DNASU Plasmid | fdsB (C. sphaeroides 2.4.1 )<br>In pNIC28-Bsa4 (His-tagged bacterial expression vector)

Explanation of Terms

Gene:

Gene Symbol: fdsB

Gene Name: formate dehydrogenase beta subunit

Original Clone ID:

014635

Keyword:

None

Species:

Cereibacter sphaeroides 2.4.1

Type:

cDNA

Vector Name:

pNIC28-Bsa4

Format:

CLOSED

Source:

New York Structural Genomics Research Consortium

Description:

None

Comments:

None

Mutation / Discrepancy:

No / No

Publications:

None

Authors:

Albert Einstein College of Medicine
Steven Almo
PSI:Biology
New York Structural Genomics Research Consortium

RsCD00526564

Price:

AVAILABLE

No restriction
Special MTA: PSI

Insert Sequence
Protein Sequence

Insert sequence: 1527nts

Open reading frame: 12 to 1538

TCCAATCCATGAAGATCTACGTTCCCCGCGACGCGGCCGCCAAGGCGCTCGGCGCCGACG
AGGTGGCTGCCGCCATCGAGGCCGAAGCCGCGCGGCGCGGCCTGCCGCTGACGCTCGTGC
GCAACGGCTCGCGCGGGATGGTCTGGTTGGAACCTTTGGTCGAGCTTCAGACCGGGGCCG
GCCGCGTGGCCTTCGGCCCGGTCACGGCCGCCGATGTGCCGAGCCTGTTCGAGGCGGACT
GGAAGCGGCATCCGCTGGCGCTGGGTCTGACGGAAGAGCTGCCGTGGCTCATGATGCAGA
CGCGCCTCACCTTCGCCCGCTGCGGGATCACCGATCCGCTCTGCCTCGAGGAGTATGAGG
CGCACGGCGGGTTCGAGGGGCTTCACCGCGCTATCGCCATGAGCCGCGCCGAGATCCTGC
AGGAGGTGATCGATTCCGGCCTCCGCGGGCGAGGCGGCGCGGGCTTTCCCACCGGCATCA
AGTGGAAGACGGTTTCCGAGGCGGAGGCCGCGCAGAAATACATCGTCTGCAACGCGGACG
AGGGCGACAGCGGCACCTTCGCCGACCGGATGATCATGGAGGGCGACCCCTTCACCCTGA
TCGAGGGCATGACCATCGCCGGCCTCGCGACCGGGGCCACGCGCGGCTATGTCTACATGC
GCTCGGAATATCCCGATGCGGTCGAGGTGATGGGCGAGGCGGTGCGGATCGCCCGCGCCC
ACGGGCTGCTCGGGCCGTCGGTCATGGGCTCGGGCCGCGCCTTCGACATGGAGATCCGCG
TCGGCGCCGGGGCCTATGTCTGCGGCGAGGAGACCTCGCTTCTGAACTCGCTCGAGGGCA
AGCGCGGCGTGGTGCGCGCCAAGCCGCCGCTTCCGGCGCTCGAGGGATTCCTCGGCAAGC
CCACGGTCGTGAACAACGTGATCTCGCTGGCCTCGATCCCGGTGATCTTCCAGAAGGGCG
CCGCCTACTACCGCGACTTCGGCCTCGGCCGGTCGCGCGGCACCATCCCGCTGCAGATCG
CGGGCAATGTGAAGCGCGGCGGCCTGTTCGAGATCGCCTTCGGCATGAGCCTCGGCCAGA
TCGTCGATGACATCGCGGGCGGCACGGCCTCGGGTCGGCCGGTCAAGGCCGTGCAGGTGG
GCGGGCCGCTCGGCGCCTACTTCCCGCGCGCCCTGTTCGATACGCCCTTCTGCTACGAGG
AATTCGCCGGAGCCGAGGGCCTCGTGGGCCATGCGGGCATCGTCGTCTTCGACGACACGG
CAGACATGCTGAAGCAGGCGCGCTTCGCGATGGAGTTCTGCGCCATCGAGAGCTGCGGGA
AATGCACCCCCTGCCGCATCGGCGCGGTGCGGGGCATGGAGACGGTCGACCGCATCGCCC
GCGGCGACGCGGCCGCGATCCCGCTGCTCACGGATCTGTGCGCCACGATGAAATCGGGAT
CGCTCTGCGCGCTGGGCGGTTTCACCCCCTACCCGGTGATGAGCGCCCTTACCCACTTCC
CGGACGATTTCGCCCCGATGAAGGAGGCAGCAGAATAACAGTAAAGGTGGA

Protein sequence predicted based on nucleotide sequence

KIYVPRDAAAKALGADEVAAAIEAEAARRGLPLTLVRNGSRGMVWLEPLVELQTGAGRVA
FGPVTAADVPSLFEADWKRHPLALGLTEELPWLMMQTRLTFARCGITDPLCLEEYEAHGG
FEGLHRAIAMSRAEILQEVIDSGLRGRGGAGFPTGIKWKTVSEAEAAQKYIVCNADEGDS
GTFADRMIMEGDPFTLIEGMTIAGLATGATRGYVYMRSEYPDAVEVMGEAVRIARAHGLL
GPSVMGSGRAFDMEIRVGAGAYVCGEETSLLNSLEGKRGVVRAKPPLPALEGFLGKPTVV
NNVISLASIPVIFQKGAAYYRDFGLGRSRGTIPLQIAGNVKRGGLFEIAFGMSLGQIVDD
IAGGTASGRPVKAVQVGGPLGAYFPRALFDTPFCYEEFAGAEGLVGHAGIVVFDDTADML
KQARFAMEFCAIESCGKCTPCRIGAVRGMETVDRIARGDAAAIPLLTDLCATMKSGSLCA
LGGFTPYPVMSALTHFPDDFAPMKEAAE*

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: 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 NYSGRC-014635

Recommended expression in:

Not Applicable

Vector Name:

pNIC28-Bsa4

Synonyms:

pSEC-His

Sequencing Primer:

Forward: pLICf

Reverse: pLIC rev

Description:

Bacterial expression vector with T7 promoter and N-terminal 6xHis tag with TEV protease cleavage site; kanamycin resistance in bacteria; ligation independent cloning

Comments:

pET expression vector with His6 tag in 22-aa N-terminal fusion peptide, with TEV protease cleavage site. Includes sites for LIC cloning, and a �stuffer� fragment that includes the SacB gene, allowing negative selection on 5% sucrose

Size (bp):

7284

Parent Vector:

None

Properties:

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

Author Name:

Opher Gileadi

Publications:

PMID: 20541610

Title: High-throughput production of human proteins for crystallization: the SGC experience

Vector Map:

Vector Sequence:

Sequence:

