pSET-dCas9-actII-4-NT-S1

110185 Synthetic Bacterial Expression

Plasmid Information

Purpose
Repression of gene expression in Streptomyces by CRISPRi
Depositor
Yinhua Lu
Article
Zhao et al Biotechnol J. 2018 Jun 3. doi: 10.1002/b
Mutation
D10A, H840A
Promoter
J23119 and ermE*p
Available Since
July 26, 2018
Availability
Available

Sequence Information

Length
10,400 bp
Topology
circular
Molecule Type
other DNA

Features

CDS 5
Label Gene Product Position Strand
dCas9 catalytically dead mutant of the Cas9 e… 665..4772 Fwd
DNA Sequence
ATGGACAAGAAGTACTCCATCGGCCTGGCGATCGGCACCAACTCCGTCGGCTGGGCCGTCATCACGGACGAGTACAAGGTCCCGTCGAAGAAGTTCAAGGTGCTGGGCAACACGGACCGCCACAGCATCAAGAAGAACCTCATCGGCGCCCTCCTGTTCGACAGCGGCGAGACGGCCGAGGCGACCCGCCTGAAGCGCACGGCGCGGCGGCGGTACACGCGCCGCAAGAACCGCATCTGCTACCTCCAGGAGATCTTCTCCAACGAGATGGCGAAGGTCGACGACTCGTTCTTCCACCGGCTGGAGGAGTCCTTCCTGGTCGAGGAGGACAAGAAGCACGAGCGGCACCCCATCTTCGGCAACATCGTGGACGAGGTGGCCTACCACGAGAAGTACCCGACCATCTACCACCTCCGGAAGAAGCTGGTGGACTCGACCGACAAGGCCGACCTGCGCCTGATCTACCTGGCCCTCGCCCACATGATCAAGTTCCGCGGCCACTTCCTCATCGAGGGCGACCTGAACCCGGACAACTCGGACGTCGACAAGCTGTTCATCCAGCTCGTGCAGACCTACAACCAGCTCTTCGAGGAGAACCCGATCAACGCCTCCGGCGTGGACGCGAAGGCCATCCTGTCGGCCCGCCTCTCGAAGTCCCGGCGGCTCGAGAACCTGATCGCCCAGCTGCCGGGCGAGAAGAAGAACGGCCTCTTCGGCAACCTGATCGCGCTCTCCCTGGGCCTGACGCCGAACTTCAAGTCCAACTTCGACCTCGCCGAGGACGCCAAGCTCCAGCTGTCCAAGGACACGTACGACGACGACCTGGACAACCTCCTGGCGCAGATCGGCGACCAGTACGCGGACCTGTTCCTCGCGGCGAAGAACCTCTCCGACGCGATCCTGCTCAGCGACATCCTCCGCGTCAACACCGAGATCACCAAGGCGCCCCTGTCCGCGAGCATGATCAAGCGCTACGACGAGCACCACCAGGACCTCACCCTGCTGAAGGCCCTGGTCCGCCAGCAGCTCCCGGAGAAGTACAAGGAGATCTTCTTCGACCAGTCCAAGAACGGCTACGCGGGCTACATCGACGGCGGCGCGTCCCAGGAGGAGTTCTACAAGTTCATCAAGCCCATCCTGGAGAAGATGGACGGCACCGAGGAGCTGCTGGTCAAGCTGAACCGGGAGGACCTCCTGCGCAAGCAGCGCACGTTCGACAACGGCTCCATCCCCCACCAGATCCACCTGGGCGAGCTGCACGCCATCCTCCGGCGCCAGGAGGACTTCTACCCGTTCCTCAAGGACAACCGCGAGAAGATCGAGAAGATCCTCACGTTCCGGATCCCGTACTACGTCGGCCCCCTCGCCCGCGGCAACTCCCGCTTCGCGTGGATGACGCGCAAGTCGGAGGAGACGATCACCCCGTGGAACTTCGAGGAGGTGGTGGACAAGGGCGCCTCCGCGCAGTCGTTCATCGAGCGGATGACCAACTTCGACAAGAACCTGCCGAACGAGAAGGTGCTGCCCAAGCACAGCCTCCTGTACGAGTACTTCACGGTCTACAACGAGCTCACCAAGGTGAAGTACGTGACGGAGGGCATGCGGAAGCCGGCGTTCCTGTCCGGCGAGCAGAAGAAGGCCATCGTGGACCTGCTGTTCAAGACGAACCGGAAGGTCACCGTCAAGCAGCTGAAGGAGGACTACTTCAAGAAGATCGAGTGCTTCGACTCGGTCGAGATCAGCGGCGTGGAGGACCGCTTCAACGCGTCGCTGGGCACGTACCACGACCTCCTGAAGATCATCAAGGACAAGGACTTCCTCGACAACGAGGAGAACGAGGACATCCTGGAGGACATCGTGCTGACCCTGACGCTCTTCGAGGACCGGGAGATGATCGAGGAGCGCCTCAAGACCTACGCGCACCTGTTCGACGACAAGGTCATGAAGCAGCTCAAGCGGCGGCGGTACACCGGCTGGGGCCGGCTCTCCCGCAAGCTCATCAACGGCATCCGGGACAAGCAGTCGGGCAAGACCATCCTCGACTTCCTGAAGTCCGACGGCTTCGCCAACCGCAACTTCATGCAGCTGATCCACGACGACAGCCTGACGTTCAAGGAGGACATCCAGAAGGCGCAGGTGTCGGGCCAGGGCGACTCCCTGCACGAGCACATCGCGAACCTGGCCGGCAGCCCGGCCATCAAGAAGGGCATCCTGCAGACGGTCAAGGTGGTCGACGAGCTGGTGAAGGTGATGGGCCGCCACAAGCCCGAGAACATCGTCATCGAGATGGCCCGGGAGAACCAGACCACGCAGAAGGGCCAGAAGAACAGCCGCGAGCGCATGAAGCGGATCGAGGAGGGCATCAAGGAGCTCGGCTCGCAGATCCTCAAGGAGCACCCCGTCGAGAACACCCAGCTCCAGAACGAGAAGCTCTACCTCTACTACCTGCAGAACGGCCGGGACATGTACGTGGACCAGGAGCTCGACATCAACCGGCTGTCGGACTACGACGTGGACGCCATCGTCCCGCAGTCGTTCCTGAAGGACGACAGCATCGACAACAAGGTCCTCACCCGCTCGGACAAGAACCGGGGCAAGTCCGACAACGTCCCGTCGGAGGAGGTCGTGAAGAAGATGAAGAACTACTGGCGGCAGCTCCTCAACGCCAAGCTCATCACGCAGCGCAAGTTCGACAACCTGACGAAGGCGGAGCGGGGCGGCCTCAGCGAGCTCGACAAGGCGGGCTTCATCAAGCGGCAGCTGGTCGAGACGCGGCAGATCACGAAGCACGTGGCCCAGATCCTGGACTCGCGCATGAACACCAAGTACGACGAGAACGACAAGCTGATCCGCGAGGTCAAGGTCATCACCCTCAAGTCCAAGCTGGTGAGCGACTTCCGCAAGGACTTCCAGTTCTACAAGGTGCGGGAGATCAACAACTACCACCACGCCCACGACGCGTACCTGAACGCGGTGGTGGGCACGGCGCTGATCAAGAAGTACCCGAAGCTCGAGAGCGAGTTCGTCTACGGCGACTACAAGGTGTACGACGTGCGGAAGATGATCGCGAAGAGCGAGCAGGAGATCGGCAAGGCCACGGCGAAGTACTTCTTCTACAGCAACATCATGAACTTCTTCAAGACCGAGATCACCCTGGCGAACGGCGAGATCCGCAAGCGCCCGCTCATCGAGACCAACGGCGAGACGGGCGAGATCGTGTGGGACAAGGGCCGCGACTTCGCGACCGTCCGGAAGGTGCTGTCCATGCCCCAGGTCAACATCGTCAAGAAGACGGAGGTCCAGACCGGCGGCTTCAGCAAGGAGTCCATCCTGCCCAAGCGGAACAGCGACAAGCTGATCGCCCGGAAGAAGGACTGGGACCCCAAGAAGTACGGCGGCTTCGACTCCCCCACCGTCGCCTACTCGGTCCTCGTGGTCGCCAAGGTGGAGAAGGGCAAGAGCAAGAAGCTGAAGTCCGTGAAGGAGCTGCTGGGCATCACCATCATGGAGCGGTCGTCGTTCGAGAAGAACCCCATCGACTTCCTGGAGGCCAAGGGCTACAAGGAGGTGAAGAAGGACCTCATCATCAAGCTCCCCAAGTACTCCCTGTTCGAGCTGGAGAACGGCCGGAAGCGCATGCTCGCCAGCGCGGGCGAGCTGCAGAAGGGCAACGAGCTGGCGCTGCCGAGCAAGTACGTCAACTTCCTGTACCTCGCCTCGCACTACGAGAAGCTGAAGGGCTCGCCGGAGGACAACGAGCAGAAGCAGCTGTTCGTGGAGCAGCACAAGCACTACCTCGACGAGATCATCGAGCAGATCTCCGAGTTCTCCAAGCGCGTCATCCTGGCCGACGCGAACCTGGACAAGGTCCTGTCCGCGTACAACAAGCACCGCGACAAGCCGATCCGGGAGCAGGCCGAGAACATCATCCACCTGTTCACGCTGACCAACCTCGGCGCCCCCGCGGCCTTCAAGTACTTCGACACCACGATCGACCGGAAGCGGTACACGAGCACGAAGGAGGTGCTCGACGCCACCCTGATCCACCAGAGCATCACGGGCCTGTACGAGACCCGCATCGACCTCTCCCAGCTGGGCGGCGACTGA
Protein Sequence
MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD
ApmR aac(3)IV aminoglycoside 3-N-acetyltransferase ty… 6386..7187 Fwd
DNA Sequence
