pDual-eGFP(H66)

63558 Synthetic Bacterial Expression

Plasmid Information

Purpose
Expression of the Azure (blue) spectral variant of eGFP in bacteria and in mammalian cells. Could be used as a reporter for quantifying gene targeting and recombineering in mammalian cells.
Depositor
Richard S Myers
Article
Valledor et al IUBMB Life. 2012 Aug;64(8):684-9. doi: 1
Insert
His-T7-eGFP(H66)
Tags
His6 and T7
Mutation
Changed tyrosine at position 66 with respect to t…
Promoter
CMV-EF1α hybrid (CEF)
Available Since
April 15, 2015
Availability
Available

Sequence Information

Length
825 bp
Topology
linear
Molecule Type
genomic DNA
View on AddGene.org

Features

CDS 4
Label Gene Product Position Strand
6xHis — 6xHis affinity tag 167..185 Fwd
DNA Sequence
CATCATCATCATCATCAC
Protein Sequence
HHHHHH
thrombin site — thrombin recognition and cleavage site 194..212 Fwd
DNA Sequence
CTGGTGCCGCGCGGCAGC
Protein Sequence
LVPRGS
T7 tag (gene 10 leader) — leader peptide from bacteriophage T7 ge… 215..248 Fwd
DNA Sequence
ATGGCTAGCATGACTGGTGGACAGCAAATGGGT
Protein Sequence
MASMTGGQQMG
VN173 — N-terminal fragment of mVenus for use i… 272..791 Fwd
DNA Sequence
ATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCACCCTGACCCACGGCGTGCAGTGCTTCAGCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCACAACGTCTATATCATGGCCGACAAGCAGAAGAACGGCATCAAGGTGAACTTCAAGATCCGCCACAACATCGAG
Protein Sequence
MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIE
promoter 1
Label Gene Product Position Strand
T7 promoter — — 68..87 Fwd
DNA Sequence
TAATACGACTCACTATAGG
protein_bind 1
Label Gene Product Position Strand
lac operator — — 87..112 Fwd
DNA Sequence
GGAATTGTGAGCGGATAACAATTCC