DNASU Plasmid | PFA4 (S. cerevisiae )<br>In pSGP47 (His-tagged yeast expression vector)

Explanation of Terms

Gene:

Gene Symbol: PFA4

Gene Name: Pfa4p

Original Clone ID:

YOL003C_pSGP47

Keyword:

None

Species:

Saccharomyces cerevisiae

Type:

cDNA

Vector Name:

pSGP47

Format:

FUSION

Source:

Membrane Protein Structural Biology Consortium

Description:

None

Comments:

None

Mutation / Discrepancy:

No / No

Publications:

None

Authors:

University of Rochester
PSI:Biology
Membrane Protein Structural Biology Consortium

ScCD00586006

Price:

AVAILABLE

No restriction
Special MTA: PSI

Insert Sequence
Protein Sequence

Insert sequence: 1134nts

Open reading frame: 39 to 1172

CGGAAATTCCGAACACCTTCTTACACAAATATACCAAAATGCCAGTAAAGTTAAGGTGGC
CTTGGTTAGGAATTGCTATACCAACGTTTCTGATTTCATTTATAGGCTATGGTGCACACT
ACTTTATTTTATCCAATTTCTTGTCAGTTCCGAAACAAATAACATTTGAGTTCTGTTTGT
CAATGATATGGCTCTCCTATTACTTGGCCATCTGCACAAATCCAGGAAGGCCTTTGCCCA
ACTACAAACCACCCCCTGATATCTGGCGGAATTTCTGTAAGAAGTGCCAAAGTTATAAGC
CAGAACGTTCACATCATTGTAAAACCTGCAATCAATGCGTCCTAATGATGGATCATCATT
GTCCGTGGACGATGAATTGCGTCGGCTTTGCAAACTATCCGCATTTCCTTAGATTCTTAT
TTTGGATCATAGTGACGACTAGTGTTTTATTTTGCATACAAGCTAAGCGGATATATTTTA
TTTGGCAACAAAGGCACTTGCCTGGCTACTTCTTCAAAAAATCAGAACTGATTTTCTTAA
CAATATCGAGTCCTTTAAATTCTTTTGTCCTACTAACGATAACAATTCTTTTCCTACGAT
GTTTATTCAATCAAATCTTGAATGGAAGATCTCAAATTGAGTCGTGGGATATGGATCGGC
TAGAATCTTTGTTCAACTCCGGAAGATTAACTCAGAAATTAATTGATAATACTTGGAGAA
TATATCCAGAGTCGAGGTCGTTTCAAAATAAAAAGGATGCTGAGGAACACTTGACAAAAA
AAAGGCCACGTTTTGATGAACTTGTAAATTTCCCTTACGATTTTGATTTGTACACTAATG
CATTACTGTACCTTGGTCCCATTCATTTATGGTTATGGCCATATGGTGTTCCTACGGGAG
ATGGGAACAATTTTCCAAAAAACGGTATTTCAAAATATGAGGCTAACTCCTCATTGGAAG
ACCACATCCTTTCACTTCCCTGGCCACCAGATGGAGGTAAAACAAACACCGTCTTCAATC
ATGGCAGCTCAACAATAGAGATGCGTAATGAAAGTGGCGAACAGCTAATAAGGACCAGAC
TTCCTCAAAATGGCAGACATGCTTCTCGTGAGAAGTGGTACAACGACTGGGGTGAATCAC
TAGACGATTTTGGAGTGGATGTTGATATGGAAAGTGGTTTGGAGGTTTTGTTTCAAGGTC
CAAACGGTGCTAGAGGTTCCCAT

Protein sequence predicted based on nucleotide sequence

MPVKLRWPWLGIAIPTFLISFIGYGAHYFILSNFLSVPKQITFEFCLSMIWLSYYLAICT
NPGRPLPNYKPPPDIWRNFCKKCQSYKPERSHHCKTCNQCVLMMDHHCPWTMNCVGFANY
PHFLRFLFWIIVTTSVLFCIQAKRIYFIWQQRHLPGYFFKKSELIFLTISSPLNSFVLLT
ITILFLRCLFNQILNGRSQIESWDMDRLESLFNSGRLTQKLIDNTWRIYPESRSFQNKKD
AEEHLTKKRPRFDELVNFPYDFDLYTNALLYLGPIHLWLWPYGVPTGDGNNFPKNGISKY
EANSSLEDHILSLPWPPDGGKTNTVFNHGSSTIEMRNESGEQLIRTRLPQNGRHASREKW
YNDWGESLDDFGVDVDME

Coding Sequence Details

Insert Sequence Verified?:

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: 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: Protein_Purified

Find experimental protocols in MPSBC-Pfa4p

Recommended expression in:

Not Applicable

Vector Name:

pSGP47

Synonyms:

Sequencing Primer:

Forward: pSGP-Forward

Reverse: pSGP-Reverse

Description:

Yeast expression vector with Adh2 promoter, adds 3C protease cleavage site and 10xHis tag; ampicillin resistance (bacteria); uracil (yeast); ligation independent cloning (LIC).

