pIW447-CRISPR XYL2

98906 Synthetic Yeast Expression

Plasmid Information

Purpose
K. marxianus CRISPR Plasmid for XYL2 editing
Depositor
Ian Wheeldon
Article
Lobs et al Biotechnol Biofuels. 2017 Jun 24;10:164.
Tags
SV40
Available Since
Oct. 25, 2017
Availability
Available

Sequence Information

Length
11,129 bp
Topology
circular
Molecule Type
other DNA

Features

CDS 4
Label Gene Product Position Strand
Cas9(C) — C-terminal portion of Streptococcus pyo… 2340..4725 Fwd
DNA Sequence
TGTTTCGACTCTGTCGAAATTTCTGGTGTCGAAGACAGATTCAATGCTTCTTTGGGTACTTACCACGATTTGTTGAAAATCATTAAGGATAAGGATTTCTTAGATAACGAGGAGAACGAAGATATTTTGGAGGATATTGTCTTGACTTTGACTTTGTTTGAGGATAGAGAAATGATCGAGGAAAGATTGAAGACTTACGCTCATTTGTTCGATGATAAGGTCATGAAGCAATTGAAGAGAAGAAGATACACTGGTTGGGGTAGATTGTCCAGAAAGTTGATCAACGGTATTAGAGATAAGCAATCCGGTAAGACCATTTTGGATTTCTTGAAGTCTGACGGTTTCGCTAATAGAAACTTTATGCAATTGATTCACGACGATTCTTTGACTTTTAAGGAAGATATTCAAAAGGCTCAAGTCTCTGGTCAGGGTGATTCTTTGCACGAACACATCGCTAACTTGGCTGGTTCTCCAGCTATCAAAAAAGGTATTTTGCAAACTGTTAAGGTTGTTGACGAATTGGTTAAGGTTATGGGTAGACACAAGCCAGAGAATATTGTCATCGAGATGGCTAGAGAAAATCAAACCACTCAAAAGGGTCAAAAGAACTCCAGAGAGAGAATGAAAAGAATTGAGGAAGGTATCAAGGAGTTGGGTTCCCAAATTTTGAAGGAACACCCAGTCGAGAACACTCAATTGCAAAATGAAAAGTTGTACTTGTATTACTTGCAAAACGGTAGAGATATGTACGTTGACCAAGAATTGGACATTAACAGATTGTCTGACTACGATGTTGATCACATCGTTCCACAATCCTTTTTGAAGGATGACTCTATCGATAACAAGGTTTTGACTAGATCCGACAAGAATAGAGGTAAGTCTGATAACGTTCCATCTGAAGAAGTCGTCAAGAAGATGAAGAACTACTGGAGACAGTTGTTGAATGCTAAGTTGATTACTCAAAGAAAGTTCGATAACTTGACTAAGGCTGAGAGAGGTGGTTTGTCTGAGTTGGATAAGGCCGGTTTCATTAAGAGACAATTGGTCGAAACCAGACAAATCACCAAGCACGTTGCTCAAATCTTGGACTCTAGAATGAACACCAAGTACGACGAAAACGACAAATTGATTAGAGAGGTTAAGGTCATCACCTTGAAGTCCAAATTGGTTTCCGACTTTAGAAAGGATTTCCAATTCTACAAGGTCAGAGAAATCAATAACTACCACCATGCTCACGACGCCTACTTGAACGCTGTTGTCGGTACTGCTTTGATCAAGAAGTACCCAAAGTTGGAGTCTGAATTTGTCTACGGTGATTACAAAGTTTACGACGTTAGAAAGATGATTGCTAAGTCTGAGCAAGAAATCGGTAAGGCTACTGCTAAATACTTCTTCTACTCCAACATTATGAATTTCTTCAAGACCGAGATTACCTTGGCCAACGGTGAAATCAGAAAGAGACCATTGATTGAAACTAACGGTGAAACCGGTGAGATTGTCTGGGACAAGGGTAGAGATTTCGCTACTGTTAGAAAGGTCTTGTCTATGCCACAAGTCAACATTGTCAAGAAGACCGAAGTCCAGACTGGTGGTTTCTCCAAGGAATCTATTTTGCCAAAGAGAAACTCTGATAAGTTGATTGCCAGAAAGAAAGACTGGGATCCAAAGAAGTACGGTGGTTTCGACTCTCCAACCGTTGCTTACTCCGTTTTGGTTGTCGCTAAGGTTGAAAAGGGTAAGTCCAAGAAGTTGAAGTCCGTCAAGGAGTTGTTGGGTATTACCATCATGGAAAGATCTTCCTTCGAAAAGAACCCAATTGATTTCTTGGAGGCTAAGGGTTACAAGGAAGTCAAGAAGGACTTGATCATTAAGTTGCCAAAGTACTCTTTGTTCGAATTGGAGAACGGTAGAAAGAGAATGTTGGCTTCCGCTGGTGAATTGCAAAAGGGTAACGAATTGGCTTTGCCATCTAAGTACGTTAACTTCTTGTACTTGGCTTCCCACTACGAAAAGTTGAAGGGTTCTCCAGAAGATAACGAACAAAAGCAATTGTTCGTTGAACAACATAAGCACTACTTGGACGAAATCATCGAACAAATCTCTGAATTCTCTAAGAGAGTTATCTTGGCTGACGCTAACTTGGACAAGGTTTTGTCTGCTTACAACAAGCACAGAGACAAGCCAATCAGAGAGCAAGCTGAAAACATCATCCACTTGTTCACTTTGACCAACTTGGGTGCTCCAGCTGCTTTCAAGTACTTCGACACCACCATCGACAGAAAGAGATACACCTCTACCAAGGAAGTTTTGGACGCTACCTTGATCCACCAATCTATCACCGGTTTGTACGAAACCAGAATCGACTTGTCTCAATTGGGTGGTGAC
Protein Sequence
CFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD
SV40 NLS — nuclear localization signal of SV40 (si… 4737..4758 Fwd
DNA Sequence
CCAAAGAAGAAGAGAAAGGTT
Protein Sequence
PKKKRKV
AmpR bla beta-lactamase 6970..7831 Rev
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
