Addgene - pFastBac Dual His6 MBP N10 TEV LIC cloning vector (5C) Plasmid Data

Plasmid 30123: pFastBac Dual His6 MBP N10 TEV LIC cloning vector (5C)

Gene/insert name:
None
Fusion protein or tag:
His6-MBP-N10-TEV
Terminal:
N terminal on backbone
Vector backbone:
pFastBac Dual
(Search Vector Database)
Vector type:
Insect Expression
Backbone size (bp):
6472
Cloning site 5':
LIC site
Site destroyed during cloning:
Yes
Cloning site 3':
LIC site
Site destroyed during cloning:
Yes
5' sequencing primer:
see notes List of Sequencing Primers
3' sequencing primer:
see notes
Bacterial resistance(s)
Ampicillin
Growth strain(s)
DH5alpha
Growth temperature (℃):
37
High or low copy:
High Copy
Selectable markers:
Gentamicin
Sequence: View sequences (3)
Principal Investigator:
Scott Gradia
Terms and Licenses:
MTA

Comments:

This plasmid is a LIC-adapted pFastBac Dual vector. It uses the same PCR primer tags as with most of our other vectors, so one PCR product can be inserted into many different vectors at once.

This vector is a dual expression vector, and your genes can be inserted in any order (check your gene for internal restriction sites, as this may dictate cloning order).

A gene inserted into LIC site 1 will have a His6-MBP-N10-TEV tag added to the N-terminus. A gene inserted into LIC site 2 will remain untagged.

LIC site v1:
Add the following tags to your PCR primers:

LicV1 Forward Tag TACTTCCAATCCAATGCA

LicV1 Reverse Tag TTATCCACTTCCAATGTTATTA

Linearize vector with SspI and gel purify. T4-treat vector with dGTP. For the PCR product, T4-treat with dCTP.

LIC site v2:
Add the following tags to your PCR primers:

LicV2 Forward Tag TTTAAGAAGGAGATATAGATC(ATG)

LicV2 Reverse Tag TTATGGAGTTGGGATCTTATTA

Linearize vector with EcoRV and gel purify. T4-treat vector with dCTP. For the PCR product, T4-treat with dGTP.

For more information, please see our website:
http://qb3.berkeley.edu/qb3/macrolab/

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 30123" in your Materials and Methods section.