Gene:
Gene Symbol:
PAS_chr3_0535
Gene Name:
Sphingolipid alpha-hydroxylase
Original Clone ID:
SCS7-11_pSGP46
Keyword:
None
Species:
Pichia pastoris
Type:
cDNA
Vector Name:
pSGP46
Format:
FUSION
Source:
Membrane Protein Structural Biology Consortium
Description:
None
Comments:
None
Mutation / Discrepancy:
No / Yes
Publications:
None
Authors:
University of Rochester
PSI:Biology
Membrane Protein Structural Biology Consortium
PSI:Biology
Membrane Protein Structural Biology Consortium
PpCD00586000
Price:
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AVAILABLE
No restriction
Special MTA: PSI
Special MTA: PSI
Insert sequence: 1149nts
Open reading frame: 40 to 1188
ACGGAATTTCCGACACCTCTCTTACACAAATATACCAAAATGACATCAAATAAGCCATTA
CCGCTCTTTGCCTCTGCTGATGTTCAGAAACACGTGAGTCCAAAGGATTGTTGGGTTACG
CTATATCATAGGAAGGTTTACAACGTTACAGAGTTTTTGGAGGAGCATCCGGCTGGAGAC
GACTTGATTTTGGAGTATGCTGGGAAGGATATCACACAGATTATGGGTGATCAGCTAAGT
CATATTCACTCCGAATCTGCTTATGAAATGTTGGATGAAGACATGTTGGTCGGATATTTG
GCAACACCGGAAGAAGAAAGGGAATTACTTTCCAATAAAGATCATGAGGTCGAAGTGCAG
CTAGACTCCAAGGGAAATGTAGTTGAAGTCGATATTGACCTAACTGAGTTTGGTGAACTT
CCGGCTGAGGGTTTACTCTCGTTGAAAACAGACTATGAACACGACTTGGTTAAACACCAA
TTTTTGGACTTGAACAGGCCTTTGTTGATGCAAGTTTTACGTGGATCATGGACTAAGGAG
TTTTATTTGGACCAAGTTCATCGTCCTAGACATTACGGAAAGGGTTCTGCTCCTCTTTTT
GGAAACTTTTTGGAACCTCTGAGTATGACAGCTTGGTGGATTGTACCCATGGTATGGCTT
CCTGTGAATTTCTATTTTTTCTACATTGGATTCACCAATCAAAACAAATTAGTCGCCATG
GCTTTTTGGCTTTTGGGTCTATTTGTTTGGACCTTCTTGGAATATGCTTTGCATAGATTT
TTGTTCCACTTGGACTACTATCTTCCAGAGAATCAAATTGCATTTACCATTCATTTCTTA
TTGCATGGGATACACCACTATTTACCAATGGATAAATACAGATTGGTGATGCCACCTACA
CTTTTCATTGTACTTTGCTACCCAATCAAGACGCTCGTCTTTTCTGTTCTACCATATTAC
ATGGCTTGTTCTGGATTTGCAGGTGGATTCCTGGGCTATATCATGTATGATGTCACTCAT
TACGTTCTGCATCACTCCAAGCTGCCTCGTTATTTCCAAGAGTTGAAGAAATATCATTTG
GAACATCACTACAAGAATTACGAGTTAGGCTTTGGTGTCACTTCCAAATTCTGGGACAAA
GTCTTTGGGACTTATCTGGGTCCAGACGATGTGTATCAGAAGACAAATAGTGGTTTGGAG
GTTTTGTTTCAAGGTCCAAACGGTGCTCCATCGGCCGTG
Protein sequence predicted based on nucleotide sequence
MTSNKPLPLFASADVQKHVSPKDCWVTLYHRKVYNVTEFLEEHPAGDDLILEYAGKDITQ IMGDQLSHIHSESAYEMLDEDMLVGYLATPEEERELLSNKDHEVEVQLDSKGNVVEVDID LTEFGELPAEGLLSLKTDYEHDLVKHQFLDLNRPLLMQVLRGSWTKEFYLDQVHRPRHYG KGSAPLFGNFLEPLSMTAWWIVPMVWLPVNFYFFYIGFTNQNKLVAMAFWLLGLFVWTFL EYALHRFLFHLDYYLPENQIAFTIHFLLHGIHHYLPMDKYRLVMPPTLFIVLCYPIKTLV FSVLPYYMACSGFAGGFLGYIMYDVTHYVLHHSKLPRYFQELKKYHLEHHYKNYELGFGV TSKFWDKVFGTYLGPDDVYQKTN
Coding Sequence Details
Insert Sequence Verified?:
Y
Verification Method:
Sequence Verification
Mutations:
None
Discrepancy:
g51a,K17K;g61c,A21P;
c90t,H30H;a93g,R31R;
t105c,N35N;c117t,F39F;
a126g,E42E;c171t,D57D;
g192t,E64D;t210c,H70H;
t213c,S71S;a219t,S73S;
g228a,E76E;a249g,L83L;
c259t,L87L;g267a,T89T;
a294c,S98S;a309g,E103E;
t312c,V104V;a333g,K111K;
g348a,E116E;t351c,V117V;
c354t,D118D;c363a,L121L;
a384g,P128P;a390g,E130E;
g408a,K136K;g435a,K145K
5' Linker Sequence:
None
3' Linker Sequence:
None
Recommended Growth Condition:
Distributed in bacterial strain:
DH5-alpha T1 phage resistant
Antibiotic Selection:
Host Type:
yeast
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:
SolubleProtein: Not_Applicable
ProteinExpressed: Protein_Confirmed
ProteinPurified: Tested_Not_Purified
Find experimental protocols in MPSBC-Scs7pPp
Recommended expression in:
Not Applicable
Vector Name:
pSGP46
Synonyms:
''
Sequencing Primer:
Forward:
pSGP-Forward
Reverse:
pSGP-Reverse
Description:
Yeast expression vector with Adh2 promoter, adds C-terminal 3C-ZZ-10XHis tag; ampicillin resistance (bacteria); uracil resistance (yeast); ligation independent cloning (LIC).
Comments:
S. cerevisiae expression strain
Size (bp):
6675
Parent Vector:
None
Properties:
ligation independent cloning (LIC), with tag/fusion/marker, yeast expression
Author Name:
Mark Dumont
Kathleen Clark
Center for High Throughput Structural Biology
PSI
Kathleen Clark
Center for High Throughput Structural Biology
PSI
Publications:
PMID:
20045057
Title: Purification of transmembrane proteins from Saccharomyces cerevisiae for X-ray crystallography
Vector Map:
Vector Sequence:
Sequence:
Vector Features:
Type
Name
Description
Start
End
ligation independent cloning
LIC
Ligation independent cloning site
0
0
promoter
Ura3 promoter
Ura3 promoter
6360
6389
protease cleavage site
3C
Rhinovirus 3C protease cleavage site
4233
4256
selectable marker
AmpR
Ampicillin resistance gene
1742
2401
selectable marker
Ura3
Ura3 nutritional marker (yeast)
5471
6274
tag
His
10x His tag
4653
4682
