DNASU Plasmid | MCA0337 (M. capsulatus str. Bath )<br>In pET21_NESG (His-tagged bacterial expression vector)

Explanation of Terms

Gene:

Gene Symbol: MCA0337

Gene Name: diguanylate phosphodiesterase

Original Clone ID:

McR174A-21.1

Keyword:

full length protein truncation

Species:

Methylococcus capsulatus str. Bath

Type:

cDNA

Vector Name:

pET21_NESG

Format:

FUSION

Source:

Northeast Structural Genomics Consortium

Description:

None

Comments:

None

Mutation / Discrepancy:

No / Yes

Publications:

None

Authors:

PSI
Northeast Structural Genomics Consortium

McCD00385582

Price:

AVAILABLE

No restriction
Special MTA: PSI/NESG

Insert Sequence
Protein Sequence

Insert sequence: 765nts

Open reading frame: 47 to 811

CCCCTCTAGAAATAATTTTGTTTAACTTTAAGAAGGAGATATACATATGCAGTTCTTCTC
GCCGGAACTGAACGTCCAGGCCTACCGGCGGCAGGAAATCGAAAGGAACCTCCGCTGCGC
CCTGCTCCAGGGCGAACTCGGCCTGAGCTATGCGCCCCGGGTGGATTTGCGCAGCGGCAG
GATCGTCGCCCTGGAGGTGTGTCCCTTCTGGCTCAGTTCCGAGTTGGGAGACGTGCCGCC
GGACGACTTCGTTCCCGTGGCGGAGGAAACCGGTCTCATCGTGCAGATCGGTATCTGGGC
TTTTCGCACCGCCTGCGCTCATGCGGCGGTGTGGCGGACGCAAGGACTCGACGACCGCTG
TCTCTCTTTCCGGTTGTCGCAGCGGGAACTGGATGATCCCCAGTTCGGCGATAGGATCAT
GAGTTTGCTGGATGGCTGTGGCCTGGAACCGGAGAACCTGCAGTTCGAACTCTCGGAGCG
CATGCTGATGACGCATGCCGCACACACCCTCGCCGTGCTGCAGGAACTCAGCAACCGGGG
CATCCGCTTCTCGGTGGGGGACTTCGGCACGGGCAATGCGTCCTTGCGGCGGCTGCGGCG
GTTTCCGATCGATGAACTCAAGATCGATCTGTCATCCGTCCGGGACATCGAGCCGGATTC
GGACGATGCCACCGTCATCACCGCGATGATTGCCATGGCCAAGGGTCTGCGGATGGACGT
CGTCGCCGTCGGCGTGGAAACCCCGGAGCACGTGGCATTCCTGCTTGAGCGCGGGTGTCG
TCACGGGCAGGGCCGTCTGATCGGGGCGCGGCTCGAGCACCACCACCACCACCACTGAGA
TCCGGCTGCTAACAAAGCC

Protein sequence predicted based on nucleotide sequence

MQFFSPELNVQAYRRQEIERNLRCALLQGELGLSYAPRVDLRSGRIVALEVCPFWLSSEL
GDVPPDDFVPVAEETGLIVQIGIWAFRTACAHAAVWRTQGLDDRCLSFRLSQRELDDPQF
GDRIMSLLDGCGLEPENLQFELSERMLMTHAAHTLAVLQELSNRGIRFSVGDFGTGNASL
RRLRRFPIDELKIDLSSVRDIEPDSDDATVITAMIAMAKGLRMDVVAVGVETPEHVAFLL
ERGCRHGQGRLIGAR

Coding Sequence Details

Insert Sequence Verified?:

Verification Method:

Sequence Verification

Mutations:

None

Discrepancy:

c100a,R34S;a177g,E59E;
c178t,L60L;a205g,I69V;
c300a,G100G;c305a,A102D;
t396c,G132G;c429g,S143S;
g625a,A209T;a658g,S220G

