pV1326

111435 Synthetic Yeast Expression

Plasmid Information

Purpose
S. cerevisiae and C. glabrata Solo CRISPR vector, marked with URA3 and NatR
Depositor
Gerald Fink
Article
Vyas et al mSphere. 2018 Apr 25;3(2). pii: 3/2/e001
Insert
CaCas9/sgRNA-BsmBI stuffer
Available Since
Aug. 29, 2018
Availability
Available

Sequence Information

Length
14,517 bp
Topology
circular
Molecule Type
other DNA

Features

CDS 7
Label Gene Product Position Strand
URA3 S. cerevisiae URA3 orotidine-5'-phosphate decarboxylase, r… 416..1220 Fwd
DNA Sequence
ATGTCGAAAGCTACATATAAGGAACGTGCTGCTACTCATCCTAGTCCTGTTGCTGCCAAGCTATTTAATATCATGCACGAAAAGCAAACAAACTTGTGTGCTTCATTGGATGTTCGTACCACCAAGGAATTACTGGAGTTAGTTGAAGCATTAGGTCCCAAAATTTGTTTACTAAAAACACATGTGGATATCTTGACTGATTTTTCCATGGAGGGCACAGTTAAGCCGCTAAAGGCATTATCCGCCAAGTACAATTTTTTACTCTTCGAAGACAGAAAATTTGCTGACATTGGTAATACAGTCAAATTGCAGTACTCTGCGGGTGTATACAGAATAGCAGAATGGGCAGACATTACGAATGCACACGGTGTGGTGGGCCCAGGTATTGTTAGCGGTTTGAAGCAGGCGGCAGAAGAAGTAACAAAGGAACCTAGAGGCCTTTTGATGTTAGCAGAATTGTCATGCAAGGGCTCCCTATCTACTGGAGAATATACTAAGGGTACTGTTGACATTGCGAAGAGCGACAAAGATTTTGTTATCGGCTTTATTGCTCAAAGAGACATGGGTGGAAGAGATGAAGGTTACGATTGGTTGATTATGACACCCGGTGTGGGTTTAGATGACAAGGGTGATGCATTGGGTCAACAGTATAGAACCGTGGATGATGTGGTCTCTACAGGATCTGACATTATTATTGTTGGAAGAGGACTATTTGCAAAGGGAAGGGATGCTAAGGTAGAGGGTGAACGTTACAGAAAAGCAGGCTGGGAAGCATATTTGAGAAGATGCGGCCAGCAAAACTAA
Protein Sequence
MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELLELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQYSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGEYTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDDVVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN
Cas9 Cas9 (Csn1) endonuclease from the Strep… 3564..7668 Fwd
DNA Sequence
ATGGATAAAAAGTATAGTATTGGTTTAGATATTGGTACTAACTCTGTGGGTTGGGCAGTTATCACTGACGAATATAAAGTTCCATCAAAGAAATTTAAGGTGTTAGGTAACACTGACAGACACTCAATAAAAAAGAATCTTATCGGTGCTCTTTTGTTCGACTCCGGTGAAACTGCCGAGGCTACACGTTTAAAAAGAACAGCAAGAAGAAGATATACCCGTAGAAAAAATAGAATATGTTATTTACAAGAAATCTTTTCTAATGAAATGGCTAAAGTTGATGATTCCTTTTTCCATAGATTGGAAGAGTCATTTTTGGTTGAAGAAGACAAAAAGCATGAGAGACATCCAATCTTTGGGAATATAGTTGATGAAGTGGCTTACCATGAAAAATATCCTACCATTTATCATTTAAGAAAGAAATTGGTAGATTCAACTGATAAAGCTGACCTTAGATTAATCTATTTAGCACTTGCCCATATGATTAAATTTAGAGGTCATTTTTTGATTGAAGGTGATTTGAACCCAGATAATTCTGACGTGGATAAATTATTTATTCAATTAGTCCAAACCTACAACCAATTATTTGAGGAAAATCCAATTAATGCTAGTGGTGTCGATGCCAAAGCTATATTATCAGCCAGATTATCAAAATCTAGACGTTTGGAAAATTTGATTGCCCAATTGCCAGGAGAAAAAAAGAATGGATTATTTGGAAACTTGATCGCATTATCATTGGGTTTGACACCAAATTTTAAATCTAATTTTGATTTAGCTGAAGATGCTAAATTACAATTATCAAAAGACACCTATGACGACGATTTGGACAATTTACTTGCTCAAATTGGTGATCAATATGCAGATTTGTTCTTAGCTGCTAAAAACTTATCTGATGCTATTTTGTTGTCTGATATTTTGAGAGTGAACACAGAAATAACCAAAGCTCCATTATCAGCATCTATGATCAAACGTTATGATGAACACCATCAGGATTTGACTTTATTGAAAGCTTTGGTGAGACAACAATTGCCAGAGAAGTATAAAGAAATCTTTTTCGATCAATCTAAAAACGGGTATGCAGGTTATATTGATGGGGGTGCCTCCCAAGAGGAATTTTACAAATTTATAAAACCTATTTTAGAAAAGATGGATGGGACTGAGGAACTTTTGGTCAAATTGAACAGAGAAGATTTGTTACGTAAACAGAGAACTTTTGATAATGGTAGTATACCTCACCAAATTCATTTGGGTGAGTTGCATGCAATTTTAAGAAGACAAGAAGATTTTTATCCATTTTTAAAAGATAATAGAGAAAAAATCGAGAAAATTTTAACCTTTAGAATTCCATACTATGTTGGGCCTTTGGCTAGAGGTAATTCAAGATTTGCCTGGATGACACGTAAATCAGAAGAAACTATTACCCCTTGGAATTTTGAAGAGGTTGTTGATAAAGGAGCATCAGCACAGAGTTTTATTGAAAGAATGACCAATTTCGATAAAAACTTACCAAATGAAAAAGTTTTACCAAAACATTCCTTGTTATACGAATATTTTACTGTTTACAATGAACTTACAAAGGTTAAATATGTTACTGAAGGTATGCGTAAGCCAGCCTTTTTATCTGGAGAACAGAAAAAGGCAATAGTTGATTTATTGTTTAAAACAAATAGAAAAGTTACTGTTAAACAATTAAAAGAAGATTACTTTAAGAAAATTGAATGTTTTGATTCAGTTGAAATCAGTGGTGTTGAAGACAGATTTAATGCTAGTTTAGGAACTTACCATGATTTACTTAAAATTATCAAAGATAAAGATTTCTTGGATAACGAAGAAAATGAAGACATTTTAGAAGACATTGTTTTAACCTTAACTTTATTCGAAGATAGAGAGATGATTGAAGAACGTTTGAAGACTTATGCACATTTGTTTGACGATAAAGTGATGAAACAGTTGAAAAGAAGACGTTATACTGGATGGGGTAGATTGTCTCGTAAATTGATCAATGGAATTAGAGATAAACAAAGTGGTAAAACTATCTTGGACTTTTTGAAATCTGACGGATTTGCTAATAGAAATTTCATGCAATTGATCCACGACGATAGTTTGACATTTAAAGAAGACATCCAAAAGGCCCAAGTGAGTGGGCAAGGTGATTCATTACATGAACATATTGCAAATTTAGCCGGATCTCCTGCTATTAAGAAAGGGATATTACAAACTGTTAAAGTTGTGGATGAATTAGTGAAAGTAATGGGAAGACATAAACCTGAAAACATTGTCATTGAGATGGCAAGAGAAAATCAAACTACACAAAAAGGACAGAAAAATAGTAGAGAACGTATGAAAAGAATAGAAGAGGGTATTAAAGAATTGGGTAGTCAAATATTGAAAGAACACCCAGTGGAAAATACCCAGTTGCAAAATGAAAAATTATATCTTTACTACCTTCAAAATGGACGTGATATGTATGTTGATCAGGAATTAGATATAAATAGACTTTCAGATTATGATGTAGATCATATAGTTCCACAATCTTTCTTGAAAGATGATTCCATAGACAATAAAGTATTAACTAGAAGTGATAAAAATAGAGGTAAAAGTGATAATGTCCCAAGTGAGGAAGTCGTCAAAAAGATGAAAAATTACTGGCGTCAACTTTTGAATGCTAAATTAATTACTCAAAGAAAATTTGATAATTTGACTAAAGCAGAAAGAGGTGGGCTTTCTGAATTAGATAAAGCCGGGTTCATTAAAAGACAATTGGTCGAAACTAGACAAATTACTAAACATGTTGCCCAAATTTTAGATTCCCGTATGAACACTAAGTATGACGAAAATGATAAGTTAATACGTGAGGTTAAAGTCATTACTTTAAAATCAAAACTTGTCTCTGATTTCAGAAAGGATTTCCAATTCTATAAAGTTAGAGAAATTAATAATTATCATCATGCTCATGATGCATATTTGAATGCTGTAGTTGGAACTGCTTTAATCAAGAAATACCCTAAATTAGAATCTGAATTTGTATATGGTGATTACAAAGTCTATGATGTTAGAAAGATGATTGCTAAATCAGAACAAGAAATTGGTAAAGCTACAGCTAAATACTTCTTTTACTCTAACATTATGAATTTCTTTAAAACAGAAATTACTTTGGCAAACGGTGAAATTAGAAAAAGACCTCTTATTGAAACAAATGGTGAGACTGGAGAGATAGTTTGGGACAAAGGGCGTGATTTCGCTACTGTTAGAAAAGTTTTATCAATGCCACAAGTTAACATTGTAAAGAAAACAGAGGTTCAAACTGGTGGTTTCTCAAAAGAAAGTATTTTGCCTAAAAGAAATAGTGATAAATTGATTGCCAGAAAAAAGGATTGGGATCCAAAGAAATATGGTGGTTTCGACTCACCAACCGTAGCCTATTCTGTTTTGGTTGTGGCAAAGGTTGAAAAGGGTAAAAGTAAAAAGCTTAAATCAGTAAAAGAACTTTTGGGTATTACAATAATGGAAAGAAGTTCCTTTGAAAAGAACCCTATTGATTTTTTGGAAGCTAAAGGTTATAAGGAAGTAAAGAAGGACTTAATAATCAAATTGCCTAAATATTCTTTATTTGAATTAGAAAATGGGAGAAAAAGAATGTTGGCTTCTGCTGGAGAATTGCAAAAGGGTAATGAATTAGCATTGCCTTCCAAATATGTTAACTTCTTGTATTTAGCTTCACACTATGAAAAGTTGAAAGGGTCACCAGAAGATAACGAGCAAAAACAATTATTTGTTGAACAACACAAACACTACTTAGATGAGATTATAGAACAAATTAGTGAATTCAGTAAAAGAGTGATATTAGCTGATGCAAATTTAGATAAAGTTTTGTCAGCCTATAACAAACATAGAGATAAGCCAATTAGAGAACAAGCAGAAAACATTATTCACTTATTTACCCTTACCAATTTAGGAGCACCTGCTGCTTTCAAGTATTTTGATACAACAATTGATCGTAAAAGATATACCTCAACAAAAGAAGTCTTAGACGCCACCTTAATTCATCAATCAATCACTGGATTGTATGAGACAAGAATTGATTTGTCTCAATTGGGTGGTGAT
Protein Sequence
MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD
SV40 NLS nuclear localization signal of SV40 (si… 7680..7701 Fwd
DNA Sequence
CCTAAGAAGAAAAGAAAAGTT
Protein Sequence
PKKKRKV
SV40 NLS nuclear localization signal of SV40 (si… 7704..7725 Fwd
DNA Sequence
CCAAAGAAAAAGCGTAAGGTG
Protein Sequence
PKKKRKV
SV40 NLS nuclear localization signal of SV40 (si… 7728..7749 Fwd
DNA Sequence
CCTAAGAAAAAGAGAAAGGTT
Protein Sequence
PKKKRKV
3xFLAG three tandem FLAG(R) epitope tags, foll… 7749..7815 Fwd
DNA Sequence
GACTACAAAGACCATGACGGTGATTATAAAGATCATGACATCGACTACAAGGATGACGATGACAAG