URA3 S. cerevisiae URA3 orotidine-5'-phosphate decarboxylase, r… 9732..10536 Fwd
DNA Sequence
ATGTCGAAAGCTACATATAAGGAACGTGCTGCTACTCATCCTAGTCCTGTTGCTGCCAAGCTATTTAATATCATGCACGAAAAGCAAACAAACTTGTGTGCTTCATTGGATGTTCGTACCACCAAGGAATTACTGGAGTTAGTTGAAGCATTAGGTCCCAAAATTTGTTTACTAAAAACACATGTGGATATCTTGACTGATTTTTCCATGGAGGGCACAGTTAAGCCGCTAAAGGCATTATCCGCCAAGTACAATTTTTTACTCTTCGAAGACAGAAAATTTGCTGACATTGGTAATACAGTCAAATTGCAGTACTCTGCGGGTGTATACAGAATAGCAGAATGGGCAGACATTACGAATGCACACGGTGTGGTGGGCCCAGGTATTGTTAGCGGTTTGAAGCAGGCGGCAGAAGAAGTAACAAAGGAACCTAGAGGCCTTTTGATGTTAGCAGAATTGTCATGCAAGGGCTCCCTATCTACTGGAGAATATACTAAGGGTACTGTTGACATTGCGAAGAGCGACAAAGATTTTGTTATCGGCTTTATTGCTCAAAGAGACATGGGTGGAAGAGATGAAGGTTACGATTGGTTGATTATGACACCCGGTGTGGGTTTAGATGACAAGGGAGACGCATTGGGTCAACAGTATAGAACCGTGGATGATGTGGTCTCTACAGGATCTGACATTATTATTGTTGGAAGAGGACTATTTGCAAAGGGAAGGGATGCTAAGGTAGAGGGTGAACGTTACAGAAAAGCAGGCTGGGAAGCATATTTGAGAAGATGCGGCCAGCAAAACTAA
Protein Sequence
MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELLELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQYSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGEYTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDDVVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN
promoter 6
Label Gene Product Position Strand
T7 promoter — — 156..175 Fwd
DNA Sequence
TAATACGACTCACTATAGG
TEF1 promoter S. cerevisiae TEF1 — 191..590 Fwd
DNA Sequence
CCTAGCTTCAAAATGTTTCTACTCCTTTTTTACTCTTCCAGATTTTCTCGGACTCCGCGCATCGCCGTACCACTTCAAAACACCCAAGCACAGCATACTAAATTTCCCCTCTTTCTTCCTCTAGGGTGTCGTTAATTACCCGTACTAAAGGTTTGGAAAAGAAAAAAGAGACCGCCTCGTTTCTTTTTCTTCGTCGAAAAAGGCAATAAAAATTTTTATCACGTTTCTTTTTCTTGAAAATTTTTTTTTTGATTTTTTTCTCTTTCGATGACCTCCCATTGATATTTAAGTTAATAAACGGTCTTCAATTTCTCAAGTTTCAGTTTCATTTTTCTTGTTCTATTACAACTTTTTTTACTTCTTGCTCATTAGAAAGAAAGCATAGCAATCTAATCTAAG
T3 promoter — — 5770..5789 Rev
DNA Sequence
AATTAACCCTCACTAAAGG
lac promoter — — 5857..5888 Rev
DNA Sequence
TTTACACTTTATGCTTCCGGCTCGTATGTTG
AmpR promoter bla — 7831..7936 Rev
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
URA3 promoter S. cerevisiae URA3 — 9516..9732 Fwd
DNA Sequence
TTCAATTCATCATTTTTTTTTTATTCTTTTTTTTGATTTCGGTTTCCTTGAAATTTTTTTGATTCGGTAATCTCCGAACAGAAGGAAGAACGAAGGAAGGAGCACAGACTTAGATTGGTATATATACGCATATGTAGTGTTGAAGAAACATGAAATTGCCCAGTATTCTTAACCCAACTGCACAGAACAAAAACCTGCAGGAAACGAAGATAAATC
protein_bind 2
Label Gene Product Position Strand
lac operator — — 5833..5850 Fwd
DNA Sequence
TTGTTATCCGCTCACAA
CAP binding site — — 5902..5924 Fwd
DNA Sequence
ATGAGTGAGCTAACTCACATTA
rep_origin 2
Label Gene Product Position Strand
ori — — 6211..6800 Rev
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
f1 ori — — 10666..11122 Rev
DNA Sequence
ACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTT
terminator 2
Label Gene Product Position Strand
CYC1 terminator S. cerevisiae CYC1 — 4773..5021 Fwd
DNA Sequence
TCATGTAATTAGTTATGTCACGCTTACATTCACGCCCTCCCCCCACATCCGCTCTAACCGAAAAGGAAGGAGTTAGACAACCTGAAGTCTAGGTCCCTATTTATTTTTTTATAGTTATGTTAGTATTAAGAACGTTATTTATATTTCAAATTTTTCTTTTTTTTCTGTACAGACGCGTGTACGCATGTAACATTATACTGAAAACCTTGCTTGAGAAGGTTTTGGGACGCTCGAAGGCTTTAATTTGC
SUP4 terminator — — 5726..5746 Fwd
DNA Sequence
TTTTTTTGTTTTTTATGTCT