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Explanation of Terms


Gene: Gene Symbol:  None
Gene Name:  None
Sequence             
Original Clone ID: MBP-linker-MPR-TM
Keyword: There is a TEV cleavage site after the linker. The DNA sequence of the TEV cleavage site is: GAGAATCTTTATTTTCAGGGC; vector name: MBP-linker-MPR-TM; vector use: E. coli expression; selection marker: bacterial ampicillin; cloning type: Gatew
Species: Human immunodeficiency virus 1
Type: cDNA
Vector Name: pVP16               Format:  FUSION
Source: Center for Membrane Proteins in Infectious Diseases
Description: None
Comments: None
Mutation/
Discrepancy :
No / No
Publications: PMID: 26295457
Title: Biophysical Characterization of a Vaccine Candidate against HIV-1: The Transmembrane and Membrane Proximal Domains of HIV-1 gp41 as a Maltose Binding Protein Fusion
Authors: Zhen Gong
PSI:Biology
Center for Membrane Proteins in Infectious Diseases
Sasha M. Daskalova
HiCD00674886

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AVAILABLE
No restriction
Special MTA: PSI

 

Insert sequence: 177nts         Open reading frame : 1 to 177

ATGGGATCTCAAACTCAACAAGAGAAGAACGAGCAAGAGTTGTTGGAGTTGGATAAGTGG
GCTTCTTTGTGGAACTGGTTCGATATCACCAACTGGTTGTGGTACATTAAGATTTTCATT
ATGATTGTGGGAGGATTGATTGGATTGAGGATCATTTTCGCTGTGTTGTCTATGGTG


Coding Sequence Details
Insert Sequence Verified?: Y Verification Method: Sequence Verification
Mutations:
None
Discrepancy:
None
5' Linker Sequence:
TCGAGCTCGAACAACAACAACAATAACAATAACAACAACCTCGGGATCGA
CGAAAACCTGTACCTCACAAGTTTGTACAAAAAAGCAGGCTCCGAATTCG
CCCTT

3' Linker Sequence: None

 

 Recommended Growth Condition:

Distributed in bacterial strain : DH5-alpha T1 phage resistant
Antibiotic Selection: Host Type:  bacterial    Marker:  ampicillin
Host Type:  bacterial    Marker:  chloramphenicol
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: None
Recommended expression in: Not Applicable
       DyNA Vector Map

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Vector Name: pVP16

pVP16 in advanced viewed

Synonyms: None
Sequencing Primer: Forward:  MBP Forward
Reverse:  pQE Reverse
Description: Bacterial (E. coli) expression, adds N-terminal 8xHis and MBP, adds TEV site on 5p end of insert; ampicillin resistance in bacteria; recombinational cloning; Gateway cloning into this vector will remove the Gateway ccdB death casette.
Comments: TEV protease cleavage site is "partial" and not functional. The TEV cleavage site must be added to the insert by PCR, which allows amino acids encoded by the attB1 site to be cut off after protein expression. Gateway cloning into this vector will remove the Gateway cassette
Size (bp): 7631
Parent Vector: None
Empty Vector: EvNO00084279
Properties: Gateway, acceptor (destination), bacterial expression, recombinational cloning, with tag/fusion/marker
Author Name: Center for Eukaryotic Structural Genomics
Publications: PMID: 15750721
Title: Results from high-throughput DNA cloning of Arabidopsis thaliana target genes using site-specific recombination

Vector Map:         Vector Sequence:
Sequence:

 



Vector Features:

Type Name Description Start Position End Position
bacterial origin ori ColE1-type bacterial origin of replication 6471 5789
negative selection marker ccdB ccdB death cassette gene (lost in with-insert form) 2643 2852
promoter T5 T5 promoter 5 54
recombination site AttR1 AttR1 recombination site 1316 1440
recombination site AttR2 AttR2 recombination site 3020 2896
repressor protein gene LacI LacI coding region 4132 5211
selectable marker AmpR ampicillin resistance gene 7228 6569
selectable marker CmR chloramphenicol resistance gene (CmR) (Lost in with-insert form) 1549 2205
tag His N-terminal 8xHis tag 128 145
tag MBP N-terminal MBP 148 1246

 

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Cloning Information : 724504
Original Clone ID : MBP-linker-MPR-TM