Protein Sequence
DYKDHDGDYKDHDIDYKDDDDK
AmpR bla beta-lactamase 12943..13804 Rev
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
promoter 7
Label Gene Product Position Strand
URA3 promoter S. cerevisiae URA3 200..416 Fwd
DNA Sequence
TTCAATTCATCATTTTTTTTTTATTCTTTTTTTTGATTTCGGTTTCCTTGAAATTTTTTTGATTCGGTAATCTCCGAACAGAAGGAAGAACGAAGGAAGGAGCACAGACTTAGATTGGTATATATACGCATATGTAGTGTTGAAGAAACATGAAATTGCCCAGTATTCTTAACCCAACTGCACAGAACAAAAACCTGCAGGAAACGAAGATAAATC
T7 promoter 1972..1991 Fwd
DNA Sequence
TAATACGACTCACTATAGG
TEF promoter 8869..9213 Rev
DNA Sequence
GACATGGAGGCCCAGAATACCCTCCTTGACAGTCTTGACGTGCGCAGCTCAGGGGCATGATGTGACTGTCGCCCGTACATTTAGCCCATACATCCCCATGTATAATCATTTGCATCCATACATTTTGATGGCCGCACGGCGCGAAGCAAAAATTACGGCTCCTCGCTGCAGACCTGCGAGCAGGGAAACGCTCCCCTCACAGACGCGTTGAATTGTCCCCACGCCGCGCCCCTGTAGAGAAATATAAAAGGTTAGGATTTGCCACTGAGGTTCTTCTTTCATATACTTCCTTTTAAAATCTTGCTAGGATACAGTTCTCACATCACATCCGAACATAAACAACC
SNR52 promoter 9260..9529 Fwd
DNA Sequence
TCTTTGAAAAGATAATGTATGATTTTGCTTTCACTCATATTTATACAGAAACTTGATGTTTTCTTTCGAGTATATACAAGGTGATTACATGTACGTTTGAAGTACAACTCTAGATTTTGTAGTGCCCTCTTGGGCTAGCGGTAAAGGTGCGCATTTTTTCACACCCTACAATGTTCTGTTCAAAAGATTTTGGTCAAACGCTGTAGAAGTGAAAGTTGGTGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATC
T3 promoter 11743..11762 Rev
DNA Sequence
AATTAACCCTCACTAAAGG
lac promoter 11830..11861 Rev
DNA Sequence
TTTACACTTTATGCTTCCGGCTCGTATGTTG
AmpR promoter bla 13804..13909 Rev
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
protein_bind 2
Label Gene Product Position Strand
lac operator 11806..11823 Fwd
DNA Sequence
TTGTTATCCGCTCACAA
CAP binding site 11875..11897 Fwd
DNA Sequence
ATGAGTGAGCTAACTCACATTA
rep_origin 2
Label Gene Product Position Strand
f1 ori 1350..1806 Rev
DNA Sequence
ACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTT
ori 12184..12773 Rev
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 1
Label Gene Product Position Strand
TEF terminator 8071..8269 Rev
DNA Sequence
ACTGACAATAAAAAGATTCTTGTTTTCAAGAACTTGTCATTTGTATAGTTTTTTTATATTGTAGTTGTTCTATTTTAATCAAATGTTAGCGTGATTTATATTTTTTTTCGCCTCGACATCATCTGCCCAGATGCGAAGTTAAGTGCGCAGAAAGTAATATCATGCGTCAATCGTATGTGAATGCTGGTCGCTATACTG