pOP576

85943 Synthetic Bacterial Expression

Plasmid Information

Purpose
Temperature sensitive plasmid based on pDK46. Expresses dCas9 from proD. Serves as a control for pOP587 - pOP594
Depositor
Timothy Lu
Article
Purcell et al Nat Commun. 2018 Nov 22;9(1):4942. doi:
Insert
dCas9
Available Since
July 25, 2025
Availability
Available

Sequence Information

Length
7,321 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 3
Label Gene Product Position Strand
dCas9 — catalytically dead mutant of the Cas9 e… 206..4313 Fwd
DNA Sequence
ATGGATAAGAAATACTCTATTGGTTTGGCTATCGGTACAAACTCTGTTGGTTGGGCTGTTATTACTGATGAATACAAGGTTCCATCCAAGAAGTTCAAGGTTTTGGGTAACACTGATAGACACTCCATCAAAAAGAACTTGATTGGTGCCTTGTTGTTCGATTCTGGTGAAACTGCTGAAGCTACTAGATTGAAAAGAACCGCTAGAAGAAGATACACCAGAAGAAAGAACAGAATCTGCTACTTGCAAGAAATCTTCTCCAACGAAATGGCCAAGGTTGATGATTCATTCTTCCACAGATTGGAAGAATCCTTCTTGGTCGAAGAAGATAAGAAGCACGAAAGACATCCAATCTTCGGTAACATCGTTGATGAAGTTGCTTACCACGAAAAGTACCCAACTATCTACCATTTGAGAAAGAAGTTGGTTGACTCTACCGATAAGGCTGATTTGAGATTGATCTATTTGGCTTTGGCCCACATGATTAAGTTCAGAGGTCATTTCTTGATCGAAGGTGATTTGAACCCAGATAACTCCGATGTTGATAAGTTGTTCATCCAATTAGTCCAAACCTACAATCAATTATTCGAAGAAAACCCAATCAACGCCTCTGGTGTTGATGCTAAAGCTATTTTGTCTGCCAGATTGTCCAAGTCCAGAAGATTAGAAAATTTGATCGCCCAATTACCAGGTGAAAAGAAGAATGGTTTGTTCGGTAATTTGATTGCCTTGTCTTTGGGTTTGACTCCAAACTTCAAGTCCAATTTCGATTTGGCTGAAGATGCCAAGTTGCAATTATCTAAGGATACCTACGATGACGATTTGGATAACTTGTTGGCTCAAATCGGTGATCAATACGCTGATTTGTTTTTGGCTGCTAAGAACTTGTCCGATGCCATTTTGTTGTCCGATATTTTGAGAGTCAACACCGAAATTACTAAGGCTCCATTGTCTGCCTCTATGATCAAAAGATACGATGAACACCACCAAGACTTGACTTTGTTGAAGGCTTTGGTCAGACAACAATTACCTGAAAAGTACAAAGAAATTTTCTTCGATCAATCCAAGAACGGTTACGCCGGTTATATTGATGGTGGTGCTTCTCAAGAAGAATTTTACAAGTTCATCAAGCCAATCTTGGAAAAGATGGACGGTACTGAAGAATTATTGGTCAAGTTGAACAGAGAAGATTTGTTGAGAAAGCAAAGAACCTTCGACAACGGTTCTATTCCACATCAAATTCACTTGGGTGAATTGCACGCAATTTTGAGAAGACAAGAAGATTTTTATCCATTCTTGAAGGACAACAGAGAAAAGATCGAAAAGATTCTGACCTTCAGAATCCCTTACTACGTTGGTCCATTGGCTAGAGGTAATTCAAGATTTGCCTGGATGACTAGAAAGTCCGAAGAAACTATTACTCCTTGGAACTTCGAAGAAGTTGTAGATAAGGGTGCTTCTGCCCAATCCTTTATTGAAAGAATGACCAACTTCGACAAGAACTTGCCAAACGAAAAGGTTTTGCCAAAGCACTCTTTGTTGTACGAATACTTCACCGTCTACAACGAATTGACTAAGGTTAAGTACGTCACCGAAGGTATGAGAAAACCAGCTTTTTTATCCGGTGAACAAAAGAAGGCTATCGTCGATTTGTTGTTCAAGACCAACAGAAAGGTTACTGTCAAGCAATTAAAAGAAGATTACTTCAAGAAAATCGAATGCTTCGACTCCGTTGAAATTTCTGGTGTCGAAGATAGATTCAATGCCTCTTTAGGTACTTACCATGACTTGTTGAAAATCATCAAGGACAAGGATTTCTTGGACAACGAAGAAAACGAAGATATTTTGGAAGATATTGTCTTGACATTGACCTTGTTTGAAGATAGAGAAATGATTGAAGAAAGATTGAAAACCTACGCCCACTTGTTCGATGATAAGGTTATGAAGCAATTAAAGAGAAGAAGATACACTGGTTGGGGTAGATTGTCCAGAAAATTGATTAACGGTATCAGAGACAAGCAATCCGGTAAGACCATTTTGGACTTTTTGAAGTCTGATGGTTTCGCTAACAGAAACTTCATGCAATTAATCCACGACGATTCCTTGACTTTCAAAGAAGATATACAAAAGGCCCAAGTCTCTGGTCAAGGTGATTCTTTACATGAACATATCGCTAACTTGGCTGGTTCTCCAGCTATTAAGAAGGGTATTTTACAAACCGTTAAGGTCGTTGACGAATTGGTCAAAGTTATGGGTAGACATAAGCCAGAAAACATCGTTATCGAAATGGCTAGAGAAAATCAAACCACCCAAAAGGGTCAAAAGAACTCCAGAGAAAGAATGAAGAGAATCGAAGAAGGTATCAAAGAATTGGGTTCCCAAATTTTGAAAGAACACCCAGTTGAAAACACCCAATTACAAAACGAAAAGTTGTACTTGTACTACTTGCAAAACGGTAGAGATATGTACGTTGACCAAGAATTGGACATCAACAGATTGTCTGATTACGATGTTGACGCTATCGTTCCACAATCTTTTTTGAAGGATGACTCCATTGACAACAAGGTCTTGACTAGATCCGATAAGAATAGAGGTAAGTCCGATAACGTTCCATCTGAAGAAGTCGTTAAGAAAATGAAGAACTATTGGAGACAATTATTGAACGCCAAGTTGATCACCCAAAGAAAGTTTGACAATTTGACCAAGGCTGAAAGAGGTGGTTTGTCTGAATTGGATAAGGCAGGTTTCATCAAAAGACAATTAGTAGAAACCAGACAAATCACCAAGCACGTTGCTCAAATTTTGGATAGTAGAATGAACACTAAGTACGACGAAAACGACAAATTGATCAGAGAAGTTAAGGTCATTACCTTGAAGTCCAAGTTGGTTTCCGATTTCAGAAAGGACTTCCAATTCTACAAGGTCAGAGAAATCAACAACTACCATCATGCACATGATGCTTACTTGAATGCTGTTGTTGGTACTGCCTTGATTAAGAAGTATCCAAAGTTGGAATCCGAATTTGTCTACGGTGATTACAAGGTTTACGACGTTAGAAAGATGATCGCCAAGTCCGAACAAGAAATTGGTAAAGCTACTGCCAAATACTTCTTCTACTCCAATATTATGAATTTCTTTAAGACCGAAATCACTTTGGCCAACGGTGAAATTAGAAAAAGACCATTGATTGAAACTAATGGTGAAACAGGTGAAATCGTTTGGGATAAGGGTAGAGATTTTGCCACTGTTAGAAAGGTATTGTCCATGCCACAAGTAAACATCGTCAAAAAGACCGAAGTTCAAACTGGTGGTTTCTCCAAAGAATCCATTTTGCCTAAGAGAAACTCCGATAAGTTGATCGCTAGAAAAAAAGACTGGGACCCAAAAAAGTACGGTGGTTTTGATTCTCCAACTGTTGCTTACTCTGTTTTGGTTGTTGCTAAGGTCGAAAAGGGTAAGAGTAAGAAGTTGAAGTCCGTCAAAGAATTATTAGGTATCACTATCATGGAAAGATCCTCATTCGAAAAGAATCCTATCGACTTTTTGGAAGCCAAGGGTTACAAAGAAGTCAAGAAGGACTTGATCATTAAGTTGCCAAAGTACAGTTTGTTCGAATTGGAAAATGGTAGAAAGAGAATGTTGGCTTCTGCCGGTGAATTACAAAAGGGTAATGAATTGGCTTTGCCATCCAAGTACGTTAATTTCTTATACTTGGCCTCCCACTACGAAAAATTGAAAGGTTCTCCTGAAGATAACGAACAAAAGCAATTATTTGTCGAACAACACAAGCACTACTTGGACGAAATCATTGAACAAATTTCCGAATTTTCCAAAAGAGTCATTTTGGCTGACGCCAATTTGGACAAAGTTTTGTCAGCTTACAACAAGCACAGAGATAAGCCAATTAGAGAACAAGCTGAAAACATCATTCACTTGTTCACTTTGACTAACTTGGGTGCTCCAGCTGCTTTTAAGTATTTCGATACCACTATCGACAGAAAGAGATACACCTCTACCAAAGAAGTTTTGGACGCTACTTTGATCCACCAATCTATTACTGGTTTGTACGAAACTAGAATCGACTTGTCTCAATTAGGTGGTGATTAA
Protein Sequence
MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD
Rep101(Ts) rep101 encoding an A56V mutation temperature-sensitive version of the Re… 4487..5438 Rev
DNA Sequence
ATGTCTGAATTAGTTGTTTTCAAAGCAAATGAACTAGCGATTAGTCGCTATGACTTAACGGAGCATGAAACCAAGCTAATTTTATGCTGTGTGGCACTACTCAACCCCACGATTGAAAACCCTACAAGGAAAGAACGGACGGTATCGTTCACTTATAACCAATACGTTCAGATGATGAACATCAGTAGGGAAAATGCTTATGGTGTATTAGCTAAAGCAACCAGAGAGCTGATGACGAGAACTGTGGAAATCAGGAATCCTTTGGTTAAAGGCTTTGAGATTTTCCAGTGGACAAACTATGCCAAGTTCTCAAGCGAAAAATTAGAATTAGTTTTTAGTGAAGAGATATTGCCTTATCTTTTCCAGTTAAAAAAATTCATAAAATATAATCTGGAACATGTTAAGTCTTTTGAAAACAAATACTCTATGAGGATTTATGAGTGGTTATTAAAAGAACTAACACAAAAGAAAACTCACAAGGCAAATATAGAGATTAGCCTTGATGAATTTAAGTTCATGTTAATGCTTGAAAATAACTACCATGAGTTTAAAAGGCTTAACCAATGGGTTTTGAAACCAATAAGTAAAGATTTAAACACTTACAGCAATATGAAATTGGTGGTTGATAAGCGAGGCCGCCCGACTGATACGTTGATTTTCCAAGTTGAACTAGATAGACAAATGGATCTCGTAACCGAACTTGAGAACAACCAGATAAAAATGAATGGTGACAAAATACCAACAACCATTACATCAGATTCCTACCTACATAACGGACTAAGAAAAACACTACACGATGCTTTAACTGCAAAAATTCAGCTCACCAGTTTTGAGGCAAAATTTTTGAGTGACATGCAAAGTAAGTATGATCTCAATGGTTCGTTCTCATGGCTCACGCAAAAACAACGAACCACACTAGAGAACATACTGGCTAAATACGGAAGGATCTGA
Protein Sequence
MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKERTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFSSEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANIEISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQVELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKFLSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI
AmpR bla beta-lactamase 6329..7190 Rev
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
promoter 1
Label Gene Product Position Strand
AmpR promoter bla — 7190..7295 Rev
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
rep_origin 1
Label Gene Product Position Strand
pSC101 ori — — 5485..5708 Fwd
DNA Sequence
GGGCTAGTCAATGATAATTACTAGTCCTTTTCCTTTGAGTTGTGGGTATCTGTAAATTCTGCTAGACCTTTGCTGGAAAACTTGTAAATTCTGCTAGACCCTCTGTAAATTCCGCTAGACCTTTGTGTGTTTTTTTTGTTTATATTCAAGTGGTTATAATTTATAGAATAAAGAAAGAATAAAAAAAGATAAAAAGAATAGATCCCAGCCCTGTGTATAACTC