pCRISPRi_Mxi1_yl

91248 Synthetic Yeast Expression

Plasmid Information

Purpose
CRISPR-dCas9-Mxi1 vector for Yarrowia lipolytica, expressing dCas9-Mxi1 and AvrII site for sgRNA insertion
Depositor
Ian Wheeldon
Article
Schwartz et al Biotechnol Bioeng. 2017 Aug 19. doi: 10.
Promoter
SCR1'-tRNA and UAS1B8-TEF(136)
Available Since
Aug. 25, 2017
Availability
Available

Sequence Information

Length
11,977 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… 1739..5843 Fwd
DNA Sequence
ATGGATAAGAAATACTCCATTGGCCTGGCCATCGGAACCAACTCCGTGGGTTGGGCCGTGATCACCGATGAGTACAAGGTGCCCTCTAAGAAATTCAAGGTCCTGGGCAACACCGACCGACACTCCATCAAGAAGAACCTGATCGGCGCTCTGCTCTTCGACTCTGGCGAGACCGCTGAGGCCACCCGACTGAAGCGAACCGCTCGAAGACGATACACCCGAAGAAAGAACCGAATCTGTTACCTGCAGGAGATCTTCTCTAACGAGATGGCCAAGGTGGACGACTCTTTCTTCCACCGACTGGAGGAGTCTTTCCTGGTGGAGGAGGACAAGAAGCACGAGCGACACCCCATCTTCGGCAACATCGTGGACGAGGTGGCCTACCACGAGAAGTACCCCACCATCTACCACCTGCGAAAGAAGCTGGTGGACTCTACCGACAAGGCCGACCTGCGACTGATCTACCTGGCCCTGGCCCACATGATCAAGTTCCGAGGCCACTTCCTGATCGAGGGCGACCTGAACCCCGACAACTCTGACGTGGACAAGCTGTTCATCCAGCTGGTGCAGACCTACAACCAGCTCTTCGAAGAGAACCCCATTAACGCTTCTGGCGTGGATGCTAAGGCCATCCTGTCTGCCCGACTGTCTAAGTCTCGACGACTCGAGAACCTGATTGCTCAGCTCCCCGGAGAGAAGAAGAACGGTCTGTTCGGAAACCTGATTGCTCTGTCCCTGGGTCTCACCCCTAACTTCAAGTCCAACTTCGATCTGGCTGAGGACGCTAAGCTGCAGCTGTCTAAGGACACCTACGACGATGACCTGGATAACCTGCTCGCCCAGATTGGCGACCAGTACGCCGACCTGTTCCTGGCCGCCAAGAACCTGTCTGACGCCATCCTGCTGTCTGACATCCTGCGAGTGAACACCGAGATCACCAAGGCCCCCCTGTCTGCCTCCATGATTAAGCGATACGATGAGCACCACCAGGATCTGACCCTCCTCAAGGCTCTGGTCCGACAGCAGCTGCCCGAGAAGTACAAGGAGATTTTCTTCGACCAGTCTAAGAACGGCTACGCCGGCTACATCGACGGCGGCGCCTCTCAGGAGGAGTTCTACAAGTTCATTAAGCCCATCCTGGAGAAGATGGACGGAACCGAGGAACTGCTCGTGAAGCTGAACCGAGAGGACCTCCTGCGAAAGCAGCGAACCTTCGACAACGGCTCTATCCCCCACCAGATCCACCTGGGCGAGCTGCACGCCATCCTGCGACGACAGGAGGACTTCTACCCCTTCCTGAAGGACAACCGAGAGAAGATCGAGAAGATCCTGACCTTCCGAATCCCCTACTACGTGGGACCCCTGGCCCGAGGAAACTCTCGATTCGCTTGGATGACCCGAAAGTCTGAGGAGACCATTACCCCCTGGAACTTCGAGGAGGTGGTGGATAAGGGCGCCTCTGCTCAGTCTTTCATCGAGCGAATGACCAACTTCGACAAGAACCTCCCCAACGAGAAGGTCCTGCCCAAGCACTCTCTGCTCTACGAGTACTTCACCGTCTACAACGAGCTCACCAAGGTCAAGTACGTGACCGAGGGAATGCGAAAGCCCGCTTTCCTGTCTGGAGAGCAGAAGAAGGCTATTGTGGATCTGCTCTTCAAGACTAACCGAAAGGTCACCGTCAAGCAGCTGAAGGAGGATTACTTCAAGAAGATTGAGTGTTTCGATTCTGTCGAGATCTCCGGCGTCGAGGACCGATTCAACGCCTCTCTGGGTACCTACCACGACCTGCTGAAGATTATCAAGGACAAGGATTTCCTGGATAACGAGGAGAACGAGGATATTCTCGAGGACATTGTCCTGACCCTCACCCTGTTCGAGGATCGAGAGATGATTGAGGAGCGACTCAAGACCTACGCTCACCTGTTCGACGACAAGGTGATGAAGCAGCTGAAGCGACGACGATACACCGGCTGGGGCCGACTGTCTCGAAAGCTGATCAACGGCATCCGAGACAAGCAGTCTGGCAAGACCATCCTGGACTTCCTGAAGTCTGACGGCTTCGCCAACCGAAACTTCATGCAGCTGATCCACGACGACTCTCTGACCTTCAAGGAGGACATCCAGAAGGCCCAGGTGTCTGGCCAGGGCGACTCTCTGCACGAGCACATCGCCAACCTGGCCGGCTCTCCCGCCATTAAGAAAGGTATCCTGCAGACCGTCAAGGTGGTCGATGAGCTCGTCAAGGTGATGGGCCGACACAAGCCCGAGAACATTGTCATTGAGATGGCTCGAGAGAACCAGACTACTCAGAAGGGTCAGAAAAACTCCCGAGAGCGAATGAAGCGAATTGAGGAAGGTATTAAGGAGCTGGGATCCCAGATTCTCAAGGAGCATCCCGTGGAGAACACTCAGCTCCAGAACGAGAAGCTGTACCTGTACTATCTGCAGAACGGTCGAGACATGTACGTCGACCAGGAGCTGGATATCAACCGACTCTCCGACTACGATGTGGACGCCATTGTGCCCCAGTCCTTCCTGAAGGACGATTCTATCGATAACAAGGTGCTGACCCGATCCGACAAGAACCGAGGCAAGTCTGACAACGTGCCCTCCGAGGAGGTGGTCAAGAAGATGAAGAACTACTGGCGACAGCTGCTGAACGCCAAGCTGATTACCCAGCGAAAGTTCGACAACCTGACCAAGGCCGAGCGAGGCGGCCTGTCTGAGCTGGACAAGGCCGGCTTCATCAAGCGACAGCTGGTGGAGACCCGACAGATCACCAAGCACGTGGCCCAGATCCTGGACTCTCGAATGAACACCAAGTACGACGAGAACGACAAGCTGATCCGAGAGGTGAAGGTGATCACCCTGAAGTCTAAGCTGGTGTCTGACTTCCGAAAGGACTTCCAGTTCTACAAGGTGCGAGAGATTAACAACTACCACCACGCCCACGATGCCTACCTGAACGCTGTCGTGGGCACCGCCCTCATCAAGAAGTATCCCAAGCTGGAGTCCGAGTTCGTCTACGGCGACTACAAGGTCTACGATGTGCGAAAAATGATTGCCAAGTCCGAGCAGGAGATTGGCAAGGCTACCGCCAAGTACTTCTTCTACTCCAACATTATGAACTTCTTCAAGACCGAGATTACCCTGGCTAACGGCGAGATTCGAAAGCGACCCCTCATTGAGACCAACGGAGAGACCGGTGAGATCGTGTGGGACAAGGGACGAGACTTCGCCACCGTGCGAAAGGTGCTGTCTATGCCCCAGGTGAACATCGTGAAGAAGACCGAGGTGCAGACCGGAGGTTTCTCTAAGGAGTCCATCCTGCCCAAGCGAAACTCTGACAAGCTGATCGCCCGAAAGAAGGACTGGGACCCCAAGAAGTACGGAGGTTTCGACTCTCCCACCGTGGCTTACTCTGTGCTGGTGGTGGCCAAGGTGGAGAAGGGCAAGTCTAAGAAGCTGAAGTCTGTGAAGGAGCTGCTGGGCATTACCATCATGGAGCGATCTTCTTTCGAGAAGAACCCCATTGACTTCCTGGAGGCCAAGGGATACAAGGAGGTGAAGAAAGATCTGATTATCAAGCTCCCCAAGTACTCTCTGTTCGAGCTGGAGAACGGACGAAAGCGAATGCTGGCCTCTGCCGGCGAGCTGCAGAAGGGAAACGAGCTGGCCCTGCCCTCCAAGTACGTCAACTTCCTGTACCTCGCCTCCCATTACGAGAAGCTGAAGGGCTCTCCCGAGGATAACGAGCAGAAGCAGCTCTTCGTGGAGCAGCATAAGCACTACCTGGACGAGATCATCGAGCAGATCTCTGAGTTCTCTAAGCGAGTGATCCTGGCTGACGCCAACCTGGATAAGGTGCTGTCTGCTTACAACAAGCACCGAGACAAGCCCATTCGAGAGCAGGCTGAGAACATCATTCACCTGTTCACCCTGACCAACCTGGGAGCCCCCGCTGCCTTCAAGTACTTCGACACCACCATCGACCGAAAGCGATACACCTCTACCAAGGAGGTGCTGGACGCCACCCTGATCCACCAGTCTATCACCGGCCTGTACGAGACCCGAATCGACCTGTCTCAGCTGGGCGGCGAC
Protein Sequence
MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD
SV40 NLS — nuclear localization signal of SV40 (si… 5855..5876 Fwd
DNA Sequence
CCCAAGAAGAAGCGAAAGGTG
Protein Sequence
PKKKRKV
AmpR bla beta-lactamase 7484..8345 Rev
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
promoter 2
Label Gene Product Position Strand
lac promoter — — 6371..6402 Rev
DNA Sequence
TTTACACTTTATGCTTCCGGCTCGTATGTTG
AmpR promoter bla — 8345..8450 Rev
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
protein_bind 2
Label Gene Product Position Strand
lac operator — — 6347..6364 Fwd
DNA Sequence
TTGTTATCCGCTCACAA
CAP binding site — — 6416..6438 Fwd
DNA Sequence
ATGAGTGAGCTAACTCACATTA
rep_origin 1
Label Gene Product Position Strand
ori — — 6725..7314 Rev
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 1
Label Gene Product Position Strand
CYC1 terminator S. cerevisiae CYC1 — 6057..6305 Fwd
DNA Sequence
TCATGTAATTAGTTATGTCACGCTTACATTCACGCCCTCCCTCCACATCCGCTCTAACCGAAAAGGAAGGAGTTAGACAACCTGAAGTCTAGGTCCCTATTTATTTTTTTATAGTTATGTTAGTATTAAGAACGTTATTTATATTTCAAATTTTTCTTTTTTTTCTGTACAGACGCGTGTACGCATGTAACATTATACTGAAAACCTTGCTTGAGAAGGTTTTGGGACGCTCGAAGGCTTTAATTTGC