YN2_1_IL50_HOlocus — PlasmidDB

Plasmid Information

Purpose

Expresses Cas9 and sgRNA cassettes for targeting the HO locus in yeast Saccharomyces cerevisiae

Depositor

Lisbeth Olsson

Article

Torello Pianale et al ACS Synth Biol. 2023 Aug 8. doi: 10.1021

Insert

Cas9

Mutation

Available Since

May 8, 2023

Availability

Available

Sequence Information

Length

9,432 bp

Topology

circular

Molecule Type

other DNA

View on AddGene.org

Features

CDS 5

Label	Gene	Product	Position	Strand
Cas9	—	Cas9 (Csn1) endonuclease from the Strep…	1182..5286	Fwd
DNA Sequence ATGGACAAGAAGTATTCTATCGGACTGGATATCGGGACTAATAGCGTCGGGTGGGCCGTGATCACTGACGAGTACAAGGTGCCCTCTAAGAAGTTCAAGGTGCTCGGGAACACCGACCGGCATTCCATCAAGAAAAATCTGATCGGAGCTCTCCTCTTTGATTCAGGGGAAACCGCTGAAGCAACCCGCCTCAAGCGGACTGCTAGACGGCGGTACACCAGGAGGAAGAACCGGATTTGTTACCTTCAAGAGATATTCTCCAACGAAATGGCAAAGGTCGACGACAGCTTCTTCCATAGGCTGGAAGAATCATTCCTCGTGGAAGAGGATAAGAAGCATGAACGGCATCCCATCTTCGGTAATATCGTCGACGAGGTGGCCTATCACGAGAAATACCCAACCATCTACCATCTTCGCAAAAAGCTGGTGGACTCAACCGACAAGGCAGACCTCCGGCTTATCTACCTGGCCCTGGCCCACATGATCAAGTTCAGAGGCCACTTCCTGATCGAGGGCGACCTCAATCCTGACAATAGCGATGTGGATAAACTGTTCATCCAGCTGGTGCAGACTTACAACCAGCTCTTTGAAGAGAACCCCATCAATGCAAGCGGAGTCGATGCCAAGGCCATTCTGTCAGCCCGGCTGTCAAAGAGCCGCAGACTTGAGAATCTTATCGCTCAGCTGCCGGGTGAAAAGAAAAATGGACTGTTCGGGAACCTGATTGCTCTTTCACTTGGGCTGACTCCCAATTTCAAGTCTAATTTCGACCTGGCAGAGGATGCCAAGCTGCAACTGTCCAAGGACACCTATGATGACGATCTCGACAACCTCCTGGCCCAGATCGGTGACCAATACGCCGACCTTTTCCTTGCTGCTAAGAATCTTTCTGACGCCATCCTGCTGTCTGACATTCTCCGCGTGAACACTGAAATCACCAAGGCCCCTCTTTCAGCTTCAATGATTAAGCGGTATGATGAGCACCACCAGGACCTGACCCTGCTTAAGGCACTCGTCCGGCAGCAGCTTCCGGAGAAGTACAAGGAAATCTTCTTTGACCAGTCAAAGAATGGATACGCCGGCTACATCGACGGAGGTGCCTCCCAAGAGGAATTTTATAAGTTTATCAAACCTATCCTTGAGAAGATGGACGGCACCGAAGAGCTCCTCGTGAAACTGAATCGGGAGGATCTGCTGCGGAAGCAGCGCACTTTCGACAATGGGAGCATTCCCCACCAGATCCATCTTGGGGAGCTTCACGCCATCCTTCGGCGCCAAGAGGACTTCTACCCCTTTCTTAAGGACAACAGGGAGAAGATTGAGAAAATTCTCACTTTCCGCATCCCCTACTACGTGGGACCCCTCGCCAGAGGAAATAGCCGGTTTGCTTGGATGACCAGAAAGTCAGAAGAAACTATCACTCCCTGGAACTTCGAAGAGGTGGTGGACAAGGGAGCCAGCGCTCAGTCATTCATCGAACGGATGACTAACTTCGATAAGAACCTCCCCAATGAGAAGGTCCTGCCGAAACATTCCCTGCTCTACGAGTACTTTACCGTGTACAACGAGCTGACCAAGGTGAAATATGTCACCGAAGGGATGAGGAAGCCCGCATTCCTGTCAGGCGAACAAAAGAAGGCAATTGTGGACCTTCTGTTCAAGACCAATAGAAAGGTGACCGTGAAGCAGCTGAAGGAGGACTATTTCAAGAAAATTGAATGCTTCGACTCTGTGGAGATTAGCGGGGTCGAAGATCGGTTCAACGCAAGCCTGGGTACTTACCATGATCTGCTTAAGATCATCAAGGACAAGGATTTTCTGGACAATGAGGAGAACGAGGACATCCTTGAGGACATTGTCCTGACTCTCACTCTGTTCGAGGACCGGGAAATGATCGAGGAGAGGCTTAAGACCTACGCCCATCTGTTCGACGATAAAGTGATGAAGCAACTTAAACGGAGAAGATATACCGGATGGGGACGCCTTAGCCGCAAACTCATCAACGGAATCCGGGACAAACAGAGCGGAAAGACCATTCTTGATTTCCTTAAGAGCGACGGATTCGCTAATCGCAACTTCATGCAACTTATCCATGATGATTCCCTGACCTTTAAGGAGGACATCCAGAAGGCCCAAGTGTCTGGACAAGGTGACTCACTGCACGAGCATATCGCAAATCTGGCTGGTTCACCCGCTATTAAGAAGGGTATTCTCCAGACCGTGAAAGTCGTGGACGAGCTGGTCAAGGTGATGGGTCGCCATAAACCAGAGAACATTGTCATCGAGATGGCCAGGGAAAACCAGACTACCCAGAAGGGACAGAAGAACAGCAGGGAGCGGATGAAAAGAATTGAGGAAGGGATTAAGGAGCTCGGGTCACAGATCCTTAAAGAGCACCCGGTGGAAAACACCCAGCTTCAGAATGAGAAGCTCTATCTGTACTACCTTCAAAATGGACGCGATATGTATGTGGACCAAGAGCTTGATATCAACAGGCTCTCAGACTACGACGTGGACCATATCGTCCCTCAGAGCTTCCTCAAAGACGACTCAATTGACAATAAGGTGCTGACTCGCTCAGACAAGAACCGGGGAAAGTCAGATAACGTGCCCTCAGAGGAAGTCGTGAAAAAGATGAAGAACTATTGGCGCCAGCTTCTGAACGCAAAGCTGATCACTCAGCGGAAGTTCGACAATCTCACTAAGGCTGAGAGGGGCGGACTGAGCGAACTGGACAAAGCAGGATTCATTAAACGGCAACTTGTGGAGACTCGGCAGATTACTAAACATGTCGCCCAAATCCTTGACTCACGCATGAATACCAAGTACGACGAAAACGACAAACTTATCCGCGAGGTGAAGGTGATTACCCTGAAGTCCAAGCTGGTCAGCGATTTCAGAAAGGACTTTCAATTCTACAAAGTGCGGGAGATCAATAACTATCATCATGCTCATGACGCATATCTGAATGCCGTGGTGGGAACCGCCCTGATCAAGAAGTACCCAAAGCTGGAAAGCGAGTTCGTGTACGGAGACTACAAGGTCTACGACGTGCGCAAGATGATTGCCAAATCTGAGCAGGAGATCGGAAAGGCCACCGCAAAGTACTTCTTCTACAGCAACATCATGAATTTCTTCAAGACCGAAATCACCCTTGCAAACGGTGAGATCCGGAAGAGGCCGCTCATCGAGACTAATGGGGAGACTGGCGAAATCGTGTGGGACAAGGGCAGAGATTTCGCTACCGTGCGCAAAGTGCTTTCTATGCCTCAAGTGAACATCGTGAAGAAAACCGAGGTGCAAACCGGAGGCTTTTCTAAGGAATCAATCCTCCCCAAGCGCAACTCCGACAAGCTCATTGCAAGGAAGAAGGATTGGGACCCTAAGAAGTACGGCGGATTCGATTCACCAACTGTGGCTTATTCTGTCCTGGTCGTGGCTAAGGTGGAAAAAGGAAAGTCTAAGAAGCTCAAGAGCGTGAAGGAACTGCTGGGTATCACCATTATGGAGCGCAGCTCCTTCGAGAAGAACCCAATTGACTTTCTCGAAGCCAAAGGTTACAAGGAAGTCAAGAAGGACCTTATCATCAAGCTCCCAAAGTATAGCCTGTTCGAACTGGAGAATGGGCGGAAGCGGATGCTCGCCTCCGCTGGCGAACTTCAGAAGGGTAATGAGCTGGCTCTCCCCTCCAAGTACGTGAATTTCCTCTACCTTGCAAGCCATTACGAGAAGCTGAAGGGGAGCCCCGAGGACAACGAGCAAAAGCAACTGTTTGTGGAGCAGCATAAGCATTATCTGGACGAGATCATTGAGCAGATTTCCGAGTTTTCTAAACGCGTCATTCTCGCTGATGCCAACCTCGATAAAGTCCTTAGCGCATACAATAAGCACAGAGACAAACCAATTCGGGAGCAGGCTGAGAATATCATCCACCTGTTCACCCTCACCAATCTTGGTGCCCCTGCCGCATTCAAGTACTTCGACACCACCATCGACCGGAAACGCTATACCTCCACCAAAGAAGTGCTGGACGCCACCCTCATCCACCAGAGCATCACCGGACTTTACGAAACTCGGATTGACCTCTCACAGCTCGGAGGGGAT Protein Sequence MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD
SV40 NLS	—	nuclear localization signal of SV40 (si…	5298..5319	Fwd
DNA Sequence CCAAAAAAGAAGAGAAAGGTA Protein Sequence PKKKRKV
SV40 NLS	—	nuclear localization signal of SV40 (si…	5322..5343	Fwd
DNA Sequence CCAAAAAAGAAGAGAAAGGTA Protein Sequence PKKKRKV
KanR	aph(3')-Ia	aminoglycoside phosphotransferase	6017..6827	Fwd
DNA Sequence ATGGGTAAGGAAAAGACTCACGTTTCGAGGCCGCGATTAAATTCCAACATGGATGCTGATTTATATGGGTATAAATGGGCTCGCGATAATGTCGGGCAATCAGGTGCGACAATCTATCGATTGTATGGGAAGCCCGATGCGCCAGAGTTGTTTCTGAAACATGGCAAAGGTAGCGTTGCCAATGATGTTACAGATGAGATGGTCAGACTAAACTGGCTGACGGAATTTATGCCTCTTCCGACCATCAAGCATTTTATCCGTACTCCTGATGATGCATGGTTACTCACCACTGCGATCCCCGGCAAAACAGCATTCCAGGTATTAGAAGAATATCCTGATTCAGGTGAAAATATTGTTGATGCGCTGGCAGTGTTCCTGCGCCGGTTGCATTCGATTCCTGTTTGTAATTGTCCTTTTAACAGCGATCGCGTATTTCGTCTGGCTCAGGCGCAATCACGAATGAATAACGGTTTGGTTGATGCGAGTGATTTTGATGACGAGCGTAATGGCTGGCCTGTTGAACAAGTCTGGAAAGAAATGCATAAGCTTTTGCCATTCTCACCGGATTCAGTCGTCACTCATGGTGATTTCTCACTTGATAACCTTATTTTTGACGAGGGGAAATTAATAGGTTGTATTGATGTTGGACGAGTCGGAATCGCAGACCGATACCAGGATCTTGCCATCCTATGGAACTGCCTCGGTGAGTTTTCTCCTTCATTACAGAAACGGCTTTTTCAAAAATATGGTATTGATAATCCTGATATGAATAAATTGCAGTTTCATTTGATGCTCGATGAGTTTTTCTAA Protein Sequence MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF
AmpR	bla	beta-lactamase	7647..8508	Rev
DNA Sequence ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCCCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA Protein Sequence MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW

promoter 1

Label	Gene	Product	Position	Strand
AmpR promoter	bla	—	8508..8613	Rev
DNA Sequence CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT

rep_origin 1

Label	Gene	Product	Position	Strand
ori	—	—	8715..9304	Fwd
DNA Sequence TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA

terminator 1

Label	Gene	Product	Position	Strand
TEF terminator	—	—	6831..7029	Fwd
DNA Sequence ACTGACAATAAAAAGATTCTTGTTTTCAAGAACTTGTCATTTGTATAGTTTTTTTATATTGTAGTTGTTCTATTTTAATCAAATGTTAGCGTGATTTATATTTTTTTTCGCCTCGACATCATCTGCCCAGATGCGAAGTTAAGTGCGCAGAAAGTAATATCATGCGTCAATCGTATGTGAATGCTGGTCGCTATACTG