CTAGAAATAATTTTGTTTAACTTTAAGAAGGAGATATACATATGCACCATCATCATCATCATTCTTCTGGTGTAGATCTGGGTACCGAGAACCTGTACTTCCAATCCATGGAGACCGACGTCCACATATACCTGCCGTTCACTATTATTTAGTGAAATGAGATATTATGATATTTTCTGAATTGTGATTAAAAAGGCAACTTTATGCCCATGCAACAGAAACTATAAAAAATACAGAGAATGAAAAGAAACAGATAGATTTTTTAGTTCTTTAGGCCCGTAGTCTGCAAATCCTTTTATGATTTTCTATCAAACAAAAGAGGAAAATAGACCAGTTGCAATCCAAACGAGAGTCTAATAGAATGAGGTCGAAAAGTAAATCGCGCGGGTTTGTTACTGATAAAGCAGGCAAGACCTAAAATGTGTAAAGGGCAAAGTGTATACTTTGGCGTCACCCCTTACATATTTTAGGTCTTTTTTTATTGTGCGTAACTAACTTGCCATCTTCAAACAGGAGGGCTGGAAGAAGCAGACCGCTAACACAGTACATAAAAAAGGAGACATGAACGATGAACATCAAAAAGTTTGCAAAACAAGCAACAGTATTAACCTTTACTACCGCACTGCTGGCAGGAGGCGCAACTCAAGCGTTTGCGAAAGAAACGAACCAAAAGCCATATAAGGAAACATACGGCATTTCCCATATTACACGCCATGATATGCTGCAAATCCCTGAACAGCAAAAAAATGAAAAATATAAAGTTCCTGAGTTCGATTCGTCCACAATTAAAAATATCTCTTCTGCAAAAGGCCTGGACGTTTGGGACAGCTGGCCATTACAAAACACTGACGGCACTGTCGCAAACTATCACGGCTACCACATCGTCTTTGCATTAGCCGGAGATCCTAAAAATGCGGATGACACATCGATTTACATGTTCTATCAAAAAGTCGGCGAAACTTCTATTGACAGCTGGAAAAACGCTGGCCGCGTCTTTAAAGACAGCGACAAATTCGATGCAAATGATTCTATCCTAAAAGACCAAACACAAGAATGGTCAGGTTCAGCCACATTTACATCTGACGGAAAAATCCGTTTATTCTACACTGATTTCTCCGGTAAACATTACGGCAAACAAACACTGACAACTGCACAAGTTAACGTATCAGCATCAGACAGCTCTTTGAACATCAACGGTGTAGAGGATTATAAATCAATCTTTGACGGTGACGGAAAAACGTATCAAAATGTACAGCAGTTCATCGATGAAGGCAACTACAGCTCAGGCGACAACCATACGCTGAGAGATCCTCACTACGTAGAAGATAAAGGCCACAAATACTTAGTATTTGAAGCAAACACTGGAACTGAAGATGGCTACCAAGGCGAAGAATCTTTATTTAACAAAGCATACTATGGCAAAAGCACATCATTCTTCCGTCAAGAAAGTCAAAAACTTCTGCAAAGCGATAAAAAACGCACGGCTGAGTTAGCAAACGGCGCTCTCGGTATGATTGAGCTAAACGATGATTACACACTGAAAAAAGTGATGAAACCGCTGATTGCATCTAACACAGTAACAGATGAAATTGAACGCGCGAACGTCTTTAAAATGAACGGCAAATGGTACCTGTTCACTGACTCCCGCGGATCAAAAATGACGATTGACGGCATTACGTCTAACGATATTTACATGCTTGGTTATGTTTCTAATTCTTTAACTGGCCCATACAAGCCGCTGAACAAAACTGGCCTTGTGTTAAAAATGGATCTTGATCCTAACGATGTAACCTTTACTTACTCACACTTCGCTGTACCTCAAGCGAAAGGAAACAATGTCGTGATTACAAGCTATATGACAAACAGAGGATTCTACGCAGACAAACAATCAACGTTTGCGCCTAGCTTCCTGCTGAACATCAAAGGCAAGAAAACATCTGTTGTCAAAGACAGCATCCTTGAACAAGGACAATTAACAGTTAACAAATAAAAACGCAAAAGAAAATGCCGATATCCTATTGGCATTGACGGTCTCCAGTAAAGGTGGATACGGATCCGAATTCGAGCTCCGTCGACAAGCTTGCGGCCGCACTCGAGCACCACCACCACCACCACTGAGATCCGGCTGCTAACAAAGCCCGAAAGGAAGCTGAGTTGGCTGCTGCCACCGCTGAGCAATAACTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTGCTGAAAGGAGGAACTATATCCGGATTGGCGAATGGGACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAATTAATTCTTAGAAAAACTCATCGAGCATCAAATGAAACTGCAATTTATTCATATCAGGATTATCAATACCATATTTTTGAAAAAGCCGTTTCTGTAATGAAGGAGAAAACTCACCGAGGCAGTTCCATAGGATGGCAAGATCCTGGTATCGGTCTGCGATTCCGACTCGTCCAACATCAATACAACCTATTAATTTCCCCTCGTCAAAAATAAGGTTATCAAGTGAGAAATCACCATGAGTGACGACTGAATCCGGTGAGAATGGCAAAAGTTTATGCATTTCTTTCCAGACTTGTTCAACAGGCCAGCCATTACGCTCGTCATCAAAATCACTCGCATCAACCAAACCGTTATTCATTCGTGATTGCGCCTGAGCGAGACGAAATACGCGATCGCTGTTAAAAGGACAATTACAAACAGGAATCGAATGCAACCGGCGCAGGAACACTGCCAGCGCATCAACAATATTTTCACCTGAATCAGGATATTCTTCTAATACCTGGAATGCTGTTTTCCCGGGGATCGCAGTGGTGAGTAACCATGCATCATCAGGAGTACGGATAAAATGCTTGATGGTCGGAAGAGGCATAAATTCCGTCAGCCAGTTTAGTCTGACCATCTCATCTGTAACATCATTGGCAACGCTACCTTTGCCATGTTTCAGAAACAACTCTGGCGCATCGGGCTTCCCATACAATCGATAGATTGTCGCACCTGATTGCCCGACATTATCGCGAGCCCATTTATACCCATATAAATCAGCATCCATGTTGGAATTTAATCGCGGCCTAGAGCAAGACGTTTCCCGTTGAATATGGCTCATAACACCCCTTGTATTACTGTTTATGTAAGCAGACAGTTTTATTGTTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATATATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGTATACACTCCGCTATCGCTACGTGACTGGGTCATGGCTGCGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGGCAGCTGCGGTAAAGCTCATCAGCGTGGTCGTGAAGCGATTCACAGATGTCTGCCTGTTCATCCGCGTCCAGCTCGTTGAGTTTCTCCAGAAGCGTTAATGTCTGGCTTCTGATAAAGCGGGCCATGTTAAGGGCGGTTTTTTCCTGTTTGGTCACTGATGCCTCCGTGTAAGGGGGATTTCTGTTCATGGGGGTAATGATACCGATGAAACGAGAGAGGATGCTCACGATACGGGTTACTGATGATGAACATGCCCGGTTACTGGAACGTTGTGAGGGTAAACAACTGGCGGTATGGATGCGGCGGGACCAGAGAAAAATCACTCAGGGTCAATGCCAGCGCTTCGTTAATACAGATGTAGGTGTTCCACAGGGTAGCCAGCAGCATCCTGCGATGCAGATCCGGAACATAATGGTGCAGGGCGCTGACTTCCGCGTTTCCAGACTTTACGAAACACGGAAACCGAAGACCATTCATGTTGTTGCTCAGGTCGCAGACGTTTTGCAGCAGCAGTCGCTTCACGTTCGCTCGCGTATCGGTGATTCATTCTGCTAACCAGTAAGGCAACCCCGCCAGCCTAGCCGGGTCCTCAACGACAGGAGCACGATCATGCGCACCCGTGGGGCCGCCATGCCGGCGATAATGGCCTGCTTCTCGCCGAAACGTTTGGTGGCGGGACCAGTGACGAAGGCTTGAGCGAGGGCGTGCAAGATTCCGAATACCGCAAGCGACAGGCCGATCATCGTCGCGCTCCAGCGAAAGCGGTCCTCGCCGAAAATGACCCAGAGCGCTGCCGGCACCTGTCCTACGAGTTGCATGATAAAGAAGACAGTCATAAGTGCGGCGACGATAGTCATGCCCCGCGCCCACCGGAAGGAGCTGACTGGGTTGAAGGCTCTCAAGGGCATCGGTCGAGATCCCGGTGCCTAATGAGTGAGCTAACTTACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCGTGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCCAGGGTGGTTTTTCTTTTCACCAGTGAGACGGGCAACAGCTGATTGCCCTTCACCGCCTGGCCCTGAGAGAGTTGCAGCAAGCGGTCCACGCTGGTTTGCCCCAGCAGGCGAAAATCCTGTTTGATGGTGGTTAACGGCGGGATATAACATGAGCTGTCTTCGGTATCGTCGTATCCCACTACCGAGATATCCGCACCAACGCGCAGCCCGGACTCGGTAATGGCGCGCATTGCGCCCAGCGCCATCTGATCGTTGGCAACCAGCATCGCAGTGGGAACGATGCCCTCATTCAGCATTTGCATGGTTTGTTGAAAACCGGACATGGCACTCCAGTCGCCTTCCCGTTCCGCTATCGGCTGAATTTGATTGCGAGTGAGATATTTATGCCAGCCAGCCAGACGCAGACGCGCCGAGACAGAACTTAATGGGCCCGCTAACAGCGCGATTTGCTGGTGACCCAATGCGACCAGATGCTCCACGCCCAGTCGCGTACCGTCTTCATGGGAGAAAATAATACTGTTGATGGGTGTCTGGTCAGAGACATCAAGAAATAACGCCGGAACATTAGTGCAGGCAGCTTCCACAGCAATGGCATCCTGGTCATCCAGCGGATAGTTAATGATCAGCCCACTGACGCGTTGCGCGAGAAGATTGTGCACCGCCGCTTTACAGGCTTCGACGCCGCTTCGTTCTACCATCGACACCACCACGCTGGCACCCAGTTGATCGGCGCGAGATTTAATCGCCGCGACAATTTGCGACGGCGCGTGCAGGGCCAGACTGGAGGTGGCAACGCCAATCAGCAACGACTGTTTGCCCGCCAGTTGTTGTGCCACGCGGTTGGGAATGTAATTCAGCTCCGCCATCGCCGCTTCCACTTTTTCCCGCGTTTTCGCAGAAACGTGGCTGGCCTGGTTCACCACGCGGGAAACGGTCTGATAAGAGACACCGGCATACTCTGCGACATCGTATAACGTTACTGGTTTCACATTCACCACCCTGAATTGACTCTCTTCCGGGCGCTATCATGCCATACCGCGAAAGGTTTTGCGCCATTCGATGGTGTCCGGGATCTCGACGCTCTCCCTTATGCGACTCCTGCATTAGGAAGCAGCCCAGTAGTAGGTTGAGGCCGTTGAGCACCGCCGCCGCAAGGAATGGTGCATGCAAGGAGATGGCGCCCAACAGTCCCCCGGCCACGGGGCCTGCCACCATACCCACGCCGAAACAAGCGCTCATGAGCCCGAAGTGGCGAGCCCGATCTTCCCCATCGGTGATGTCGGCGATATAGGCGCCAGCAACCGCACCTGTGGCGCCGGTGATGCCGGCCACGATGCGTCCGGCGTAGAGGATCGAGATCTCGATCCCGCGAAATTAATACGACTCACTATAGGGGAATTGTGAGCGGATAACAATTCCCCT

