HCP1

166103 Saccharomyces cerevisiae Yeast Expression

Plasmid Information

Purpose
This plasmid encodes a Cas9 protein as well as a sgRNA that targets Msn2, which will create a double strand break and allow C-terminus tagging via homologous recombination.
Depositor
Matthias Heinemann
Insert
Msn2-sgRNA#27 (MSN2 Budding Yeast)
Available Since
April 14, 2021
Availability
Available

Sequence Information

Length
9,440 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… 872..4976 Fwd
DNA Sequence
ATGGACAAGAAGTATTCTATCGGACTGGATATCGGGACTAATAGCGTCGGGTGGGCCGTGATCACTGACGAGTACAAGGTGCCCTCTAAGAAGTTCAAGGTGCTCGGGAACACCGACCGGCATTCCATCAAGAAAAATCTGATCGGAGCTCTCCTCTTTGATTCAGGGGAAACCGCTGAAGCAACCCGCCTCAAGCGGACTGCTAGACGGCGGTACACCAGGAGGAAGAACCGGATTTGTTACCTTCAAGAGATATTCTCCAACGAAATGGCAAAGGTCGACGACAGCTTCTTCCATAGGCTGGAAGAATCATTCCTCGTGGAAGAGGATAAGAAGCATGAACGGCATCCCATCTTCGGTAATATCGTCGACGAGGTGGCCTATCACGAGAAATACCCAACCATCTACCATCTTCGCAAAAAGCTGGTGGACTCAACCGACAAGGCAGACCTCCGGCTTATCTACCTGGCCCTGGCCCACATGATCAAGTTCAGAGGCCACTTCCTGATCGAGGGCGACCTCAATCCTGACAATAGCGATGTGGATAAACTGTTCATCCAGCTGGTGCAGACTTACAACCAGCTCTTTGAAGAGAACCCCATCAATGCAAGCGGAGTCGATGCCAAGGCCATTCTGTCAGCCCGGCTGTCAAAGAGCCGCAGACTTGAGAATCTTATCGCTCAGCTGCCGGGTGAAAAGAAAAATGGACTGTTCGGGAACCTGATTGCTCTTTCACTTGGGCTGACTCCCAATTTCAAGTCTAATTTCGACCTGGCAGAGGATGCCAAGCTGCAACTGTCCAAGGACACCTATGATGACGATCTCGACAACCTCCTGGCCCAGATCGGTGACCAATACGCCGACCTTTTCCTTGCTGCTAAGAATCTTTCTGACGCCATCCTGCTGTCTGACATTCTCCGCGTGAACACTGAAATCACCAAGGCCCCTCTTTCAGCTTCAATGATTAAGCGGTATGATGAGCACCACCAGGACCTGACCCTGCTTAAGGCACTCGTCCGGCAGCAGCTTCCGGAGAAGTACAAGGAAATCTTCTTTGACCAGTCAAAGAATGGATACGCCGGCTACATCGACGGAGGTGCCTCCCAAGAGGAATTTTATAAGTTTATCAAACCTATCCTTGAGAAGATGGACGGCACCGAAGAGCTCCTCGTGAAACTGAATCGGGAGGATCTGCTGCGGAAGCAGCGCACTTTCGACAATGGGAGCATTCCCCACCAGATCCATCTTGGGGAGCTTCACGCCATCCTTCGGCGCCAAGAGGACTTCTACCCCTTTCTTAAGGACAACAGGGAGAAGATTGAGAAAATTCTCACTTTCCGCATCCCCTACTACGTGGGACCCCTCGCCAGAGGAAATAGCCGGTTTGCTTGGATGACCAGAAAGTCAGAAGAAACTATCACTCCCTGGAACTTCGAAGAGGTGGTGGACAAGGGAGCCAGCGCTCAGTCATTCATCGAACGGATGACTAACTTCGATAAGAACCTCCCCAATGAGAAGGTCCTGCCGAAACATTCCCTGCTCTACGAGTACTTTACCGTGTACAACGAGCTGACCAAGGTGAAATATGTCACCGAAGGGATGAGGAAGCCCGCATTCCTGTCAGGCGAACAAAAGAAGGCAATTGTGGACCTTCTGTTCAAGACCAATAGAAAGGTGACCGTGAAGCAGCTGAAGGAGGACTATTTCAAGAAAATTGAATGCTTCGACTCTGTGGAGATTAGCGGGGTCGAAGATCGGTTCAACGCAAGCCTGGGTACTTACCATGATCTGCTTAAGATCATCAAGGACAAGGATTTTCTGGACAATGAGGAGAACGAGGACATCCTTGAGGACATTGTCCTGACTCTCACTCTGTTCGAGGACCGGGAAATGATCGAGGAGAGGCTTAAGACCTACGCCCATCTGTTCGACGATAAAGTGATGAAGCAACTTAAACGGAGAAGATATACCGGATGGGGACGCCTTAGCCGCAAACTCATCAACGGAATCCGGGACAAACAGAGCGGAAAGACCATTCTTGATTTCCTTAAGAGCGACGGATTCGCTAATCGCAACTTCATGCAACTTATCCATGATGATTCCCTGACCTTTAAGGAGGACATCCAGAAGGCCCAAGTGTCTGGACAAGGTGACTCACTGCACGAGCATATCGCAAATCTGGCTGGTTCACCCGCTATTAAGAAGGGTATTCTCCAGACCGTGAAAGTCGTGGACGAGCTGGTCAAGGTGATGGGTCGCCATAAACCAGAGAACATTGTCATCGAGATGGCCAGGGAAAACCAGACTACCCAGAAGGGACAGAAGAACAGCAGGGAGCGGATGAAAAGAATTGAGGAAGGGATTAAGGAGCTCGGGTCACAGATCCTTAAAGAGCACCCGGTGGAAAACACCCAGCTTCAGAATGAGAAGCTCTATCTGTACTACCTTCAAAATGGACGCGATATGTATGTGGACCAAGAGCTTGATATCAACAGGCTCTCAGACTACGACGTGGACCATATCGTCCCTCAGAGCTTCCTCAAAGACGACTCAATTGACAATAAGGTGCTGACTCGCTCAGACAAGAACCGGGGAAAGTCAGATAACGTGCCCTCAGAGGAAGTCGTGAAAAAGATGAAGAACTATTGGCGCCAGCTTCTGAACGCAAAGCTGATCACTCAGCGGAAGTTCGACAATCTCACTAAGGCTGAGAGGGGCGGACTGAGCGAACTGGACAAAGCAGGATTCATTAAACGGCAACTTGTGGAGACTCGGCAGATTACTAAACATGTCGCCCAAATCCTTGACTCACGCATGAATACCAAGTACGACGAAAACGACAAACTTATCCGCGAGGTGAAGGTGATTACCCTGAAGTCCAAGCTGGTCAGCGATTTCAGAAAGGACTTTCAATTCTACAAAGTGCGGGAGATCAATAACTATCATCATGCTCATGACGCATATCTGAATGCCGTGGTGGGAACCGCCCTGATCAAGAAGTACCCAAAGCTGGAAAGCGAGTTCGTGTACGGAGACTACAAGGTCTACGACGTGCGCAAGATGATTGCCAAATCTGAGCAGGAGATCGGAAAGGCCACCGCAAAGTACTTCTTCTACAGCAACATCATGAATTTCTTCAAGACCGAAATCACCCTTGCAAACGGTGAGATCCGGAAGAGGCCGCTCATCGAGACTAATGGGGAGACTGGCGAAATCGTGTGGGACAAGGGCAGAGATTTCGCTACCGTGCGCAAAGTGCTTTCTATGCCTCAAGTGAACATCGTGAAGAAAACCGAGGTGCAAACCGGAGGCTTTTCTAAGGAATCAATCCTCCCCAAGCGCAACTCCGACAAGCTCATTGCAAGGAAGAAGGATTGGGACCCTAAGAAGTACGGCGGATTCGATTCACCAACTGTGGCTTATTCTGTCCTGGTCGTGGCTAAGGTGGAAAAAGGAAAGTCTAAGAAGCTCAAGAGCGTGAAGGAACTGCTGGGTATCACCATTATGGAGCGCAGCTCCTTCGAGAAGAACCCAATTGACTTTCTCGAAGCCAAAGGTTACAAGGAAGTCAAGAAGGACCTTATCATCAAGCTCCCAAAGTATAGCCTGTTCGAACTGGAGAATGGGCGGAAGCGGATGCTCGCCTCCGCTGGCGAACTTCAGAAGGGTAATGAGCTGGCTCTCCCCTCCAAGTACGTGAATTTCCTCTACCTTGCAAGCCATTACGAGAAGCTGAAGGGGAGCCCCGAGGACAACGAGCAAAAGCAACTGTTTGTGGAGCAGCATAAGCATTATCTGGACGAGATCATTGAGCAGATTTCCGAGTTTTCTAAACGCGTCATTCTCGCTGATGCCAACCTCGATAAAGTCCTTAGCGCATACAATAAGCACAGAGACAAACCAATTCGGGAGCAGGCTGAGAATATCATCCACCTGTTCACCCTCACCAATCTTGGTGCCCCTGCCGCATTCAAGTACTTCGACACCACCATCGACCGGAAACGCTATACCTCCACCAAAGAAGTGCTGGACGCCACCCTCATCCACCAGAGCATCACCGGACTTTACGAAACTCGGATTGACCTCTCACAGCTCGGAGGGGAT
Protein Sequence
MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD
SV40 NLS — nuclear localization signal of SV40 (si… 4988..5009 Fwd
DNA Sequence
CCAAAAAAGAAGAGAAAGGTA
Protein Sequence
PKKKRKV
SV40 NLS — nuclear localization signal of SV40 (si… 5012..5033 Fwd
