pDual

207883 Saccharomyces cerevisiae Bacterial Expression

Plasmid Information

Purpose
Expresses thermosensitive variants of the homing endonucleases I-OnuI and I-GpeMI, each in a different open reading frame. Both were made thermosensitive by a fusion with the L212P VMA1 intein.
Depositor
David Edgell
Article
Foo et al Nucleic Acids Res. 2024 Feb 28;52(4):206
Mutation
Leucine 212 changed to proline, leucine 434 in th…
Available Since
Aug. 29, 2024
Availability
Available

Sequence Information

Length
7,785 bp
Topology
circular
Molecule Type
other DNA

Features

CDS 3
Label Gene Product Position Strand
Sce VMA intein 5' region Saccharomyces cerevisiae VMA1 5' region of the intein from the yeast … 1669..2488 Fwd
DNA Sequence
TGCTTTGCCAAGGGTACCAATGTTTTAATGGCGGATGGGTCTATTGAATGTATTGAAAACATTGAGGTTGGTAATAAGGTCATGGGTAAAGATGGCAGACCTCGTGAGGTAATTAAATTGCCCAGAGGAAGAGAAACTATGTACAGCGTCGTGCAGAAAAGTCAGCACAGAGCCCACAAAAGTGACTCAAGTCGTGAAGTGCCAGAATTACTCAAGTTTACGTGTAATGCGACCCATGAGTTGGTTGTTAGAACACCTCGTAGTGTCCGCCGTTTGTCTCGTACCATTAAGGGTGTCGAATATTTTGAAGTTATTACTTTTGAGATGGGCCAAAAGAAAGCCCCCGACGGTAGAATTGTTGAGCTTGTCAAGGAAGTTTCAAAGAGCTACCCAATATCTGAGGGGCCTGAGAGAGCCAACGAATTAGTAGAATCCTATAGAAAGGCTTCAAATAAAGCTTATTTTGAGTGGACTATTGAGGCCAGAGATCTTTCTCTGTTGGGTTCCCATGTTCGTAAAGCTACCTACCAGACTTACGCTCCAATTCTTTATGAGAATGACCACTTTTTCGACTACATGCAAAAAAGTAAGTTTCATCTCACCATTGAAGGTCCAAAAGTACTTGCTTATTTACCTGGTTTATGGATTGGTGATGGATTGTCTGACAGGGCAACTTTTTCGGTTGATTCCAGAGATACTTCTTTGATGGAACGTGTTACTGAATATGCTGAAAAGTTGAATTTGTGCGCCGAGTATAAGGACAGAAAAGAACCACAAGTTGCCAAAACTGTTAATTTGTACTCTAAAGTTGTCAGAGGT
Protein Sequence
CFAKGTNVLMADGSIECIENIEVGNKVMGKDGRPREVIKLPRGRETMYSVVQKSQHRAHKSDSSREVPELLKFTCNATHELVVRTPRSVRRLSRTIKGVEYFEVITFEMGQKKAPDGRIVELVKEVSKSYPISEGPERANELVESYRKASNKAYFEWTIEARDLSLLGSHVRKATYQTYAPILYENDHFFDYMQKSKFHLTIEGPKVLAYLPGLWIGDGLSDRATFSVDSRDTSLMERVTEYAEKLNLCAEYKDRKEPQVAKTVNLYSKVVRG
Factor Xa site Factor Xa recognition and cleavage site 2943..2955 Fwd
DNA Sequence
ATTGAAGGAAGA
Protein Sequence
IEGR
AmpR bla beta-lactamase 6595..7456 Fwd
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGCAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
promoter 2
Label Gene Product Position Strand
BBa_J23100 3622..3657 Fwd
DNA Sequence
TTGACGGCTAGCTCAGTCCTAGGTACAGTGCTAGC
AmpR promoter bla 6490..6595 Fwd
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
rep_origin 1
Label Gene Product Position Strand
p15A ori 0..7785 Fwd
DNA Sequence
TTGAGATCGTTTTGGTCTGCGCGTAATCTCTTGCTCTGAAAACGAAAAAACCGCCTTGCAGGGCGGTTTTTCGAAGGTTCTCTGAGCTACCAACTCTTTGAACCGAGGTAACTGGCTTGGAGGAGCGCAGTCACCAAAACTTGTCCTTTCAGTTTAGCCTTAACCGGCGCATGACTTCAAGACTAACTCCTCTAAATCAATTACCAGTGGCTGCTGCCAGTGGTGCTTTTGCATGTCTTTCCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGACTGAACGGGGGGTTCGTGCATACAGTCCAGCTTGGAGCGAACTGCCTACCCGGAACTGAGTGTCAGGCGTGGAATGAGACAAACGCGGCCATAACAGCGGAATGACACCGGTAAACCGAAAGGCAGGAACAGGAGAGCGCACGAGGGAGCCGCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCACTGATTTGAGCGTCAGATTTCGTGATGCTTGTCAGGGGGGCGGAGCCTATGGAAA