ATGTCATCAGCGGTGGAGTGCAATGTCGTGCAATACGAATGGCGAAAAGCCGAGCTCATCGGTCAGCTTCTCAACCTTGGGGTTACCCCCGGCGGTGTGCTGCTGGTCCACAGCTCCTTCCGTAGCGTCCGGCCCCTCGAAGATGGGCCACTTGGACTGATCGAGGCCCTGCGTGCTGCGCTGGGTCCGGGAGGGACGCTCGTCATGCCCTCGTGGTCAGGTCTGGACGACGAGCCGTTCGATCCTGCCACGTCGCCCGTTACACCGGACCTTGGAGTTGTCTCTGACACATTCTGGCGCCTGCCAAATGTAAAGCGCAGCGCCCATCCATTTGCCTTTGCGGCAGCGGGGCCACAGGCAGAGCAGATCATCTCTGATCCATTGCCCCTGCCACCTCACTCGCCTGCAAGCCCGGTCGCCCGTGTCCATGAACTCGATGGGCAGGTACTTCTCCTCGGCGTGGGACACGATGCCAACACGACGCTGCATCTTGCCGAGTTGATGGCAAAGGTTCCCTATGGGGTGCCGAGACACTGCACCATTCTTCAGGATGGCAAGTTGGTACGCGTCGATTATCTCGAGAATGACCACTGCTGTGAGCGCTTTGCCTTGGCGGACAGGTGGCTCAAGGAGAAGAGCCTTCAGAAGGAAGGTCCAGTCGGTCATGCCTTTGCTCGGTTGATCCGCTCCCGCGACATTGTGGCGACAGCCCTGGGTCAACTGGGCCGAGATCCGTTGATCTTCCTGCATCCGCCAGAGGCGGGATGCGAAGAATGCGATGCCGCTCGCCAGTCGATTGGC
Protein Sequence
MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRPLEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNVKRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLHLAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGHAFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG
traJ traJ oriT-recognizing protein 7620..7992 Rev
DNA Sequence
ATGGCTGATGAAACCAAGCCAACCAGGAAGGGCAGCCCACCTATCAAGGTGTACTGCCTTCCAGACGAACGAAGAGCGATTGAGGAAAAGGCGGCGGCGGCCGGCATGAGCCTGTCGGCCTACCTGCTGGCCGTCGGCCAGGGCTACAAAATCACGGGCGTCGTGGACTATGAGCACGTCCGCGAGCTGGCCCGCATCAATGGCGACCTGGGCCGCCTGGGCGGCCTGCTGAAACTCTGGCTCACCGACGACCCGCGCACGGCGCGGTTCGGTGATGCCACGATCCTCGCCCTGCTGGCGAAGATCGAAGAGAAGCAGGACGAGCTTGGCAAGGTCATGATGGGCGTGGTCCGCCCGAGGGCAGAGCCATGA
Protein Sequence
MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAVGQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEEKQDELGKVMMGVVRPRAEP
Factor Xa site Factor Xa recognition and cleavage site 8371..8383 Rev
DNA Sequence
ATAGAGGGGCGT
Protein Sequence
IEGR
phage phi-C31 integrase integrase from phage phi-C31 8562..10380 Fwd
DNA Sequence
GTGGACACGTACGCGGGTGCTTACGACCGTCAGTCGCGCGAGCGCGAAAATTCGAGCGCAGCAAGCCCAGCGACACAGCGTAGCGCCAACGAAGACAAGGCGGCCGACCTTCAGCGCGAAGTCGAGCGCGACGGGGGCCGGTTCAGGTTCGTCGGGCATTTCAGCGAAGCGCCGGGCACGTCGGCGTTCGGGACGGCGGAGCGCCCGGAGTTCGAACGCATCCTGAACGAATGCCGCGCCGGGCGGCTCAACATGATCATTGTCTATGACGTGTCGCGCTTCTCGCGCCTGAAGGTCATGGACGCGATTCCGATTGTCTCGGAATTGCTCGCCCTGGGCGTGACGATTGTTTCCACTCAGGAAGGCGTCTTCCGGCAGGGAAACGTCATGGACCTGATTCACCTGATTATGCGGCTCGACGCGTCGCACAAAGAATCTTCGCTGAAGTCGGCGAAGATTCTCGACACGAAGAACCTTCAGCGCGAATTGGGCGGGTACGTCGGCGGGAAGGCGCCTTACGGCTTCGAGCTTGTTTCGGAGACGAAGGAGATCACGCGCAACGGCCGAATGGTCAATGTCGTCATCAACAAGCTTGCGCACTCGACCACTCCCCTTACCGGACCCTTCGAGTTCGAGCCCGACGTAATCCGGTGGTGGTGGCGTGAGATCAAGACGCACAAACACCTTCCCTTCAAGCCGGGCAGTCAAGCCGCCATTCACCCGGGCAGCATCACGGGGCTTTGTAAGCGCATGGACGCTGACGCCGTGCCGACCCGGGGCGAGACGATTGGGAAGAAGACCGCTTCAAGCGCCTGGGACCCGGCAACCGTTATGCGAATCCTTCGGGACCCGCGTATTGCGGGCTTCGCCGCTGAGGTGATCTACAAGAAGAAGCCGGACGGCACGCCGACCACGAAGATTGAGGGTTACCGCATTCAGCGCGACCCGATCACGCTCCGGCCGGTCGAGCTTGATTGCGGACCGATCATCGAGCCCGCTGAGTGGTATGAGCTTCAGGCGTGGTTGGACGGCAGGGGGCGCGGCAAGGGGCTTTCCCGGGGGCAAGCCATTCTGTCCGCCATGGACAAGCTGTACTGCGAGTGTGGCGCCGTCATGACTTCGAAGCGCGGGGAAGAATCGATCAAGGACTCTTACCGCTGCCGTCGCCGGAAGGTGGTCGACCCGTCCGCACCTGGGCAGCACGAAGGCACGTGCAACGTCAGCATGGCGGCACTCGACAAGTTCGTTGCGGAACGCATCTTCAACAAGATCAGGCACGCCGAAGGCGACGAAGAGACGTTGGCGCTTCTGTGGGAAGCCGCCCGACGCTTCGGCAAGCTCACTGAGGCGCCTGAGAAGAGCGGCGAACGGGCGAACCTTGTTGCGGAGCGCGCCGACGCCCTGAACGCCCTTGAAGAGCTGTACGAAGACCGCGCGGCAGGCGCGTACGACGGACCCGTTGGCAGGAAGCACTTCCGGAAGCAACAGGCAGCGCTGACGCTCCGGCAGCAAGGGGCGGAAGAGCGGCTTGCCGAACTTGAAGCCGCCGAAGCCCCGAAGCTTCCCCTTGACCAATGGTTCCCCGAAGACGCCGACGCTGACCCGACCGGCCCTAAGTCGTGGTGGGGGCGCGCGTCAGTAGACGACAAGCGCGTGTTCGTCGGGCTCTTCGTAGACAAGATCGTTGTCACGAAGTCGACTACGGGCAGGGGGCAGGGAACGCCCATCGAGAAGCGCGCTTCGATCACGTGGGCGAAGCCGCCGACCGACGACGACGAAGACGACGCCCAGGACGGCACGGAAGACGTAGCGGCGTAG
Protein Sequence
MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDGGRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIPIVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQRELGGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWREIKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRILRDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQAWLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPGQHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERANLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAEAPKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPIEKRASITWAKPPTDDDEDDAQDGTEDVAA
promoter 2
Label Gene Product Position Strand
J23119(SpeI) promoter 4953..4988 Fwd
DNA Sequence
TTGACAGCTAGCTCAGTCCTAGGTATAATACTAGT
lac promoter 5240..5271 Rev
DNA Sequence
TTTACACTTTATGCTTCCGGCTCGTATGTTG
protein_bind 3
Label Gene Product Position Strand
lac operator 5216..5233 Fwd
DNA Sequence
TTGTTATCCGCTCACAA
CAP binding site 5285..5307 Fwd
DNA Sequence
ATGAGTGAGCTAACTCACATTA
phage phi-C31 attP 8453..8553 Fwd
DNA Sequence
CCCAGGTCAGAAGCGGTTTTCGGGAGTAGTGCCCCAACTGGGGTAACCTTTGAGTTCTCTCAGTTGGGGGCGTAGGGTCGCCGACATGACACAAGGGGTT
rep_origin 1
Label Gene Product Position Strand
ori 5594..6183 Rev
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 1
Label Gene Product Position Strand
lambda t0 terminator 5100..5135 Rev
DNA Sequence
TTCAGAACGCTCGGTTGCCGCCGGGCGTTTTTTAT