pIA33 — PlasmidDB

Plasmid Information

Purpose

E. coli-C. difficile shuttle vector for CRISPR interference (CRISPRi) in C. difficile; sgRNA targets rfp; Pxyl::dCas9-opt Pgdh::sgRNA-rfp

Depositor

Craig Ellermeier, David S. Weiss

Article

Muh et al J Bacteriol. 2019 Feb 11. pii: JB.00711-

Mutation

Base-pairing region targets red fluorescent prote…

Promoter

Pgdh and Pxyl

Available Since

Feb. 12, 2019

Availability

Available

Sequence Information

Length

12,940 bp

Topology

circular

Molecule Type

other DNA

View on AddGene.org

Features

CDS 2

Label	Gene	Product	Position	Strand
dCas9	—	catalytically dead mutant of the Cas9 e…	1787..5894	Fwd
DNA Sequence ATGGATAAAAAATATAGTATAGGATTAGCAATAGGAACAAATAGTGTAGGATGGGCAGTAATAACTGATGAATATAAAGTACCAAGTAAAAAATTTAAAGTATTAGGAAATACAGATAGACATAGTATAAAAAAAAATTTAATAGGTGCATTACTTTTTGATAGTGGTGAAACAGCAGAAGCAACAAGACTTAAAAGAACAGCAAGAAGAAGATATACTAGAAGAAAAAATAGAATATGTTATTTACAAGAAATATTTAGTAATGAAATGGCAAAAGTAGATGATAGTTTTTTTCATAGATTAGAAGAAAGTTTTTTAGTAGAAGAAGATAAAAAACACGAAAGACATCCAATATTTGGAAATATAGTAGATGAAGTAGCATATCATGAAAAATATCCAACAATATATCATTTAAGAAAAAAACTTGTAGATAGTACAGATAAAGCAGATTTAAGACTTATATATCTTGCATTAGCACATATGATAAAATTTAGAGGACATTTTCTTATAGAAGGTGATTTAAATCCAGATAATAGTGATGTAGATAAACTTTTTATACAATTAGTACAAACATATAATCAATTATTTGAAGAAAATCCAATAAATGCAAGTGGTGTAGATGCAAAAGCAATATTAAGTGCAAGATTAAGTAAAAGTAGAAGATTAGAAAATCTTATAGCACAATTACCTGGAGAAAAAAAAAATGGATTATTTGGTAATCTTATAGCTTTAAGTTTAGGATTAACACCAAATTTTAAATCAAATTTTGATCTTGCAGAAGATGCAAAACTTCAATTAAGTAAAGATACATATGATGATGATTTAGATAATTTACTTGCACAAATAGGTGATCAATATGCAGATTTATTTCTTGCAGCAAAAAATCTTTCTGATGCAATACTTTTAAGTGATATACTTAGAGTAAATACAGAAATAACAAAAGCACCATTATCTGCAAGTATGATAAAAAGATATGATGAACATCATCAAGATTTAACATTACTTAAAGCATTAGTAAGACAACAATTACCAGAAAAATATAAAGAAATTTTTTTTGATCAAAGTAAAAATGGATATGCAGGATATATAGATGGTGGTGCAAGTCAAGAAGAATTTTATAAATTTATTAAACCAATATTAGAAAAAATGGATGGAACAGAAGAATTATTAGTAAAACTTAATAGAGAAGATTTACTTAGAAAACAAAGAACATTTGATAATGGAAGTATACCACATCAAATACATTTAGGTGAATTACATGCAATATTAAGAAGACAAGAAGATTTTTATCCATTTCTTAAAGATAATAGAGAAAAAATTGAAAAAATACTTACATTTAGAATACCATATTATGTAGGACCATTAGCAAGAGGAAATAGTAGATTTGCTTGGATGACAAGAAAAAGTGAAGAAACAATAACACCATGGAATTTTGAAGAAGTAGTAGATAAAGGAGCATCTGCACAAAGTTTTATAGAAAGAATGACAAATTTTGATAAAAATCTTCCAAATGAAAAAGTATTACCAAAACATAGTTTATTATATGAATATTTTACAGTATATAATGAATTAACAAAAGTAAAATATGTAACAGAAGGTATGAGAAAACCAGCATTTTTAAGTGGTGAACAAAAAAAAGCTATAGTAGATTTATTATTTAAAACAAATAGAAAAGTAACAGTAAAACAATTAAAAGAAGATTATTTTAAAAAAATTGAATGTTTTGATAGTGTAGAAATATCTGGTGTAGAAGATAGATTTAATGCAAGTTTAGGAACATATCATGATTTACTTAAAATAATAAAGGATAAAGATTTTTTAGATAATGAAGAAAATGAAGATATTTTAGAAGATATAGTATTAACATTAACTTTATTTGAAGATAGAGAAATGATAGAAGAAAGATTAAAAACTTATGCTCATTTATTTGATGATAAAGTAATGAAACAACTTAAAAGAAGAAGATATACAGGTTGGGGAAGATTAAGTAGAAAACTTATAAATGGAATAAGAGATAAACAAAGTGGAAAAACAATATTAGATTTTCTTAAAAGTGATGGATTTGCAAATAGAAATTTTATGCAATTAATACATGATGATAGTTTAACATTTAAAGAAGATATTCAAAAAGCACAAGTAAGTGGACAAGGTGATAGTTTACATGAACATATAGCAAATTTAGCAGGATCTCCAGCAATAAAAAAAGGTATATTACAAACAGTAAAAGTAGTTGATGAATTAGTAAAAGTAATGGGTAGACATAAACCAGAAAATATAGTTATAGAAATGGCAAGAGAAAATCAAACAACACAAAAAGGACAAAAAAATAGTAGAGAAAGAATGAAAAGAATAGAAGAAGGTATAAAAGAATTAGGAAGTCAAATACTTAAAGAACATCCAGTAGAAAATACACAACTTCAAAATGAAAAACTTTATTTATATTATTTACAAAATGGAAGAGATATGTATGTAGATCAAGAATTAGATATAAATAGATTAAGTGATTATGATGTAGATGCAATAGTACCACAAAGTTTTCTTAAAGATGATTCTATAGATAATAAAGTATTAACAAGATCTGATAAAAATAGAGGTAAAAGTGATAATGTACCAAGTGAAGAAGTTGTAAAAAAAATGAAAAATTATTGGAGACAATTACTTAATGCAAAACTTATAACACAAAGAAAATTTGATAATCTTACAAAAGCAGAAAGAGGTGGATTATCTGAATTAGATAAAGCTGGATTTATAAAAAGACAATTAGTAGAAACTAGACAAATAACAAAACATGTAGCACAAATATTAGATAGTAGAATGAATACAAAATATGATGAAAATGATAAGCTTATAAGAGAAGTAAAAGTTATAACATTAAAAAGTAAATTAGTAAGTGATTTTAGAAAAGATTTTCAATTTTATAAAGTAAGAGAAATAAATAATTATCATCATGCACATGATGCATATCTTAATGCAGTAGTAGGAACAGCATTAATAAAAAAATATCCTAAATTAGAATCTGAATTTGTATATGGAGATTATAAAGTATATGATGTAAGAAAAATGATAGCAAAAAGTGAACAAGAAATAGGAAAAGCAACAGCAAAGTACTTTTTTTATTCTAATATAATGAATTTTTTTAAGACAGAAATAACTTTAGCAAATGGTGAAATAAGAAAAAGACCTTTAATAGAAACTAATGGTGAAACAGGTGAAATAGTATGGGATAAAGGTAGAGATTTTGCAACAGTAAGAAAAGTATTAAGTATGCCACAAGTAAATATAGTAAAAAAAACAGAAGTACAAACTGGAGGATTTAGTAAAGAAAGTATACTTCCAAAAAGAAATTCAGATAAACTTATTGCAAGAAAAAAAGATTGGGATCCAAAAAAATATGGTGGATTTGATAGTCCAACAGTAGCATATAGTGTATTAGTAGTAGCAAAAGTAGAAAAAGGTAAATCTAAAAAACTTAAAAGTGTAAAAGAATTACTTGGAATAACAATAATGGAAAGAAGTAGTTTTGAAAAAAATCCTATTGATTTTTTAGAAGCAAAAGGATATAAAGAAGTTAAAAAAGATTTAATAATAAAACTTCCAAAATATTCTTTATTTGAATTAGAAAATGGAAGAAAAAGAATGTTAGCAAGTGCTGGTGAATTACAAAAAGGTAATGAATTAGCATTACCATCTAAATATGTAAATTTTCTTTATTTAGCAAGTCATTATGAAAAACTTAAAGGATCTCCTGAAGATAATGAACAAAAACAATTATTTGTAGAACAACATAAACATTATTTAGATGAAATAATTGAACAAATAAGTGAATTTTCTAAAAGAGTAATATTAGCAGACGCAAATTTAGATAAAGTATTATCTGCTTATAATAAACATAGAGATAAACCAATAAGAGAACAAGCAGAAAATATAATACATCTTTTTACATTAACAAATTTAGGTGCACCAGCAGCATTTAAATATTTTGATACAACAATAGATAGAAAAAGATATACAAGTACAAAAGAAGTATTAGACGCAACATTAATACATCAAAGTATAACAGGATTATATGAAACAAGAATAGATTTATCTCAATTAGGTGGTGATTAA Protein Sequence MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD
traJ	traJ	oriT-recognizing protein	7257..7629	Rev
DNA Sequence ATGGCTGATGAAACCAAGCCAACCAGGAAGGGCAGCCCACCTATCAAGGTGTACTGCCTTCCAGACGAACGAAGAGCGATTGAGGAAAAGGCGGCGGCGGCCGGCATGAGCCTGTCGGCCTACCTGCTGGCCGTCGGCCAGGGCTACAAAATCACGGGCGTCGTGGACTATGAGCACGTCCGCGAGCTGGCCCGCATCAATGGCGACCTGGGCCGCCTGGGCGGCCTGCTGAAACTCTGGCTCACCGACGACCCGCGCACGGCGCGGTTCGGTGATGCCACGATCCTCGCCCTGCTGGCGAAGATCGAAGAGAAGCAGGACGAGCTTGGCAAGGTCATGATGGGCGTGGTCCGCCCGAGGGCAGAGCCATGA Protein Sequence MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAVGQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEEKQDELGKVMMGVVRPRAEP

rep_origin 1

Label	Gene	Product	Position	Strand
ori	—	—	9137..9726	Rev
DNA Sequence TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCAAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA