     4621 tgcggaagat caactgggtg cgcgcgtggg ctatattgaa ctggatctga acagcggcaa
     4681 aattctggaa tcttttcgtc cggaagaacg ttttccgatg atgagcacct ttaaagtgct
     4741 gctgtgcggt gcggttctga gccgtgtgga tgcgggccag gaacaactgg gccgtcgtat
     4801 tcattatagc cagaacgatc tggtggaata tagcccggtg accgaaaaac atctgaccga
     4861 tggcatgacc gtgcgtgaac tgtgcagcgc ggcgattacc atgagcgata acaccgcggc
     4921 gaacctgctg ctgacgacca ttggcggtcc gaaagaactg accgcgtttc tgcataacat
     4981 gggcgatcat gtgacccgtc tggatcgttg ggaaccggaa ctgaacgaag cgattccgaa
     5041 cgatgaacgt gataccacca tgccggcagc aatggcgacc accctgcgta aactgctgac
     5101 gggtgagctg ctgaccctgg caagccgcca gcaactgatt gattggatgg aagcggataa
     5161 agtggcgggt ccgctgctgc gtagcgcgct gccggctggc tggtttattg cggataaaag
     5221 cggtgcgggc gaacgtggca gccgtggcat tattgcggcg ctgggcccgg atggtaaacc
     5281 gagccgtatt gtggtgattt ataccaccgg cagccaggcg acgatggatg aacgtaaccg
     5341 tcagattgcg gaaattggcg cgagcctgat taaacattgg taaaccgata caattaaagg
     5401 ctccttttgg agcctttttt tttggacgac ccttgtcctt ttccgctgca taaccctgct
     5461 tcggggtcat tatagcgatt ttttcggtat atccatcctt tttcgcacga tatacaggat
     5521 tttgccaaag ggttcgtgta gactttcctt ggtgtatcca acggcgtcag ccgggcagga
     5581 taggtgaagt aggcccaccc gcgagcgggt gttccttctt cactgtccct tattcgcacc
     5641 tggcggtgct caacgggaat cctgctctgc gaggctggcc gtaggccggc cccgtagaaa
     5701 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5761 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5821 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5881 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5941 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6001 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6061 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6121 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6181 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6241 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6301 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6361 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     6421 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     6481 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     6541 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     6601 gttagctcac tcattaggca ggcgcgccca gctgtctagg gcggcggatt tgtcctactc
     6661 aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttt cgactgagcc
     6721 tttcgtttta tttgatgcct
//