Comments:

S. cerevisiae expression strain

Size (bp):

6296

Parent Vector:

None

Properties:

ligation independent cloning (LIC), with tag/fusion/marker, yeast expression

Author Name:

Mark Dumont
Nadia Fedoriw
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:

GACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGTTGCTTCGTAGACACGAAGTAAAACATCTTGTTTTTACGTTGCGCTCTCGCGATTAAAAAGTTTGTTTCTTAGACTCGACGTAAAAATGTCTTGTCTTTACGTTGCGCTTTCGCGATAAAATGGTTGCTTCTTAGACACGAAGTAAAAACATTTTGTTTTTACGTTGCGCTCTCGCGATTAAAAAGTTTGTTTCTTAGACTCGACGTAAAAATGTCTTGTCTTTACGTTGCGCTCTCGCGATAAAATGGTTGTTTCTTAGATATGAAGAAAAAACAAGATGTTTTTACGTAGGGCTCTCGCGATAAAAAGATTGTTTCGTAGAATCTAATGAAAAAAAGAGGAAACACGCGAGATATTACGTCAGAGAACTATTGAAAAACGTGACATCCAGGCAATTCCAATCTTCTTCCGATGAAACCACAGATAAAAGAGAAGGTATTTTTTTCGGACTGAGGTGAAGGGCGCAAATGACTAATGATCGCTTCGACGCCCACGTAAAAAAGTTCTATTTCCGTAGGGGCTAATATAAGATATGGCTACACCTAACGCGTATGAAACACTTGTCTTTCACTATCGCAACTACTAAGAAGTAACCAGTCTTTTAATACTTGCCAAAGAAGATAAAACAGAGATATATGATGCATATCCTTTACAAATGTAAAAGCATAACAAAAGCTAAGTGAGATACTTATCAAGAATGATGTTAAAAAAACAGATTTCTCATTATGATCTCTATTTGTATTTTTTACATCTCCAGCTCAAATCTACGTTCAAGTTCCTCGCTTTCCACCTACCCATCCAATATATCCCTATATCGTGTCTCTATATATCGTTTCTCTATGAAAACTCGTTACAAACACCTTCGCCATAAGCGTTATAAAATCATCGAGCAATGTCAGGCCACGCAAAAACCAAAAAACTTTCACGCAGAAGTCTCGCGAAAACCAAAAGTTTTCGCGAGACTTCAAGGATATGAAAGATCTCTTATCCTTGAAGCCTTATCCTTGAAGTTTCGCAAAGGCTTTTGCTCGCGAAGGCTTTTACGTTGCGCTCGACGCGTGTATGTCGAGTGACAAGTGCAGCGTGGATATAGACGCACAACGGACATATATATATATGTACTCTTCTTGCCGTATCACGCACAAATACGAATTTACGCATGAATATACGCAGATAAATACATCCTACTTTCCATCAGATCATGGAGGACACTATAATAGGGTAAGGTACGCCCCATAGCATACGAAGGAAGTCGTGATGGGAAATCGACAAGATATACGACGGTGAGGAGTTAACCTAATCAGAGTAGGAAGTTACGATAGTAAAGGAAACTATAACCTAGTATACAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTACCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCGCGCAATTAACCCTCACTAAAGGGAACAAAAGCTGGAGCTCAAAACGTAGGGGCAAACAAACGGAAAAATCGTTTCTCAAATTTTCTGATGCCAAGAACTCTAACCAGTCTTATCTAAAAATTGCCTTATGATCCGTCTCTCCGGTTACAGCCTGTGTAACTGATTAATCCTGCCTTTCTAATCACCATTCTAATGTTTTAATTAAGGGATTTTGTCTTCATTAACGGCTTTCGCTCATAAAAATGTTATGACGTTTTGCCCGCAGGCGGGAAACCATCCACTTCACGAGACTGATCTCCTCTGCCGGAACACCGGGCATCTCCAACTTATAAGTTGGAGAAATAAGAGAATTTCAGATTGAGAGAATGAAAAAAAAAAAAAAAAAAAAGGCAGAGGAGAGCATAGAAATGGGGTTCACTTTTTGGTAAAGCTATAGCATGCCTATCACATATAAATAGAGTGCCAGTAGCGACTTTTTTCACACTCGAAATACTCTTACTACTGCTCTCTTGTTGTTTTTATCACTTCTTGTTTCTTCTTGGTAAATAGAATATCAAGCTACAAAAAGCATACAATCAACTATCAACTATTAACTATATCGTAATACGAATTCGAACACCTTCTTACAACAAATATACCAAAATTGTGGTTTGGAGGTTTTGTTTCAAGGTCCAAACGGTGCTAGAGGTTCCCATCACCATCACCATCACCATCACCATCACTGAGTACCCAATTCGCCCTATAGTGAGTCGTATTACGCGCGCTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATAGGGTAATAACTGATATAATTAAATTGAAGCTCTAATTTGTGAGTTTAGTATACATGCATTTACTTATAATACAGTTTTTTAGTTTTGCTGGCCGCATCTTCTCAAATATGCTTCCCAGCCTGCTTTTCTGTAACGTTCACCCTCTACCTTAGCATCCCTTCCCTTTGCAAATAGTCCTCTTCCAACAATAATAATGTCAGATCCTGTAGAGACCACATCATCCACGGTTCTATACTGTTGACCCAATGCGTCTCCCTTGTCATCTAAACCCACACCGGGTGTCATAATCAACCAATCGTAACCTTCATCTCTTCCACCCATGTCTCTTTGAGCAATAAAGCCGATAACAAAATCTTTGTCGCTCTTCGCAATGTCAACAGTACCCTTAGTATATTCTCCAGTAGATAGGGAGCCCTTGCATGACAATTCTGCTAACATCAAAAGGCCTCTAGGTTCCTTTGTTACTTCTTCTGCCGCCTGCTTCAAACCGCTAACAATACCTGGGCCCACCACACCGTGTGCATTCGTAATGTCTGCCCATTCTGCTATTCTGTATACACCCGCAGAGTACTGCAATTTGACTGTATTACCAATGTCAGCAAATTTTCTGTCTTCGAAGAGTAAAAAATTGTACTTGGCGGATAATGCCTTTAGCGGCTTAACTGTGCCCTCCATGGAAAAATCAGTCAAGATATCCACATGTGTTTTTAGTAAACAAATTTTGGGACCTAATGCTTCAACTAACTCCAGTAATTCCTTGGTGGTACGAACATCCAATGAAGCACACAAGTTTGTTTGCTTTTCGTGCATGATATTAAATAGCTTGGCAGCAACAGGACTAGGATGAGTAGCAGCACGTTCCTTATATGTAGCTTTCGACATGATTTATCTTCGTTTCCTGCAGGTTTTTGTTCTGTGCAGTTGGGTTAAGAATACTGGGCAATTTCATGTTTCTTCAACACTACATATGCGTATATATACCAATCTAAGTCTGTGCTCCTTCCTTCGTTCTTCCTTCTGTTCGGAGATTACCGAATCAAAAAAATTTCAAAGAAACCGAAATCAAAAAAAAGAATAAAAAAAAAATGATGAATTGAATTGAAAAGCTGTGGTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCAT

