pCRISPRa_VPR_yl — PlasmidDB

Plasmid Information

Purpose

CRISPR-dCas9-VPR vector for Yarrowia lipolytica, expressing dCas9-VPR and AvrII site for sgRNA insertion

Depositor

Ian Wheeldon

Article

Schwartz et al Biotechnol J. 2018 May 5:e1700584. doi:

Promoter

SCR1'-tRNA and UAS1B8-TEF(136)

Available Since

May 17, 2018

Availability

Available

Sequence Information

Length

13,377 bp

Topology

circular

Molecule Type

other DNA

View on AddGene.org

Features

CDS 6

Label	Gene	Product	Position	Strand
dCas9	—	catalytically dead mutant of the Cas9 e…	1357..5461	Fwd
DNA Sequence ATGGATAAGAAATACTCCATTGGCCTGGCCATCGGAACCAACTCCGTGGGTTGGGCCGTGATCACCGATGAGTACAAGGTGCCCTCTAAGAAATTCAAGGTCCTGGGCAACACCGACCGACACTCCATCAAGAAGAACCTGATCGGCGCTCTGCTCTTCGACTCTGGCGAGACCGCCGAGGCCACCCGACTGAAGCGAACCGCTCGAAGACGATACACCCGAAGAAAGAACCGAATCTGTTACCTGCAGGAGATCTTCTCTAACGAGATGGCCAAGGTGGACGACTCTTTCTTCCACCGACTGGAGGAGTCTTTCCTGGTGGAGGAGGACAAGAAGCACGAGCGACACCCCATCTTCGGCAACATCGTGGACGAGGTGGCCTACCACGAGAAGTACCCCACCATCTACCACCTGCGAAAGAAGCTGGTGGACTCTACCGACAAGGCCGACCTGCGACTGATCTACCTGGCCCTGGCCCACATGATCAAGTTCCGAGGCCACTTCCTGATCGAGGGCGACCTGAACCCCGACAACTCTGACGTGGACAAGCTGTTCATCCAGCTGGTGCAGACCTACAACCAGCTCTTCGAAGAGAACCCCATTAACGCTTCTGGCGTGGATGCTAAGGCCATCCTGTCTGCCCGACTGTCTAAGTCTCGACGACTCGAGAACCTGATTGCTCAGCTCCCCGGAGAGAAGAAGAACGGTCTGTTCGGAAACCTGATTGCTCTGTCCCTGGGTCTCACCCCTAACTTCAAGTCCAACTTCGATCTGGCTGAGGACGCTAAGCTGCAGCTGTCTAAGGACACCTACGACGATGACCTGGATAACCTGCTCGCCCAGATTGGCGACCAGTACGCCGACCTGTTCCTGGCCGCCAAGAACCTGTCTGACGCCATCCTGCTGTCTGACATCCTGCGAGTGAACACCGAGATCACCAAGGCCCCCCTGTCTGCCTCCATGATTAAGCGATACGATGAGCACCACCAGGATCTGACCCTCCTCAAGGCTCTGGTCCGACAGCAGCTGCCCGAGAAGTACAAGGAGATTTTCTTCGACCAGTCTAAGAACGGCTACGCCGGCTACATCGACGGCGGCGCCTCTCAGGAGGAGTTCTACAAGTTCATTAAGCCCATCCTGGAGAAGATGGACGGAACCGAGGAACTGCTCGTGAAGCTGAACCGAGAGGACCTCCTGCGAAAGCAGCGAACCTTCGACAACGGCTCTATCCCCCACCAGATCCACCTGGGCGAGCTGCACGCCATCCTGCGACGACAGGAGGACTTCTACCCCTTCCTGAAGGACAACCGAGAGAAGATCGAGAAGATCCTGACCTTCCGAATCCCCTACTACGTGGGACCCCTGGCCCGAGGAAACTCTCGATTCGCTTGGATGACCCGAAAGTCTGAGGAGACCATTACCCCCTGGAACTTCGAGGAGGTGGTGGATAAGGGCGCCTCTGCTCAGTCTTTCATCGAGCGAATGACCAACTTCGACAAGAACCTCCCCAACGAGAAGGTCCTGCCCAAGCACTCTCTGCTCTACGAGTACTTCACCGTCTACAACGAGCTCACCAAGGTCAAGTACGTGACCGAGGGAATGCGAAAGCCCGCTTTCCTGTCTGGAGAGCAGAAGAAGGCTATTGTGGATCTGCTCTTCAAGACTAACCGAAAGGTCACCGTCAAGCAGCTGAAGGAGGATTACTTCAAGAAGATTGAGTGTTTCGATTCTGTCGAGATCTCCGGCGTCGAGGACCGATTCAACGCCTCTCTGGGTACCTACCACGACCTGCTGAAGATTATCAAGGACAAGGATTTCCTGGATAACGAGGAGAACGAGGATATTCTCGAGGACATTGTCCTGACCCTCACCCTGTTCGAGGATCGAGAGATGATTGAGGAGCGACTCAAGACCTACGCTCACCTGTTCGACGACAAGGTGATGAAGCAGCTGAAGCGACGACGATACACCGGCTGGGGCCGACTGTCTCGAAAGCTGATCAACGGCATCCGAGACAAGCAGTCTGGCAAGACCATCCTGGACTTCCTGAAGTCTGACGGCTTCGCCAACCGAAACTTCATGCAGCTGATCCACGACGACTCTCTGACCTTCAAGGAGGACATCCAGAAGGCCCAGGTGTCTGGCCAGGGCGACTCTCTGCACGAGCACATCGCCAACCTGGCCGGCTCTCCCGCCATTAAGAAAGGTATCCTGCAGACCGTCAAGGTGGTCGATGAGCTCGTCAAGGTGATGGGCCGACACAAGCCCGAGAACATTGTCATTGAGATGGCTCGAGAGAACCAGACTACTCAGAAGGGTCAGAAAAACTCCCGAGAGCGAATGAAGCGAATTGAGGAAGGTATTAAGGAGCTGGGATCCCAGATTCTCAAGGAGCATCCCGTGGAGAACACTCAGCTCCAGAACGAGAAGCTGTACCTGTACTATCTGCAGAACGGTCGAGACATGTACGTCGACCAGGAGCTGGATATCAACCGACTCTCCGACTACGATGTGGACGCCATTGTGCCCCAGTCCTTCCTGAAGGACGATTCTATCGATAACAAGGTGCTGACCCGATCCGACAAGAACCGAGGCAAGTCTGACAACGTGCCCTCCGAGGAGGTGGTCAAGAAGATGAAGAACTACTGGCGACAGCTGCTGAACGCCAAGCTGATTACCCAGCGAAAGTTCGACAACCTGACCAAGGCCGAGCGAGGCGGCCTGTCTGAGCTGGACAAGGCCGGCTTCATCAAGCGACAGCTGGTGGAGACCCGACAGATCACCAAGCACGTGGCCCAGATCCTGGACTCTCGAATGAACACCAAGTACGACGAGAACGACAAGCTGATCCGAGAGGTGAAGGTGATCACCCTGAAGTCTAAGCTGGTGTCTGACTTCCGAAAGGACTTCCAGTTCTACAAGGTGCGAGAGATTAACAACTACCACCACGCCCACGATGCCTACCTGAACGCTGTCGTGGGCACCGCCCTCATCAAGAAGTATCCCAAGCTGGAGTCCGAGTTCGTCTACGGCGACTACAAGGTCTACGATGTGCGAAAAATGATTGCCAAGTCCGAGCAGGAGATTGGCAAGGCTACCGCCAAGTACTTCTTCTACTCCAACATTATGAACTTCTTCAAGACCGAGATTACCCTGGCTAACGGCGAGATTCGAAAGCGACCCCTCATTGAGACCAACGGAGAGACCGGTGAGATCGTGTGGGACAAGGGACGAGACTTCGCCACCGTGCGAAAGGTGCTGTCTATGCCCCAGGTGAACATCGTGAAGAAGACCGAGGTGCAGACCGGAGGTTTCTCTAAGGAGTCCATCCTGCCCAAGCGAAACTCTGACAAGCTGATCGCCCGAAAGAAGGACTGGGACCCCAAGAAGTACGGAGGTTTCGACTCTCCCACCGTGGCTTACTCTGTGCTGGTGGTGGCCAAGGTGGAGAAGGGCAAGTCTAAGAAGCTGAAGTCTGTGAAGGAGCTGCTGGGCATTACCATCATGGAGCGATCTTCTTTCGAGAAGAACCCCATTGACTTCCTGGAGGCCAAGGGATACAAGGAGGTGAAGAAAGATCTGATTATCAAGCTCCCCAAGTACTCTCTGTTCGAGCTGGAGAACGGACGAAAGCGAATGCTGGCCTCTGCCGGCGAGCTGCAGAAGGGAAACGAGCTGGCCCTGCCCTCCAAGTACGTCAACTTCCTGTACCTCGCCTCCCATTACGAGAAGCTGAAGGGCTCTCCCGAGGATAACGAGCAGAAGCAGCTCTTCGTGGAGCAGCATAAGCACTACCTGGACGAGATCATCGAGCAGATCTCTGAGTTCTCTAAGCGAGTGATCCTGGCTGACGCCAACCTGGATAAGGTGCTGTCTGCTTACAACAAGCACCGAGACAAGCCCATTCGAGAGCAGGCTGAGAACATCATTCACCTGTTCACCCTGACCAACCTGGGAGCCCCCGCTGCCTTCAAGTACTTCGACACCACCATCGACCGAAAGCGATACACCTCTACCAAGGAGGTGCTGGACGCCACCCTGATCCACCAGTCTATCACCGGCCTGTACGAGACCCGAATCGACCTGTCTCAGCTGGGCGGCGAC Protein Sequence MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD
SV40 NLS	—	nuclear localization signal of SV40 (si…	5473..5494	Fwd
DNA Sequence CCCAAGAAGAAGCGAAAGGTG Protein Sequence PKKKRKV
VP64	—	tetrameric repeat of the minimal activa…	5506..5656	Fwd
DNA Sequence GACGCTCTCGATGACTTTGATCTGGACATGCTTGGCTCCGATGCTCTTGACGATTTCGACCTCGACATGCTTGGATCTGACGCTCTGGATGACTTCGACCTTGATATGCTCGGATCTGATGCCCTTGACGATTTCGATCTGGACATGCTG Protein Sequence DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDML
SV40 NLS	—	nuclear localization signal of SV40 (si…	5674..5695	Fwd
DNA Sequence CCCAAGAAGAAGCGAAAGGTG Protein Sequence PKKKRKV
Rta AD	—	transcriptional activation domain from …	6499..7069	Fwd
DNA Sequence CGAGACTCTCGAGAGGGCATGTTCCTGCCCAAGCCCGAGGCCGGCTCTGCCATCTCTGACGTGTTCGAGGGCCGAGAGGTGTGCCAGCCCAAGCGAATCCGACCCTTCCACCCACCCGGCTCTCCTTGGGCTAACCGACCCCTGCCTGCTTCTCTGGCACCAACCCCCACCGGCCCTGTGCACGAGCCAGTGGGCTCTCTGACCCCTGCTCCAGTGCCCCAGCCTCTGGACCCTGCCCCCGCTGTGACCCCCGAGGCCTCTCACCTGCTGGAGGACCCCGACGAGGAGACCTCTCAGGCCGTGAAGGCCCTGCGAGAGATGGCCGACACCGTGATCCCCCAGAAGGAGGAGGCCGCCATCTGCGGCCAGATGGACCTGTCTCACCCTCCACCCCGAGGCCACCTGGACGAGCTGACCACCACCCTGGAGTCTATGACCGAGGACCTGAACCTGGACTCTCCCCTGACCCCCGAGCTGAACGAGATCCTGGACACCTTCCTGAACGACGAGTGCCTGCTGCACGCCATGCACATCTCTACCGGCCTGTCTATCTTCGACACCTCTCTGTTC Protein Sequence RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWANRPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKALREMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEILDTFLNDECLLHAMHISTGLSIFDTSLF
AmpR	bla	beta-lactamase	8506..9367	Rev
DNA Sequence ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA Protein Sequence MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW

promoter 2

Label	Gene	Product	Position	Strand
lac promoter	—	—	7393..7424	Rev
DNA Sequence TTTACACTTTATGCTTCCGGCTCGTATGTTG
AmpR promoter	bla	—	9367..9472	Rev
DNA Sequence CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT

protein_bind 2

Label	Gene	Product	Position	Strand
lac operator	—	—	7369..7386	Fwd
DNA Sequence TTGTTATCCGCTCACAA
CAP binding site	—	—	7438..7460	Fwd
DNA Sequence ATGAGTGAGCTAACTCACATTA

rep_origin 1

Label	Gene	Product	Position	Strand
ori	—	—	7747..8336	Rev
DNA Sequence TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA

terminator 1

Label	Gene	Product	Position	Strand
CYC1 terminator	S. cerevisiae CYC1	—	7079..7327	Fwd
DNA Sequence TCATGTAATTAGTTATGTCACGCTTACATTCACGCCCTCCCTCCACATCCGCTCTAACCGAAAAGGAAGGAGTTAGACAACCTGAAGTCTAGGTCCCTATTTATTTTTTTATAGTTATGTTAGTATTAAGAACGTTATTTATATTTCAAATTTTTCTTTTTTTTCTGTACAGACGCGTGTACGCATGTAACATTATACTGAAAACCTTGCTTGAGAAGGTTTTGGGACGCTCGAAGGCTTTAATTTGC