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:

ProteinPurified: Not_Tested

ProteinExpressed: Protein_Confirmed

SolubleProtein: Tested_Not_Soluble

Find experimental protocols in McR174A

Recommended expression in:

Not Applicable

Vector Name:

pET21_NESG

Synonyms:

pET21, pET21-modified, pET21Cc, pET21_NESG

Sequencing Primer:

Forward: T7

Reverse: T7 terminator

Description:

Bacterial (E. coli) expression vector with T7 lac promoter, adds C-terminal 6xHis tag; ampicillin resistance; restriction enzyme cloning.

Comments:

Modified Novagen pET vector-common MCS

Size (bp):

5415

Parent Vector:

None

Properties:

bacterial expression, in vitro transcription, multiple cloning site, with tag/fusion/marker

Author Name:

Gaetano T. Montleione
Thomas B. Acton
Northeast Structural Genomics Consortium

Publications:

PMID: 15808222

Title: Robotic cloning and Protein Production Platform of the Northeast Structural Genomics Consortium

Vector Map:

Vector Sequence:

Sequence:

ATCCGGATATAGTTCCTCCTTTCAGCAAAAAACCCCTCAAGACCCGTTTAGAGGCCCCAAGGGGTTATGCTAGTTATTGCTCAGCGGTGGCAGCAGCCAACTCAGCTTCCTTTCGGGCTTTGTTAGCAGCCGGATCTCAGTGGTGGTGGTGGTGGTGCTCGAGTGCGGCCGCAAGCTTGTCGACGGAGCTCGAATTCGGATCCCCCGCCATATGTATATCTCCTTCTTAAAGTTAAACAAAATTATTTCTAGAGGGGAATTGTTATCCGCTCACAATTCCCCTATAGTGAGTCGTATTAATTTCGCGGGATCGAGATCTCGATCCTCTACGCCGGACGCATCGTGGCCGGCATCACCGGCGCCACAGGTGCGGTTGCTGGCGCCTATATCGCCGACATCACCGATGGGGAAGATCGGGCTCGCCACTTCGGGCTCATGAGCGCTTGTTTCGGCGTGGGTATGGTGGCAGGCCCCGTGGCCGGGGGACTGTTGGGCGCCATCTCCTTGCATGCACCATTCCTTGCGGCGGCGGTGCTCAACGGCCTCAACCTACTACTGGGCTGCTTCCTAATGCAGGAGTCGCATAAGGGAGAGCGTCGAGATCCCGGACACCATCGAATGGCGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAATGTGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTAAGTTAGCTCACTCATTAGGCACCGGGATCTCGACCGATGCCCTTGAGAGCCTTCAACCCAGTCAGCTCCTTCCGGTGGGCGCGGGGCATGACTATCGTCGCCGCACTTATGACTGTCTTCTTTATCATGCAACTCGTAGGACAGGTGCCGGCAGCGCTCTGGGTCATTTTCGGCGAGGACCGCTTTCGCTGGAGCGCGACGATGATCGGCCTGTCGCTTGCGGTATTCGGAATCTTGCACGCCCTCGCTCAAGCCTTCGTCACTGGTCCCGCCACCAAACGTTTCGGCGAGAAGCAGGCCATTATCGCCGGCATGGCGGCCCCACGGGTGCGCATGATCGTGCTCCTGTCGTTGAGGACCCGGCTAGGCTGGCGGGGTTGCCTTACTGGTTAGCAGAATGAATCACCGATACGCGAGCGAACGTGAAGCGACTGCTGCTGCAAAACGTCTGCGACCTGAGCAACAACATGAATGGTCTTCGGTTTCCGTGTTTCGTAAAGTCTGGAAACGCGGAAGTCAGCGCCCTGCACCATTATGTTCCGGATCTGCATCGCAGGATGCTGCTGGCTACCCTGTGGAACACCTACATCTGTATTAACGAAGCGCTGGCATTGACCCTGAGTGATTTTTCTCTGGTCCCGCCGCATCCATACCGCCAGTTGTTTACCCTCACAACGTTCCAGTAACCGGGCATGTTCATCATCAGTAACCCGTATCGTGAGCATCCTCTCTCGTTTCATCGGTATCATTACCCCCATGAACAGAAATCCCCCTTACACGGAGGCATCAGTGACCAAACAGGAAAAAACCGCCCTTAACATGGCCCGCTTTATCAGAAGCCAGACATTAACGCTTCTGGAGAAACTCAACGAGCTGGACGCGGATGAACAGGCAGACATCTGTGAATCGCTTCACGACCACGCTGATGAGCTTTACCGCAGCTGCCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATGCAGCTCCCGGAGACGGTCACAGCTTGTCTGTAAGCGGATGCCGGGAGCAGACAAGCCCGTCAGGGCGCGTCAGCGGGTGTTGGCGGGTGTCGGGGCGCAGCCATGACCCAGTCACGTAGCGATAGCGGAGTGTATACTGGCTTAACTATGCGGCATCAGAGCAGATTGTACTGAGAGTGCACCATATATGCGGTGTGAAATACCGCACAGATGCGTAAGGAGAAAATACCGCATCAGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTGCAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGAAATTGTAAACGTTAATATTTTGTTAAAATTCGCGTTAAATTTTTGTTAAATCAGCTCATTTTTTAACCAATAGGCCGAAATCGGCAAAATCCCTTATAAATCAAAAGAATAGACCGAGATAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTCCACTATTAAAGAACGTGGACTCCAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATGGCCCACTACGTGAACCATCACCCTAATCAAGTTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAATCGGAACCCTAAAGGGAGCCCCCGATTTAGAGCTTGACGGGGAAAGCCGGCGAACGTGGCGAGAAAGGAAGGGAAGAAAGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTGTAGCGGTCACGCTGCGCGTAACCACCACACCCGCCGCGCTTAATGCGCCGCTACAGGGCGCGTCCCATTCGCCA

