Gene:
Gene Symbol:
VC0558
Gene Name:
hemolysin
Original Clone ID:
GE.349.1.69_GO.8405
Keyword:
None
Species:
Vibrio cholerae O1 biovar El Tor str. N16961
Type:
cDNA
Vector Name:
pNYCOMPS-LIC-FH10T+ (N term)
Format:
CLOSED
Source:
New York Consortium on Membrane Protein Structure
Description:
None
Comments:
None
Mutation / Discrepancy:
No / No
Publications:
None
Authors:
PSI
New York Consortium on Membrane Protein Structure
New York Consortium on Membrane Protein Structure
VcCD00340435
Price:
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AVAILABLE
No restriction
Special MTA: PSI
Special MTA: PSI
Insert sequence: 1281nts
Open reading frame: 1 to 1281
ATGGACGACATATCAACGGGTATCTTATTTGCGCTACTCGCGTGTCTCATCATTATTTCC
GCCTATTTCTCCGGTTCTGAAACCGGCATGATGGCTCTCAACCGTTACCGTTTAAAGCAT
TTGGCCAACAGCGGCCATAAAGGTGCTAAACGTGTTGAAAAGCTACTGGATCGTCCAGAT
CGTTTGATTGGCTTAATCCTGATTGGTAACAACCTCGTCAATATCCTCGCCTCTGCGATT
GCCACCATTATCGGTATGCGCCTGTATGGCGATTTAGGTGTCGCAATTGCCACGGGGGCC
TTAACTTTAGTGATTCTGGTGTTTGCCGAAGTCACGCCTAAAACTCTAGCCGCGCTTTAC
CCAGAGCGAGTCTCTTACGCCAGCAGCGTGTTACTGACGATTTTGATGAAGCTGCTTTCA
CCTTTGGTGCTGTTCGTTAACTTGATGACCAATGGCTTGATCCGCATTCTCGGCATCTCA
CCGAAACATGGCAAAGGCGATCATTTAAGCTCAGAAGAGTTACGTACTGTGGTTAATGAA
GCGGGAGGACTCATCCCTCGCCGCCACCAAGATATGCTGCTGTCGATCCTCGATTTAGAA
CACGTCACCGTCAACGACATCATGATCCCGCGTAATGAGATAACGGGGATCAATATCAAT
GATGATTGGAAGTCGATTGTGCGTCAGTTAGTCCATTCACCACATGGGCGAGTCGTACTG
TACCGTGACAAGATTGACGAAGTGGTCGGGATTTTGCGTTTACGTGAAGCCTCACGCTTT
ATGCTTGAGAAGAACGACTTTAACAAAGAGATGCTGCTGCGCGCTGCGGATGAGATTTAC
TTTATCCCTGAAGGTACACCGCTTAACGTTCAGCTGCTCAAATTCCAACGTAATAAAGAA
CGCATTGGTTTAATCGTGGATGAATACGGTGAAATCATTGGTTTAACCACGCTCGAAGAT
ATTTTGGAAGAGATTGTCGGTGAATTTACCACATCCATGTCTCCAAGCTTGGCCGATGAA
ATTACTCCACAAGGTGATGGCAGTTTCTTGATTGAAGGCAGCGCCAATATTCGTGACATT
AACAAGAGCCTGAAATGGAAGCTTCCTACCGATGGACCACGAACCTTAAACGGTTTGATT
TTGGAGCACCTTGAAGAGATCCCAGCGAGCCATCTGAGTGTGAAAGTGGCACAGCACAAA
ATGGAAATCCTCGAGCTGGAAGAGAATCGGATTAAGTTGGTCAAAGTTTACCCAAGAAAA
ACCAAAGTCAGCTTGGTTTAA
Protein sequence predicted based on nucleotide sequence
MDDISTGILFALLACLIIISAYFSGSETGMMALNRYRLKHLANSGHKGAKRVEKLLDRPD RLIGLILIGNNLVNILASAIATIIGMRLYGDLGVAIATGALTLVILVFAEVTPKTLAALY PERVSYASSVLLTILMKLLSPLVLFVNLMTNGLIRILGISPKHGKGDHLSSEELRTVVNE AGGLIPRRHQDMLLSILDLEHVTVNDIMIPRNEITGININDDWKSIVRQLVHSPHGRVVL YRDKIDEVVGILRLREASRFMLEKNDFNKEMLLRAADEIYFIPEGTPLNVQLLKFQRNKE RIGLIVDEYGEIIGLTTLEDILEEIVGEFTTSMSPSLADEITPQGDGSFLIEGSANIRDI NKSLKWKLPTDGPRTLNGLILEHLEEIPASHLSVKVAQHKMEILELEENRIKLVKVYPRK TKVSLV*
Coding Sequence Details
Insert Sequence Verified?:
Y
Verification Method:
Sequence Verification
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:
SolubleProtein: Not_Applicable
ProteinExpressed: Tested_Not_Found
ProteinPurified: Tested_Not_Purified
Find experimental protocols in GO.8405
Recommended expression in:
Not Applicable
Vector Name:
pNYCOMPS-LIC-FH10T+ (N term)
Synonyms:
''
Sequencing Primer:
Forward:
T7
Reverse:
T7 terminator
Description:
Bacterial expression vector; T7 promoter; N terminal Flag epitope and 10X His tags; tags can be removed by TEV protease; kanamycin resistance; ligation independent cloning.
Comments:
None
Size (bp):
6984
Parent Vector:
None
Properties:
bacterial expression, ligation independent cloning (LIC), with tag/fusion/marker
Author Name:
New York Consortium on Membrane Protein Structure
New York Structural Biology Center
New York Structural Biology Center
Publications:
PMID:
20394823
Title: High-throughput expression and purification of membrane proteins.
Vector Map:
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Vector Sequence:
Sequence:
Vector Features:
Type
Name
Description
Start
End
bacterial origin
ColE
ColE1 origin
1583
2265
death cassette
ccdB
ccdB death cassette (removed in plasmids containing an insert)
5393
5602
promoter
T7
T7 promoter
5146
5165
protease cleavage site
TEV
TEV protease cleavage site
5290
5310
repressor protein gene
LacI
Lac inhibitor
3677
4693
ribosome binding site
RBS
Ribosome Binding Site
5225
5230
selectable marker
KanR
kanamycin resistance
725
1537
selectable marker
CmR
Chloramphenicol resistance gene
6040
6696
tag
Flag
Flag epitope tag; N terminal
5236
5259
tag
His
10xHis tag; N terminal
5260
5289
trxn regulatory element
lac operator
lac operator
5165
5187
trxn termination sequence
T7 term
T7 terminator
91
137
