Gene:
Gene Symbol:
Gene Name:
Original Clone ID:
2482.1.08_GO.74368
Keyword:
None
Species:
Saccharomyces cerevisiae
Type:
cDNA
Vector Name:
pVP65K
Format:
CLOSED
Source:
Center for Eukaryotic Structural Genomics
Description:
None
Comments:
MBP-His8 fusion protein; MPB is cleaved in vivo by TVMV protease. TEV cleavage site that is downstream from the SgfI site is used to remove His8 tag. after TEV cleavage the protein obtained will be the translated CDS with the addition of S
Mutation / Discrepancy:
No / No
Publications:
PMID: 17320109
Title: Structure of pyrimidine 5'-nucleotidase type 1. Insight into mechanism of action and inhibition during lead poisoning.
Title: Structure of pyrimidine 5'-nucleotidase type 1. Insight into mechanism of action and inhibition during lead poisoning.
Authors:
University of Wisconsin Madison
PSI
Center for Eukaryotic Structural Genomics
PSI
Center for Eukaryotic Structural Genomics
ScCD00297618
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AVAILABLE
No restriction
Special MTA: PSI
Special MTA: PSI
Insert sequence: 681nts
Open reading frame: 50 to 730
GTACTTCCAGTCCGAGTATGGACATCACGCTAGACCAGATTCAAAACGACCATTGGATGA
AGGATCACCAGCAGCCGCAGGATTAACTTCTAAGAAGGCGAACGAAGCTTTAACAAGAAA
TAGAGAATTAACAACTGTATTAGTCAAAAATTTGCCTAAGAGCTATAATCAAAACAAAGT
CTATAAGTACTTCAAACATTGTGGGCCCATTATACACGTTGATGTTGCGGATTCGCTGAA
GAAGAACTTTCGTTTTGCACGTATTGAATTTGCCAGGTATGATGGAGCCCTCGCTGCAAT
AACCAAAACACACAAAGTTGTAGGTCAGAATGAAATTATAGTATCTCATTTAACAGAATG
CACATTATGGATGACGAATTTCCCCCCAAGTTATACTCAAAGAAATATTAGAGATCTATT
GCAAGATATTAACGTTGTTGCACTCAGTATACGGCTTCCCAGTTTACGATTCAATACAAG
CAGAAGGTTCGCTTACATCGATGTTACTTCTAAAGAGGATGCAAGATATTGTGTAGAAAA
ATTGAATGGACTCAAAATAGAAGGTTATACTTTAGTTACTAAAGTTTCCAATCCGCTGGA
AAAGTCGAAACGGACCGATTCTGCCACACTAGAGGGGCGAGAGATTATGATACGAAACCT
AAGTACAGAATTGCTCGACGAAAACCTTCTGAGGGAATCTTTTGAGGGATTTGGTTCCAT
TGAAAAATAGTTTAAACGAATTCGAGCTCGGTACCCGGGGATCCTCTAGAGTCGACCTG
Protein sequence predicted based on nucleotide sequence
PLDEGSPAAAGLTSKKANEALTRNRELTTVLVKNLPKSYNQNKVYKYFKHCGPIIHVDVA DSLKKNFRFARIEFARYDGALAAITKTHKVVGQNEIIVSHLTECTLWMTNFPPSYTQRNI RDLLQDINVVALSIRLPSLRFNTSRRFAYIDVTSKEDARYCVEKLNGLKIEGYTLVTKVS NPLEKSKRTDSATLEGREIMIRNLSTELLDENLLRESFEGFGSIEK*
Coding Sequence Details
Insert Sequence Verified?:
Y
Verification Method:
Sequence Verification
Reference Sequence Annotations:
Start on reference sequence
1
End on reference sequence
750
Mutations:
None
Discrepancy:
None
5' Linker Sequence:
None
3' Linker Sequence:
None
Recommended Growth Condition:
Distributed in bacterial strain:
DH5-alpha T1 phage resistant
Antibiotic Selection:
Host Type:
bacterial
Marker:
kanamycin
Bacterial Selection Condition:
50ug/mL kanamycin
Growth Condition:
Growth with the single antibiotic in LB at 37 degrees is recommended.
Comments:
Conditions for Gateway-type vectors in recombined (with insert) form and other kanamycin resistant vectors.
Protein Expression Results:
ProteinExpressed: Protein_Confirmed
ProteinPurified: Protein_Purified
SolubleProtein: Protein_Soluble
Find experimental protocols in GO.74368
Recommended expression in:
Not Applicable
Vector Name:
pVP65K
Synonyms:
''
Sequencing Primer:
Forward:
MBP Forward
Reverse:
Unknown
Description:
Bacterial (E. coli) expression with N-terminal 8xHis, MBP tag; kanamycin resistance in bacteria; restriction enzyme cloning (SgfI, PmeI); Cloning into this vector will remove the BarCat cassette.
Comments:
Vector contains a lethal BarCat cassette that requires the strain BR610 for growth. Lethal cassette is removed during cloning.
Size (bp):
7174
Parent Vector:
None
Properties:
bacterial expression
Author Name:
University of Wisconsin
Center for Eukaryotic Structural Genomics
PSI
Center for Eukaryotic Structural Genomics
PSI
Publications:
PMID:
18988018
Title: Flexi vector cloning.
Vector Map:
Vector Sequence:
Sequence:
Vector Features:
Type
Name
Description
Start
End
bacterial origin
pBR322 origin
pBR322 bacterial origin of replication
4977
6204
gene fragment
TVMV cds
TVMV gene
139
849
gene fragment
BarCat
BarCat cassette including Barnase
2245
3376
primer
MBP forward
MBP forward primer
2071
2091
primer
T7 term
T7 terminator reverse primer
3426
3443
promoter
T5
T5 promoter region
952
1032
promoter
TET
TET promoter
873
946
promoter
LacIq
LacIq-wt promoter
4899
4967
protease cleavage site
TVMV
TVMV protease cleavage site
2189
2209
repressor binding site
LacI
LacI coding region
3807
4898
selectable marker
KanR
Kanamycin resistance gene
6213
7174
tag
MBP
N-terminal maltose binding protein tag
1061
2165
tag
His
8xHis tag
2210
2234
trxn termination sequence
rrnB
rrnB transcription terminator
15
111
