pMS73

110629 Synthetic Bacterial Expression

Plasmid Information

Purpose
CRISPR/Cas9 vector for mutliplexed genome editing in Ustilago maydis. Expresses U. maydis codon-optimized Cas9 under the hsp70 promoter, contains a short version of U6 promoter, is self-replicating
Depositor
Regine Kahmann
Article
Schuster et al Fungal Genet Biol. 2018 Mar;112:21-30. d
Tags
N-terminal NLS, C-terminal HA-tag +NLS
Mutation
Streptococcus pyogenes gene codon-optimized for U…
Promoter
U. maydis hsp70 promoter (Kronstad and Leong, 198…
Available Since
May 16, 2018
Availability
Available

Sequence Information

Length
10,783 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 5
Label Gene Product Position Strand
SV40 NLS — nuclear localization signal of SV40 (si… 1712..1733 Fwd
DNA Sequence
CCTAAGAAGAAACGCAAGGTT
Protein Sequence
PKKKRKV
Cas9 — Cas9 (Csn1) endonuclease from the Strep… 1736..5837 Fwd
DNA Sequence
GATAAGAAGTACAGCATCGGACTCGACATCGGTACTAACTCGGTAGGATGGGCAGTCATCACGGATGAATACAAGGTTCCTTCCAAGAAGTTTAAGGTCCTTGGTAACACCGACCGCCATTCTATCAAGAAGAACCTCATTGGCGCTTTGCTCTTTGACTCAGGAGAAACCGCTGAGGCGACACGCCTCAAACGCACGGCACGTCGACGTTATACACGCAGAAAGAATCGTATCTGCTATCTGCAGGAAATCTTTTCGAACGAAATGGCAAAAGTTGATGACAGCTTCTTCCATCGCCTGGAGGAATCGTTTCTCGTGGAGGAGGACAAGAAGCACGAGAGACATCCTATCTTCGGCAACATTGTCGATGAGGTCGCTTACCACGAGAAGTACCCTACTATCTACCACCTTAGAAAGAAGCTCGTAGACTCAACTGACAAAGCGGATCTTCGTCTGATCTATTTGGCTCTTGCCCACATGATCAAGTTCCGTGGTCATTTTCTCATCGAAGGCGACCTTAATCCCGACAACTCGGACGTCGATAAGCTGTTTATCCAGCTCGTACAGACATACAACCAGCTCTTCGAAGAAAACCCAATTAATGCTTCCGGTGTTGATGCAAAAGCTATCCTCAGCGCGAGATTGAGCAAGAGCAGACGCCTCGAAAACCTCATCGCCCAATTGCCTGGTGAAAAGAAGAACGGCTTGTTCGGCAATCTTATTGCCCTGAGCCTTGGCCTCACTCCGAACTTCAAGTCGAACTTCGATCTTGCGGAAGATGCCAAGCTGCAACTCTCCAAGGACACGTACGATGATGACTTGGACAATCTCCTTGCCCAGATTGGCGATCAATACGCCGATCTTTTCCTCGCGGCGAAGAACTTGTCGGACGCAATCTTGCTCTCAGACATCCTTCGCGTCAACACTGAGATCACCAAAGCCCCTCTCTCTGCCTCGATGATCAAGCGCTATGACGAACACCACCAGGATCTCACGCTCCTTAAGGCATTGGTGCGTCAGCAGTTGCCTGAGAAGTACAAAGAGATTTTCTTTGATCAGTCGAAGAACGGATACGCTGGCTACATCGACGGTGGCGCTTCTCAGGAGGAGTTCTACAAGTTTATCAAACCCATTCTTGAGAAGATGGATGGCACGGAGGAGCTCCTCGTCAAGCTGAATCGCGAGGACCTCCTCCGTAAGCAACGTACGTTCGACAATGGCTCGATTCCACACCAGATTCATCTGGGCGAACTCCACGCCATCCTCAGGAGGCAGGAGGACTTCTATCCCTTCCTCAAGGATAATCGAGAGAAAATTGAGAAGATCCTCACATTCCGCATCCCCTATTATGTAGGCCCACTCGCTCGCGGAAACTCTCGCTTTGCCTGGATGACCCGCAAGTCGGAAGAAACAATCACCCCGTGGAACTTCGAAGAGGTGGTGGACAAGGGTGCATCTGCGCAGTCGTTTATTGAGAGGATGACAAACTTTGATAAGAACCTCCCGAATGAGAAAGTCCTGCCAAAACATTCCCTCCTGTATGAATACTTCACGGTCTATAACGAACTGACAAAGGTGAAGTACGTGACCGAGGGTATGCGTAAGCCTGCCTTTCTTTCGGGTGAGCAGAAGAAAGCTATTGTCGACTTGTTGTTCAAGACCAACCGCAAGGTCACTGTCAAGCAACTGAAGGAAGATTACTTCAAGAAAATCGAGTGTTTTGATTCGGTAGAGATCTCGGGCGTCGAGGACAGGTTCAACGCCTCTCTCGGCACCTATCACGATCTTCTCAAGATCATCAAGGACAAAGACTTTCTTGACAACGAAGAGAACGAGGATATTCTCGAGGACATCGTGCTCACCCTCACTTTGTTCGAAGATCGCGAAATGATTGAGGAACGTCTTAAGACATATGCTCACTTGTTCGACGACAAAGTGATGAAGCAGCTGAAGCGTAGGCGATACACAGGTTGGGGCCGCCTCTCGCGCAAGCTGATTAACGGTATCCGCGACAAGCAATCCGGCAAGACAATCTTGGATTTCCTTAAGAGCGACGGTTTTGCTAACCGCAACTTCATGCAGCTCATCCACGACGACAGCCTTACGTTCAAGGAGGACATCCAGAAGGCCCAGGTTTCCGGACAAGGTGACTCTCTCCATGAGCACATCGCTAACCTGGCGGGAAGCCCCGCGATCAAGAAAGGTATCCTCCAGACCGTCAAAGTTGTGGACGAGCTGGTCAAGGTAATGGGCCGACACAAACCGGAGAACATTGTTATCGAGATGGCACGAGAGAATCAGACGACCCAGAAAGGCCAAAAGAACTCCAGAGAACGTATGAAACGAATCGAAGAGGGTATCAAGGAACTGGGATCGCAGATCCTGAAGGAGCACCCCGTTGAGAACACGCAGCTCCAAAACGAAAAGCTGTACCTCTACTACTTGCAAAATGGTAGGGATATGTACGTCGACCAGGAACTGGATATTAATCGTCTGTCCGACTACGACGTTGACCACATCGTGCCCCAATCGTTTCTCAAGGATGACTCGATCGATAATAAAGTACTTACGCGCTCAGACAAGAACCGAGGTAAATCGGACAATGTCCCATCGGAGGAAGTCGTGAAGAAGATGAAGAACTATTGGCGCCAACTTCTTAACGCAAAGCTGATCACCCAGAGGAAATTCGACAACCTCACCAAAGCAGAACGCGGCGGCCTCTCCGAGCTCGACAAGGCTGGATTTATCAAGCGTCAGCTCGTCGAAACGCGTCAGATTACCAAGCACGTCGCACAGATCCTGGATAGCCGCATGAACACAAAGTACGACGAAAACGACAAGCTCATCCGTGAGGTTAAGGTCATCACCTTGAAGTCGAAACTCGTGTCGGACTTCCGCAAAGATTTTCAGTTCTATAAAGTTAGAGAGATCAACAACTACCACCATGCGCATGACGCCTACCTCAATGCCGTCGTGGGCACCGCACTTATTAAGAAATACCCGAAGCTCGAGTCCGAGTTTGTCTACGGCGATTACAAGGTATACGACGTTCGCAAGATGATTGCCAAATCGGAGCAGGAGATCGGTAAGGCCACTGCCAAGTACTTCTTTTACTCGAACATCATGAATTTCTTCAAAACAGAAATCACCCTCGCCAACGGCGAGATTCGCAAACGACCACTCATCGAGACTAACGGTGAAACGGGAGAGATCGTCTGGGATAAGGGCCGAGACTTTGCTACGGTTCGAAAGGTCCTTTCGATGCCTCAAGTGAACATCGTCAAGAAAACGGAGGTCCAAACCGGTGGCTTCAGCAAGGAGTCGATTCTGCCGAAACGCAATTCGGACAAATTGATTGCACGCAAGAAGGATTGGGACCCTAAGAAATATGGCGGCTTCGATTCACCGACAGTGGCCTATTCGGTTCTGGTCGTCGCGAAAGTGGAGAAGGGCAAGTCAAAGAAGCTCAAGTCAGTGAAGGAGCTCCTGGGAATCACCATCATGGAACGTTCCTCTTTTGAGAAGAACCCTATCGACTTTCTCGAGGCTAAGGGCTACAAAGAGGTCAAGAAAGATCTCATCATCAAACTCCCAAAATACTCACTTTTCGAGCTCGAGAACGGCCGTAAACGAATGCTGGCGAGCGCAGGAGAGCTTCAAAAGGGAAATGAACTGGCTTTGCCCTCCAAGTACGTCAACTTCCTCTACCTCGCAAGCCATTATGAGAAGTTGAAGGGTAGCCCCGAGGACAACGAACAAAAGCAGCTCTTCGTGGAGCAACACAAACATTACCTGGATGAAATCATCGAGCAAATCTCGGAGTTTAGCAAGCGAGTGATCTTGGCTGATGCCAACCTCGACAAGGTGTTGTCTGCCTACAACAAGCATCGAGATAAGCCGATTCGCGAGCAGGCCGAGAACATCATCCACCTCTTCACTCTCACTAACTTGGGTGCGCCTGCGGCGTTTAAATACTTTGACACGACCATCGACCGCAAGCGTTACACAAGCACGAAGGAAGTCCTCGACGCTACACTGATCCATCAGTCGATCACCGGTCTGTACGAAACCCGCATCGATCTGTCTCAACTGGGCGGTGAC
Protein Sequence
DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD
HA — HA (human influenza hemagglutinin) epit… 5846..5873 Fwd
DNA Sequence
TACCCATACGATGTGCCCGATTACGCT
Protein Sequence
YPYDVPDYA
nucleoplasmin NLS — bipartite nuclear localization signal f… 5882..5930 Fwd
DNA Sequence
AAGCGTCCCGCAGCCACTAAGAAGGCTGGACAGGCCAAGAAGAAGAAG
Protein Sequence
KRPAATKKAGQAKKKK
AmpR bla beta-lactamase 9066..9927 Fwd
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
promoter 1
Label Gene Product Position Strand
lac promoter — — 226..257 Fwd
DNA Sequence
TTTACACTTTATGCTTCCGGCTCGTATGTTG
protein_bind 2
Label Gene Product Position Strand
CAP binding site — — 190..212 Fwd
DNA Sequence
TAATGTGAGTTAGCTCACTCAT
lac operator — — 264..281 Fwd
DNA Sequence
TTGTGAGCGGATAACAA
rep_origin 1
Label Gene Product Position Strand
ori — — 10097..10686 Fwd
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 1
Label Gene Product Position Strand
NOS terminator — — 5950..6203 Fwd
DNA Sequence
GATCGTTCAAACATTTGGCAATAAAGTTTCTTAAGATTGAATCCTGTTGCCGGTCTTGCGATGATTATCATATAATTTCTGTTGAATTACGTTAAGCATGTAATAATTAACATGTAATGCATGACGTTATTTATGAGATGGGTTTTTATGATTAGAGTCCCGCAATTATACATTTAATACGCGATAGAAAACAAAATATAGCGCGCAAACTAGGATAAATTATCGCGCGCGGTGTCATCTATGTTACTAGATC