Vector Features:

Type

Name

Description

Start

End

MCS

MCS (includes SphI, XbaI, BamHI, EcoRI, XhoI)

157

253

bacterial origin

ori

pBR322-type bacterial origin of replication

3180

3862

gene fragment

ROP

2580

2756

primer

T7 forward

T7 forward sequencing primer

283

298

primer

T7 reverse

T7 reverse sequencing primer

promoter

T7 promoter

280

299

repressor binding site

lacO

lacO regulatory sequence

258

280

repressor protein gene

LacI

752

1768

selectable marker

AmpR

ampicillin resistance gene

3960

4619

ssDNA origin

f1 ori

f1 origin

5081

5387

tag

His

C-terminal 6xHis tag

140

157

trxn termination sequence

T7 term

T7 termination sequence

Gene

Gene Symbol :

MCA0337

GENEID :

3102149

Locus Tag :

MCA0337

GI :

53802334

Target GenBank:

None

Protein

SMART domain :

EAL : Putative diguanylate phosphodiesterase

Structural Annotation Wiki :

TOPSAN

UniProt :

Q60BX6

UniProt :

Q60BX6_METCA

Clone

Cloning Information :

546174

HIP Master Clone ID :

456494

Original Clone ID :

McR174A-21.1

Experimental

TargetTrack - Experimental data :

McR174A

Organism

Species Specific ID:

3102149

Reagents

Labome :

MCA0337-cDNA

MCA0337 (M. capsulatus str. Bath )In pET21_NESG (His-tagged bacterial expression vector)

Coding Sequence Details

Recommended Growth Condition:

Vector Features:

Gene

Protein

Clone

Experimental

Organism

Reagents

MCA0337 (M. capsulatus str. Bath )
In pET21_NESG (His-tagged bacterial expression vector)