DNA Sequence
CCAAAAAAGAAGAGAAAGGTA
Protein Sequence
PKKKRKV
KanR aph(3')-Ia aminoglycoside phosphotransferase 6248..7058 Fwd
DNA Sequence
ATGGGTAAGGAAAAGACTCACGTTTCGAGGCCGCGATTAAATTCCAACATGGATGCTGATTTATATGGGTATAAATGGGCTCGCGATAATGTCGGGCAATCAGGTGCGACAATCTATCGATTGTATGGGAAGCCCGATGCGCCAGAGTTGTTTCTGAAACATGGCAAAGGTAGCGTTGCCAATGATGTTACAGATGAGATGGTCAGACTAAACTGGCTGACGGAATTTATGCCTCTTCCGACCATCAAGCATTTTATCCGTACTCCTGATGATGCATGGTTACTCACCACTGCGATCCCCGGCAAAACAGCATTCCAGGTATTAGAAGAATATCCTGATTCAGGTGAAAATATTGTTGATGCGCTGGCAGTGTTCCTGCGCCGGTTGCATTCGATTCCTGTTTGTAATTGTCCTTTTAACAGCGATCGCGTATTTCGTCTGGCTCAGGCGCAATCACGAATGAATAACGGTTTGGTTGATGCGAGTGATTTTGATGACGAGCGTAATGGCTGGCCTGTTGAACAAGTCTGGAAAGAAATGCATAAGCTTTTGCCATTCTCACCGGATTCAGTCGTCACTCATGGTGATTTCTCACTTGATAACCTTATTTTTGACGAGGGGAAATTAATAGGTTGTATTGATGTTGGACGAGTCGGAATCGCAGACCGATACCAGGATCTTGCCATCCTATGGAACTGCCTCGGTGAGTTTTCTCCTTCATTACAGAAACGGCTTTTTCAAAAATATGGTATTGATAATCCTGATATGAATAAATTGCAGTTTCATTTGATGCTCGATGAGTTTTTCTAA
Protein Sequence
MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF
KanR aph(3')-Ia aminoglycoside phosphotransferase 7811..8627 Rev
DNA Sequence
ATGAGCCATATTCAACGGGAAACGTCTTGCTCCAGGCCGCGATTAAATTCCAACATGGATGCTGATTTATATGGGTATAAATGGGCTCGCGATAATGTCGGGCAATCAGGTGCGACAATCTATCGATTGTATGGGAAGCCCGATGCGCCAGAGTTGTTTCTGAAACATGGCAAAGGTAGCGTTGCCAATGATGTTACAGATGAGATGGTCAGACTAAACTGGCTGACGGAATTTATGCCTCTTCCGACCATCAAGCATTTTATCCGTACTCCTGATGATGCATGGTTACTCACCACTGCGATCCCCGGGAAAACAGCATTCCAGGTATTAGAAGAATATCCTGATTCAGGTGAAAATATTGTTGATGCGCTGGCAGTGTTCCTGCGCCGGTTGCATTCGATTCCTGTTTGTAATTGTCCTTTTAACAGCGATCGCGTATTTCGCCTCGCTCAGGCGCAATCACGAATGAATAACGGTTTGGTTGATGCGAGTGATTTTGATGACGAGCGTAATGGCTGGCCTGTTGAACAAGTCTGGAAAGAAATGCATAAGCTTTTGCCATTCTCACCGGATTCAGTCGTCACTCATGGTGATTTCTCACTTGATAACCTTATTTTTGACGAGGGGAAATTAATAGGTTGTATTGATGTTGGACGAGTCGGAATCGCAGACCGATACCAGGATCTTGCCATCCTATGGAACTGCCTCGGTGAGTTTTCTCCTTCATTACAGAAACGGCTTTTTCAAAAATATGGTATTGATAATCCTGATATGAATAAATTGCAGTTTCATTTGATGCTCGATGAGTTTTTCTAA
Protein Sequence
MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF
rep_origin 1
Label Gene Product Position Strand
ori — — 8851..9440 Fwd
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 1
Label Gene Product Position Strand
TEF terminator — — 7062..7260 Fwd
DNA Sequence
ACTGACAATAAAAAGATTCTTGTTTTCAAGAACTTGTCATTTGTATAGTTTTTTTATATTGTAGTTGTTCTATTTTAATCAAATGTTAGCGTGATTTATATTTTTTTTCGCCTCGACATCATCTGCCCAGATGCGAAGTTAAGTGCGCAGAAAGTAATATCATGCGTCAATCGTATGTGAATGCTGGTCGCTATACTG