pV1093 — PlasmidDB

Plasmid Information

Purpose

CaCas9/gRNA Solo entry plasmid for cloning guides - contains stuffer with BsmBI sites - integrates at ENO1

Depositor

Gerald Fink

Article

Vyas et al Sci Adv. 2015;1(3):e1500248.

Insert

CaCas9/sgRNA-BsmBI stuffer

Available Since

Oct. 17, 2018

Availability

Available

Sequence Information

Length

14,105 bp

Topology

circular

Molecule Type

other DNA

View on AddGene.org

Features

CDS 6

Label	Gene	Product	Position	Strand
Cas9	—	Cas9 (Csn1) endonuclease from the Strep…	2242..6346	Fwd
DNA Sequence ATGGATAAAAAGTATAGTATTGGTTTAGATATTGGTACTAACTCTGTGGGTTGGGCAGTTATCACTGACGAATATAAAGTTCCATCAAAGAAATTTAAGGTGTTAGGTAACACTGACAGACACTCAATAAAAAAGAATCTTATCGGTGCTCTTTTGTTCGACTCCGGTGAAACTGCCGAGGCTACACGTTTAAAAAGAACAGCAAGAAGAAGATATACCCGTAGAAAAAATAGAATATGTTATTTACAAGAAATCTTTTCTAATGAAATGGCTAAAGTTGATGATTCCTTTTTCCATAGATTGGAAGAGTCATTTTTGGTTGAAGAAGACAAAAAGCATGAGAGACATCCAATCTTTGGGAATATAGTTGATGAAGTGGCTTACCATGAAAAATATCCTACCATTTATCATTTAAGAAAGAAATTGGTAGATTCAACTGATAAAGCTGACCTTAGATTAATCTATTTAGCACTTGCCCATATGATTAAATTTAGAGGTCATTTTTTGATTGAAGGTGATTTGAACCCAGATAATTCTGACGTGGATAAATTATTTATTCAATTAGTCCAAACCTACAACCAATTATTTGAGGAAAATCCAATTAATGCTAGTGGTGTCGATGCCAAAGCTATATTATCAGCCAGATTATCAAAATCTAGACGTTTGGAAAATTTGATTGCCCAATTGCCAGGAGAAAAAAAGAATGGATTATTTGGAAACTTGATCGCATTATCATTGGGTTTGACACCAAATTTTAAATCTAATTTTGATTTAGCTGAAGATGCTAAATTACAATTATCAAAAGACACCTATGACGACGATTTGGACAATTTACTTGCTCAAATTGGTGATCAATATGCAGATTTGTTCTTAGCTGCTAAAAACTTATCTGATGCTATTTTGTTGTCTGATATTTTGAGAGTGAACACAGAAATAACCAAAGCTCCATTATCAGCATCTATGATCAAACGTTATGATGAACACCATCAGGATTTGACTTTATTGAAAGCTTTGGTGAGACAACAATTGCCAGAGAAGTATAAAGAAATCTTTTTCGATCAATCTAAAAACGGGTATGCAGGTTATATTGATGGGGGTGCCTCCCAAGAGGAATTTTACAAATTTATAAAACCTATTTTAGAAAAGATGGATGGGACTGAGGAACTTTTGGTCAAATTGAACAGAGAAGATTTGTTACGTAAACAGAGAACTTTTGATAATGGTAGTATACCTCACCAAATTCATTTGGGTGAGTTGCATGCAATTTTAAGAAGACAAGAAGATTTTTATCCATTTTTAAAAGATAATAGAGAAAAAATCGAGAAAATTTTAACCTTTAGAATTCCATACTATGTTGGGCCTTTGGCTAGAGGTAATTCAAGATTTGCCTGGATGACACGTAAATCAGAAGAAACTATTACCCCTTGGAATTTTGAAGAGGTTGTTGATAAAGGAGCATCAGCACAGAGTTTTATTGAAAGAATGACCAATTTCGATAAAAACTTACCAAATGAAAAAGTTTTACCAAAACATTCCTTGTTATACGAATATTTTACTGTTTACAATGAACTTACAAAGGTTAAATATGTTACTGAAGGTATGCGTAAGCCAGCCTTTTTATCTGGAGAACAGAAAAAGGCAATAGTTGATTTATTGTTTAAAACAAATAGAAAAGTTACTGTTAAACAATTAAAAGAAGATTACTTTAAGAAAATTGAATGTTTTGATTCAGTTGAAATCAGTGGTGTTGAAGACAGATTTAATGCTAGTTTAGGAACTTACCATGATTTACTTAAAATTATCAAAGATAAAGATTTCTTGGATAACGAAGAAAATGAAGACATTTTAGAAGACATTGTTTTAACCTTAACTTTATTCGAAGATAGAGAGATGATTGAAGAACGTTTGAAGACTTATGCACATTTGTTTGACGATAAAGTGATGAAACAGTTGAAAAGAAGACGTTATACTGGATGGGGTAGATTGTCTCGTAAATTGATCAATGGAATTAGAGATAAACAAAGTGGTAAAACTATCTTGGACTTTTTGAAATCTGACGGATTTGCTAATAGAAATTTCATGCAATTGATCCACGACGATAGTTTGACATTTAAAGAAGACATCCAAAAGGCCCAAGTGAGTGGGCAAGGTGATTCATTACATGAACATATTGCAAATTTAGCCGGATCTCCTGCTATTAAGAAAGGGATATTACAAACTGTTAAAGTTGTGGATGAATTAGTGAAAGTAATGGGAAGACATAAACCTGAAAACATTGTCATTGAGATGGCAAGAGAAAATCAAACTACACAAAAAGGACAGAAAAATAGTAGAGAACGTATGAAAAGAATAGAAGAGGGTATTAAAGAATTGGGTAGTCAAATATTGAAAGAACACCCAGTGGAAAATACCCAGTTGCAAAATGAAAAATTATATCTTTACTACCTTCAAAATGGACGTGATATGTATGTTGATCAGGAATTAGATATAAATAGACTTTCAGATTATGATGTAGATCATATAGTTCCACAATCTTTCTTGAAAGATGATTCCATAGACAATAAAGTATTAACTAGAAGTGATAAAAATAGAGGTAAAAGTGATAATGTCCCAAGTGAGGAAGTCGTCAAAAAGATGAAAAATTACTGGCGTCAACTTTTGAATGCTAAATTAATTACTCAAAGAAAATTTGATAATTTGACTAAAGCAGAAAGAGGTGGGCTTTCTGAATTAGATAAAGCCGGGTTCATTAAAAGACAATTGGTCGAAACTAGACAAATTACTAAACATGTTGCCCAAATTTTAGATTCCCGTATGAACACTAAGTATGACGAAAATGATAAGTTAATACGTGAGGTTAAAGTCATTACTTTAAAATCAAAACTTGTCTCTGATTTCAGAAAGGATTTCCAATTCTATAAAGTTAGAGAAATTAATAATTATCATCATGCTCATGATGCATATTTGAATGCTGTAGTTGGAACTGCTTTAATCAAGAAATACCCTAAATTAGAATCTGAATTTGTATATGGTGATTACAAAGTCTATGATGTTAGAAAGATGATTGCTAAATCAGAACAAGAAATTGGTAAAGCTACAGCTAAATACTTCTTTTACTCTAACATTATGAATTTCTTTAAAACAGAAATTACTTTGGCAAACGGTGAAATTAGAAAAAGACCTCTTATTGAAACAAATGGTGAGACTGGAGAGATAGTTTGGGACAAAGGGCGTGATTTCGCTACTGTTAGAAAAGTTTTATCAATGCCACAAGTTAACATTGTAAAGAAAACAGAGGTTCAAACTGGTGGTTTCTCAAAAGAAAGTATTTTGCCTAAAAGAAATAGTGATAAATTGATTGCCAGAAAAAAGGATTGGGATCCAAAGAAATATGGTGGTTTCGACTCACCAACCGTAGCCTATTCTGTTTTGGTTGTGGCAAAGGTTGAAAAGGGTAAAAGTAAAAAGCTTAAATCAGTAAAAGAACTTTTGGGTATTACAATAATGGAAAGAAGTTCCTTTGAAAAGAACCCTATTGATTTTTTGGAAGCTAAAGGTTATAAGGAAGTAAAGAAGGACTTAATAATCAAATTGCCTAAATATTCTTTATTTGAATTAGAAAATGGGAGAAAAAGAATGTTGGCTTCTGCTGGAGAATTGCAAAAGGGTAATGAATTAGCATTGCCTTCCAAATATGTTAACTTCTTGTATTTAGCTTCACACTATGAAAAGTTGAAAGGGTCACCAGAAGATAACGAGCAAAAACAATTATTTGTTGAACAACACAAACACTACTTAGATGAGATTATAGAACAAATTAGTGAATTCAGTAAAAGAGTGATATTAGCTGATGCAAATTTAGATAAAGTTTTGTCAGCCTATAACAAACATAGAGATAAGCCAATTAGAGAACAAGCAGAAAACATTATTCACTTATTTACCCTTACCAATTTAGGAGCACCTGCTGCTTTCAAGTATTTTGATACAACAATTGATCGTAAAAGATATACCTCAACAAAAGAAGTCTTAGACGCCACCTTAATTCATCAATCAATCACTGGATTGTATGAGACAAGAATTGATTTGTCTCAATTGGGTGGTGAT Protein Sequence MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD
SV40 NLS	—	nuclear localization signal of SV40 (si…	6358..6379	Fwd
DNA Sequence CCTAAGAAGAAAAGAAAAGTT Protein Sequence PKKKRKV
SV40 NLS	—	nuclear localization signal of SV40 (si…	6382..6403	Fwd
DNA Sequence CCAAAGAAAAAGCGTAAGGTG Protein Sequence PKKKRKV
SV40 NLS	—	nuclear localization signal of SV40 (si…	6406..6427	Fwd
DNA Sequence CCTAAGAAAAAGAGAAAGGTT Protein Sequence PKKKRKV
3xFLAG	—	three tandem FLAG(R) epitope tags, foll…	6427..6493	Fwd
DNA Sequence GACTACAAAGACCATGACGGTGATTATAAAGATCATGACATCGACTACAAGGATGACGATGACAAG Protein Sequence DYKDHDGDYKDHDIDYKDDDDK
AmpR	bla	beta-lactamase	13128..13989	Rev
DNA Sequence ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA Protein Sequence MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW

promoter 4

Label	Gene	Product	Position	Strand
T7 promoter	—	—	637..656	Fwd
DNA Sequence TAATACGACTCACTATAGG
T3 promoter	—	—	11928..11947	Rev
DNA Sequence AATTAACCCTCACTAAAGG
lac promoter	—	—	12015..12046	Rev
DNA Sequence TTTACACTTTATGCTCCCGGCTCGTATGTTG
AmpR promoter	bla	—	13989..14094	Rev
DNA Sequence CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT

protein_bind 2

Label	Gene	Product	Position	Strand
lac operator	—	—	11991..12008	Fwd
DNA Sequence TTGTTATCCGCTCACAA
CAP binding site	—	—	12060..12082	Fwd
DNA Sequence ATGAGTGAGCTAACTCACATTA

rep_origin 2

Label	Gene	Product	Position	Strand
M13 ori	—	—	14..470	Rev
DNA Sequence ACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTT
ori	—	—	12369..12958	Rev
DNA Sequence TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA

terminator 1

Label	Gene	Product	Position	Strand
CYC1 terminator	S. cerevisiae CYC1	—	6557..6805	Fwd
DNA Sequence TCATGTAATTAGTTATGTCACGCTTACATTCACGCCCTCCCCCCACATCCGCTCTAACCGAAAAGGAAGGAGTTAGACAACCTGAAGTCTAGGTCCCTATTTATTTTTTTATAGTTATGTTAGTATTAAGAACGTTATTTATATTTCAAATTTTTCTTTTTTTTCTGTACAGACGCGTGTACGCATGTAACATTATACTGAAAACCTTGCTTGAGAAGGTTTTGGGACGCTCGAAGGCTTTAATTTGC