Gene:
Gene Symbol:
Gene Name:
Original Clone ID:
2501.1.08_GO.74368
Keyword:
partial cds
Species:
Saccharomyces cerevisiae
Type:
cDNA
Vector Name:
pVP16
Format:
CLOSED
Source:
Center for Eukaryotic Structural Genomics
Description:
None
Comments:
None
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:
PSI
Center for Eukaryotic Structural Genomics
Center for Eukaryotic Structural Genomics
ScCD00383577
Price:
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AVAILABLE
No restriction
Special MTA: PSI
Special MTA: PSI
Insert sequence: 897nts
Open reading frame: 28 to 924
ACAAGTTTGTACAAAAAAGCAGGCTCCGAAAACCTGTACTTCCAGTCCATGGAGTATGGA
CATCACGCTAGACCAGATTCAAAACGACCATTGGATGAAGGATCACCAGCAGCCGCAGGA
TTAACTTCTAAGAAGGCGAACGAAGCTTTAACAAGAAATAGAGAATTAACAACTGTATTA
GTCAAAAATTTGCCTAAGAGCTATAATCAAAACAAAGTCTATAAGTACTTCAAACATTGT
GGGCCCATTATACACGTTGATGTTGCGGATTCGCTGAAGAAGAACTTTCGTTTTGCACGT
ATTGAATTTGCCAGGTATGATGGAGCCCTCGCTGCAATAACCAAAACACACAAAGTTGTA
GGTCAGAATGAAATTATAGTATCTCATTTAACAGAATGCACATTATGGATGACGAATTTC
CCCCCAAGTTATACTCAAAGAAATATTAGAGATCTATTGCAAGATATTAACGTTGTTGCA
CTCAGTATACGGCTTCCCAGTTTACGATTCAATACAAGCAGAAGGTTCGCTTACATCGAT
GTTACTTCTAAAGAGGATGCAAGATATTGTGTAGAAAAATTGAATGGACTCAAAATAGAA
GGTTATACTTTAGTTACTAAAGTTTCCAATCCGCTGGAAAAGTCGAAACGGACCGATTCT
GCCACACTAGAGGGGCGAGAGATTATGATACGAAACCTAAGTACAGAATTGCTCGACGAA
AACCTTCTGAGGGAATCTTTTGAGGGATTTGGTTCCATTGAAAAAATCAACATACCTGCT
GGACAGAAAGAGCACAGTTTCAATAACTGTTGTGCATTTATGGTTTTTGAAAATAAAGAT
TCTGCTGAAAGGGCACTTCAAATGAATAGAAGTTTGTTAGGTAATAGGGAAATTTCCGTT
TCCTTGGCTGATAAAAAACCATAGGACC
Protein sequence predicted based on nucleotide sequence
ENLYFQSMEYGHHARPDSKRPLDEGSPAAAGLTSKKANEALTRNRELTTVLVKNLPKSYN QNKVYKYFKHCGPIIHVDVADSLKKNFRFARIEFARYDGALAAITKTHKVVGQNEIIVSH LTECTLWMTNFPPSYTQRNIRDLLQDINVVALSIRLPSLRFNTSRRFAYIDVTSKEDARY CVEKLNGLKIEGYTLVTKVSNPLEKSKRTDSATLEGREIMIRNLSTELLDENLLRESFEG FGSIEKINIPAGQKEHSFNNCCAFMVFENKDSAERALQMNRSLLGNREISVSLADKKP*
Coding Sequence Details
Insert Sequence Verified?:
Y
Verification Method:
Sequence Verification
Mutations:
None
Discrepancy:
del@19,3
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:
ampicillin
Bacterial Selection Condition:
100 ug/mL ampicillin
Growth Condition:
Growth with the single antibiotic in LB at 37 degrees is recommended.
Comments:
Commonly used conditions for ampicillin resistant plasmid clones.
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:
pVP16
Synonyms:
''
Sequencing Primer:
Forward:
MBP Forward
Reverse:
pQE Reverse
Description:
Bacterial (E. coli) expression, adds N-terminal 8xHis and MBP, adds TEV site on 5p end of insert; ampicillin resistance in bacteria; recombinational cloning; Gateway cloning into this vector will remove the Gateway ccdB death casette.
Comments:
TEV protease cleavage site is "partial" and not functional. The TEV cleavage site must be added to the insert by PCR, which allows amino acids encoded by the attB1 site to be cut off after protein expression. Gateway cloning into this vector will remove the Gateway cassette
Size (bp):
7631
Parent Vector:
None
Properties:
Gateway, acceptor (destination), bacterial expression, recombinational cloning, with tag/fusion/marker
Author Name:
Center for Eukaryotic Structural Genomics
Publications:
PMID:
15750721
Title: Results from high-throughput DNA cloning of Arabidopsis thaliana target genes using site-specific recombination
Vector Map:
Vector Sequence:
Sequence:
Vector Features:
Type
Name
Description
Start
End
bacterial origin
ori
ColE1-type bacterial origin of replication
6471
5789
negative selection marker
ccdB
ccdB death cassette gene (lost in with-insert form)
2643
2852
promoter
T5
T5 promoter
5
54
recombination site
AttR1
AttR1 recombination site
1316
1440
recombination site
AttR2
AttR2 recombination site
3020
2896
repressor protein gene
LacI
LacI coding region
4132
5211
selectable marker
CmR
chloramphenicol resistance gene (CmR) (Lost in with-insert form)
1549
2205
selectable marker
AmpR
ampicillin resistance gene
7228
6569
tag
MBP
N-terminal MBP
148
1246
tag
His
N-terminal 8xHis tag
128
145
