pCRISPRi_Mxi1_yl_NHEJ

91249 Synthetic Yeast Expression

Plasmid Information

Purpose
CRISPRi vector for Yarrowia lipolytica, repressing KU70 and KU80 for enhanced HR
Depositor
Ian Wheeldon
Article
Schwartz et al Biotechnol Bioeng. 2017 Aug 19. doi: 10.
Promoter
SCR1'-tRNA and UAS1B8-TEF(136)
Available Since
Aug. 24, 2017
Availability
Available

Sequence Information

Length
13,333 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… 3077..7181 Fwd
DNA Sequence
ATGGATAAGAAATACTCCATTGGCCTGGCCATCGGAACCAACTCCGTGGGTTGGGCCGTGATCACCGATGAGTACAAGGTGCCCTCTAAGAAATTCAAGGTCCTGGGCAACACCGACCGACACTCCATCAAGAAGAACCTGATCGGCGCTCTGCTCTTCGACTCTGGCGAGACCGCTGAGGCCACCCGACTGAAGCGAACCGCTCGAAGACGATACACCCGAAGAAAGAACCGAATCTGTTACCTGCAGGAGATCTTCTCTAACGAGATGGCCAAGGTGGACGACTCTTTCTTCCACCGACTGGAGGAGTCTTTCCTGGTGGAGGAGGACAAGAAGCACGAGCGACACCCCATCTTCGGCAACATCGTGGACGAGGTGGCCTACCACGAGAAGTACCCCACCATCTACCACCTGCGAAAGAAGCTGGTGGACTCTACCGACAAGGCCGACCTGCGACTGATCTACCTGGCCCTGGCCCACATGATCAAGTTCCGAGGCCACTTCCTGATCGAGGGCGACCTGAACCCCGACAACTCTGACGTGGACAAGCTGTTCATCCAGCTGGTGCAGACCTACAACCAGCTCTTCGAAGAGAACCCCATTAACGCTTCTGGCGTGGATGCTAAGGCCATCCTGTCTGCCCGACTGTCTAAGTCTCGACGACTCGAGAACCTGATTGCTCAGCTCCCCGGAGAGAAGAAGAACGGTCTGTTCGGAAACCTGATTGCTCTGTCCCTGGGTCTCACCCCTAACTTCAAGTCCAACTTCGATCTGGCTGAGGACGCTAAGCTGCAGCTGTCTAAGGACACCTACGACGATGACCTGGATAACCTGCTCGCCCAGATTGGCGACCAGTACGCCGACCTGTTCCTGGCCGCCAAGAACCTGTCTGACGCCATCCTGCTGTCTGACATCCTGCGAGTGAACACCGAGATCACCAAGGCCCCCCTGTCTGCCTCCATGATTAAGCGATACGATGAGCACCACCAGGATCTGACCCTCCTCAAGGCTCTGGTCCGACAGCAGCTGCCCGAGAAGTACAAGGAGATTTTCTTCGACCAGTCTAAGAACGGCTACGCCGGCTACATCGACGGCGGCGCCTCTCAGGAGGAGTTCTACAAGTTCATTAAGCCCATCCTGGAGAAGATGGACGGAACCGAGGAACTGCTCGTGAAGCTGAACCGAGAGGACCTCCTGCGAAAGCAGCGAACCTTCGACAACGGCTCTATCCCCCACCAGATCCACCTGGGCGAGCTGCACGCCATCCTGCGACGACAGGAGGACTTCTACCCCTTCCTGAAGGACAACCGAGAGAAGATCGAGAAGATCCTGACCTTCCGAATCCCCTACTACGTGGGACCCCTGGCCCGAGGAAACTCTCGATTCGCTTGGATGACCCGAAAGTCTGAGGAGACCATTACCCCCTGGAACTTCGAGGAGGTGGTGGATAAGGGCGCCTCTGCTCAGTCTTTCATCGAGCGAATGACCAACTTCGACAAGAACCTCCCCAACGAGAAGGTCCTGCCCAAGCACTCTCTGCTCTACGAGTACTTCACCGTCTACAACGAGCTCACCAAGGTCAAGTACGTGACCGAGGGAATGCGAAAGCCCGCTTTCCTGTCTGGAGAGCAGAAGAAGGCTATTGTGGATCTGCTCTTCAAGACTAACCGAAAGGTCACCGTCAAGCAGCTGAAGGAGGATTACTTCAAGAAGATTGAGTGTTTCGATTCTGTCGAGATCTCCGGCGTCGAGGACCGATTCAACGCCTCTCTGGGTACCTACCACGACCTGCTGAAGATTATCAAGGACAAGGATTTCCTGGATAACGAGGAGAACGAGGATATTCTCGAGGACATTGTCCTGACCCTCACCCTGTTCGAGGATCGAGAGATGATTGAGGAGCGACTCAAGACCTACGCTCACCTGTTCGACGACAAGGTGATGAAGCAGCTGAAGCGACGACGATACACCGGCTGGGGCCGACTGTCTCGAAAGCTGATCAACGGCATCCGAGACAAGCAGTCTGGCAAGACCATCCTGGACTTCCTGAAGTCTGACGGCTTCGCCAACCGAAACTTCATGCAGCTGATCCACGACGACTCTCTGACCTTCAAGGAGGACATCCAGAAGGCCCAGGTGTCTGGCCAGGGCGACTCTCTGCACGAGCACATCGCCAACCTGGCCGGCTCTCCCGCCATTAAGAAAGGTATCCTGCAGACCGTCAAGGTGGTCGATGAGCTCGTCAAGGTGATGGGCCGACACAAGCCCGAGAACATTGTCATTGAGATGGCTCGAGAGAACCAGACTACTCAGAAGGGTCAGAAAAACTCCCGAGAGCGAATGAAGCGAATTGAGGAAGGTATTAAGGAGCTGGGATCCCAGATTCTCAAGGAGCATCCCGTGGAGAACACTCAGCTCCAGAACGAGAAGCTGTACCTGTACTATCTGCAGAACGGTCGAGACATGTACGTCGACCAGGAGCTGGATATCAACCGACTCTCCGACTACGATGTGGACGCCATTGTGCCCCAGTCCTTCCTGAAGGACGATTCTATCGATAACAAGGTGCTGACCCGATCCGACAAGAACCGAGGCAAGTCTGACAACGTGCCCTCCGAGGAGGTGGTCAAGAAGATGAAGAACTACTGGCGACAGCTGCTGAACGCCAAGCTGATTACCCAGCGAAAGTTCGACAACCTGACCAAGGCCGAGCGAGGCGGCCTGTCTGAGCTGGACAAGGCCGGCTTCATCAAGCGACAGCTGGTGGAGACCCGACAGATCACCAAGCACGTGGCCCAGATCCTGGACTCTCGAATGAACACCAAGTACGACGAGAACGACAAGCTGATCCGAGAGGTGAAGGTGATCACCCTGAAGTCTAAGCTGGTGTCTGACTTCCGAAAGGACTTCCAGTTCTACAAGGTGCGAGAGATTAACAACTACCACCACGCCCACGATGCCTACCTGAACGCTGTCGTGGGCACCGCCCTCATCAAGAAGTATCCCAAGCTGGAGTCCGAGTTCGTCTACGGCGACTACAAGGTCTACGATGTGCGAAAAATGATTGCCAAGTCCGAGCAGGAGATTGGCAAGGCTACCGCCAAGTACTTCTTCTACTCCAACATTATGAACTTCTTCAAGACCGAGATTACCCTGGCTAACGGCGAGATTCGAAAGCGACCCCTCATTGAGACCAACGGAGAGACCGGTGAGATCGTGTGGGACAAGGGACGAGACTTCGCCACCGTGCGAAAGGTGCTGTCTATGCCCCAGGTGAACATCGTGAAGAAGACCGAGGTGCAGACCGGAGGTTTCTCTAAGGAGTCCATCCTGCCCAAGCGAAACTCTGACAAGCTGATCGCCCGAAAGAAGGACTGGGACCCCAAGAAGTACGGAGGTTTCGACTCTCCCACCGTGGCTTACTCTGTGCTGGTGGTGGCCAAGGTGGAGAAGGGCAAGTCTAAGAAGCTGAAGTCTGTGAAGGAGCTGCTGGGCATTACCATCATGGAGCGATCTTCTTTCGAGAAGAACCCCATTGACTTCCTGGAGGCCAAGGGATACAAGGAGGTGAAGAAAGATCTGATTATCAAGCTCCCCAAGTACTCTCTGTTCGAGCTGGAGAACGGACGAAAGCGAATGCTGGCCTCTGCCGGCGAGCTGCAGAAGGGAAACGAGCTGGCCCTGCCCTCCAAGTACGTCAACTTCCTGTACCTCGCCTCCCATTACGAGAAGCTGAAGGGCTCTCCCGAGGATAACGAGCAGAAGCAGCTCTTCGTGGAGCAGCATAAGCACTACCTGGACGAGATCATCGAGCAGATCTCTGAGTTCTCTAAGCGAGTGATCCTGGCTGACGCCAACCTGGATAAGGTGCTGTCTGCTTACAACAAGCACCGAGACAAGCCCATTCGAGAGCAGGCTGAGAACATCATTCACCTGTTCACCCTGACCAACCTGGGAGCCCCCGCTGCCTTCAAGTACTTCGACACCACCATCGACCGAAAGCGATACACCTCTACCAAGGAGGTGCTGGACGCCACCCTGATCCACCAGTCTATCACCGGCCTGTACGAGACCCGAATCGACCTGTCTCAGCTGGGCGGCGAC
Protein Sequence
MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD
SV40 NLS — nuclear localization signal of SV40 (si… 7193..7214 Fwd
DNA Sequence
CCCAAGAAGAAGCGAAAGGTG
Protein Sequence
PKKKRKV
AmpR bla beta-lactamase 8822..9683 Rev
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
promoter 2
Label Gene Product Position Strand
lac promoter — — 7709..7740 Rev
DNA Sequence
TTTACACTTTATGCTTCCGGCTCGTATGTTG
AmpR promoter bla — 9683..9788 Rev
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
protein_bind 2
Label Gene Product Position Strand
lac operator — — 7685..7702 Fwd
DNA Sequence
TTGTTATCCGCTCACAA
CAP binding site — — 7754..7776 Fwd
DNA Sequence
ATGAGTGAGCTAACTCACATTA
rep_origin 1
Label Gene Product Position Strand
ori — — 8063..8652 Rev
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 1
Label Gene Product Position Strand
CYC1 terminator S. cerevisiae CYC1 — 7395..7643 Fwd
DNA Sequence
TCATGTAATTAGTTATGTCACGCTTACATTCACGCCCTCCCTCCACATCCGCTCTAACCGAAAAGGAAGGAGTTAGACAACCTGAAGTCTAGGTCCCTATTTATTTTTTTATAGTTATGTTAGTATTAAGAACGTTATTTATATTTCAAATTTTTCTTTTTTTTCTGTACAGACGCGTGTACGCATGTAACATTATACTGAAAACCTTGCTTGAGAAGGTTTTGGGACGCTCGAAGGCTTTAATTTGC