pET23PI-Sce ESARC D218A

195608 Saccharomyces cerevisiae Bacterial Expression

Plasmid Information

Purpose
Expresses PI-SceI (VDE) homing endonuclease/intein from the T7 promoter that contains the D218A active site mutation
Depositor
Frederick Gimble
Article
Gimble et al J Biol Chem. 1995 Mar 17;270(11):5849-56
Insert
PI-SceI
Mutation
T928C, T929C, A942C
Promoter
T7
Available Since
Feb. 1, 2023
Availability
Available

Sequence Information

Length
5,027 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 6
Label Gene Product Position Strand
Sce VMA intein 5' region Saccharomyces cerevisiae VMA1 5' region of the intein from the yeast … 295..1114 Fwd
DNA Sequence
TGCTTTGCCAAGGGTACCAATGTTTTAATGGCGGATGGGTCTATTGAATGTATTGAAAACATTGAGGTTGGTAATAAGGTCATGGGTAAAGATGGCAGACCTCGTGAGGTAATTAAATTGCCCAGAGGAAGAGAAACTATGTACAGCGTCGTGCAGAAAAGTCAGCACAGAGCCCACAAAAGTGACTCAAGTCGTGAAGTGCCAGAATTACTCAAGTTTACGTGTAATGCGACCCATGAGTTGGTTGTTAGAACACCTCGTAGTGTCCGCCGTTTGTCTCGTACCATTAAGGGTGTCGAATATTTTGAAGTTATTACTTTTGAGATGGGCCAAAAGAAAGCCCCCGACGGTAGAATTGTTGAGCTTGTCAAGGAAGTTTCAAAGAGCTACCCAATATCTGAGGGGCCTGAGAGAGCCAACGAATTAGTAGAATCCTATAGAAAGGCTTCAAATAAAGCTTATTTTGAGTGGACTATTGAGGCCAGAGATCTTTCTCTGTTGGGTTCCCATGTTCGTAAAGCTACCTACCAGACTTACGCTCCAATTCTTTATGAGAATGACCACTTTTTCGACTACATGCAAAAAAGTAAGTTTCATCTCACCATTGAAGGTCCAAAAGTACTTGCTTATCTCCTAGGCCTATGGATTGGTGCTGGATTGTCGGACCGGGCAACTTTTTCGGTTGATTCCAGAGATACTTCTTTGATGGAACGTGTTACTGAATATGCTGAAAAGTTGAATTTGTGCGCCGAGTATAAGGACAGAAAAGAACCACAAGTTGCCAAAACTGTTAATTTGTACTCTAAAGTTGTCAGAGGT
Protein Sequence
CFAKGTNVLMADGSIECIENIEVGNKVMGKDGRPREVIKLPRGRETMYSVVQKSQHRAHKSDSSREVPELLKFTCNATHELVVRTPRSVRRLSRTIKGVEYFEVITFEMGQKKAPDGRIVELVKEVSKSYPISEGPERANELVESYRKASNKAYFEWTIEARDLSLLGSHVRKATYQTYAPILYENDHFFDYMQKSKFHLTIEGPKVLAYLLGLWIGAGLSDRATFSVDSRDTSLMERVTEYAEKLNLCAEYKDRKEPQVAKTVNLYSKVVRG
Factor Xa site — Factor Xa recognition and cleavage site 1569..1581 Fwd
DNA Sequence
ATTGAAGGAAGA
Protein Sequence
IEGR
T7 tag (gene 10 leader) — leader peptide from bacteriophage T7 ge… 1660..1690 Fwd
DNA Sequence
GCTAGCATGACTGGTGGACAGCAAATGGGT
Protein Sequence
ASMTGGQQMG
6xHis — 6xHis affinity tag 1739..1757 Fwd
DNA Sequence
CACCACCACCACCACCAC
Protein Sequence
HHHHHH
AmpR bla beta-lactamase 2493..3354 Fwd
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGCAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
rop rop Rop protein, which maintains plasmids a… 4539..4731 Rev
DNA Sequence
GTGACCAAACAGGAAAAAACCGCCCTTAACATGGCCCGCTTTATCAGAAGCCAGACATTAACGCTTCTGGAGAAACTCAACGAGCTGGACGCGGATGAACAGGCAGACATCTGTGAATCGCTTCACGACCACGCTGATGAGCTTTACCGCAGCTGCCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGA
Protein Sequence
MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHADELYRSCLARFGDDGENL
promoter 2
Label Gene Product Position Strand
T7 promoter — — 212..231 Fwd
DNA Sequence
TAATACGACTCACTATAGG
AmpR promoter bla — 2388..2493 Fwd
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
rep_origin 2
Label Gene Product Position Strand
f1 ori — — 1907..2363 Fwd
DNA Sequence
ACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTT
ori — — 3524..4113 Fwd
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 1
Label Gene Product Position Strand
T7 terminator — — 1823..1871 Fwd
DNA Sequence
CTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG