pET GFP plasmid is a void vector. Your quality can be embedded with a LIC cloning convention.

This vector includes a short GFP space 11 tag to the C-end of your protein. After expression and sanitization, your protein can be blended with a "biosensor" atom (GFP spaces 1-10), and fluorescence will create. This strategy can be helpful if straightforwardly intertwining the whole GFP particle meddles with the capacity of your protein.

To clone into this vector, add LIC labels to the 5' end of your PCR preliminaries.

Forward - 5' TACTTCCAATCCAATGCA3'

Turn around - 5' CTCCCACTACCAATGCC 3'

Try not to incorporate a stop codon with your turn around preliminary.

Linearize the plasmid with SspI, then gel filter.

While processing the DNA with T4 polymerase, utilize dCTP for the embed and dGTP for your linearized vector.

pET GFP domain 11 LIC cloning vector (GFP C-11)

Item
Catalog #
Description
Quantity
Price (USD)

Plasmid
30183
Plasmid sent as bacteria in agar stab
1
$65


Backbone
Vector backbone
pET
Backbone size (bp) 5422
Vector type
Bacterial Expression


Growth in Bacteria

Bacterial Resistance(s)
Kanamycin
Growth Temperature
37°C
Growth Strain(s)
DH5alpha
Copy number
Low Copy

Gene/Insert

Gene/Insert name
None
Tags / Fusion Proteins
GFP domain 11 (C terminal on backbone)
Biotin-H6-TEV recognition (N terminal on backbone)

Cloning Information

Cloning method Restriction Enzyme
5′ cloning site LIC site vGFP (destroyed during cloning)
3′ cloning site LIC site vGFP (destroyed during cloning)
5′ sequencing primer T7 forward
3′ sequencing primer T7 reverse