Vector Features:

Type

Name

Description

Start

End

ligation independent cloning

LIC

Ligation independent cloning site

4220

4231

primer

sequencing primer: CAATCAACTATCAACTATTAACTA

4155

4178

promoter

Ura3 promoter

5981

6010

protease cleavage site

Rhinovirus 3C protease cleavage site

4233

4256

selectable marker

AmpR

ampicillan selectable marker

1742

2401

selectable marker

Ura3

URA3 nutritional marker

5092

5895

tag

His

10xHis tag

4275

4304

Gene

Gene Symbol :

PFA4

GENEID :

854159

Locus Tag :

YOL003C

GI :

6324571

GenBank Accession :

NM_001183257

Yeast Genome Database :

S000005363

Target GenBank:

NM_001183257

Protein

Structural Annotation Wiki :

MPSBC-Pfa4p

Clone

Cloning Information :

599372

Original Clone ID :

YOL003C_pSGP47

Experimental

TargetTrack - Experimental data :

MPSBC-Pfa4p

Organism

Species Specific ID:

854159

Genome

Chromosome :

PFA4 (S. cerevisiae )In pSGP47 (His-tagged yeast expression vector)

Coding Sequence Details

Recommended Growth Condition:

Vector Features:

Gene

Protein

Clone

Experimental

Organism

Genome

PFA4 (S. cerevisiae )
In pSGP47 (His-tagged yeast expression vector)