Vector Features:

Type

Name

Description

Start

End

bacterial origin

ColE

ColE1 pBR322 origin

4338

bacterial origin

f1 origin

2266

2721

ligation independent cloning

5'LIC

110

ligation independent cloning

3'LIC

2036

2050

primer

pLICfor

pLIC forward sequencing primer TGTGAGCGGATAACAATTCC

7262

7282

primer

pLICrev

pLIC reverse sequencing primer AGCAGCCAACTCAGCTTCC

2145

2164

promoter

T7 promoter

7238

7254

protease cleavage site

TEV

TEV protease cleavage site

110

repressor binding site

lac operator

7257

7281

repressor protein gene

LacI

Lac repressor gene

5772

6851

selectable marker

KanR

kanamycin resistance

2817

3629

selectable marker

SacB

SacB gene - allows for negative selection on 5% sucrose

569

1987

tag

His

N-terminal 6xHis tag

trxn termination sequence

T7 term

T7 terminator

2183

2229

Gene

Gene Symbol :

fdsB

GENEID :

3720894

Locus Tag :

RSP_1079

GI :

77389078

GenBank Accession :

ABA80263.1

Target GenBank:

None

Protein

SMART domain :

NADH_4Fe-4S : NADH-ubiquinone oxidoreductase-F iron-sulfur binding region

Structural Annotation Wiki :

NYSGRC-014635

Clone

Cloning Information :

594944

Original Clone ID :

014635

Experimental

TargetTrack - Experimental data :

NYSGRC-014635

Organism

Species Specific ID:

3720894

Genome

Chromosome :

Reagents

Labome :

fdsB-cDNA

fdsB (C. sphaeroides 2.4.1 )In pNIC28-Bsa4 (His-tagged bacterial expression vector)

Coding Sequence Details

Recommended Growth Condition:

Vector Features:

Gene

Protein

Clone

Experimental

Organism

Genome

Reagents

fdsB (C. sphaeroides 2.4.1 )
In pNIC28-Bsa4 (His-tagged bacterial expression vector)