PA-DAD-dark

53096 Mus musculus Bacterial Expression

Plasmid Information

Purpose
Expresses dark mutant PA-DAD in mammalian cells, an optogenetic tool to activate endogenous diaphanous related formins in live cells using light with LOV2 in the closed conformation
Depositor
Klaus Hahn, Ronen Zaidel-Bar
Article
Rao et al Cytoskeleton (Hoboken). 2013 Jul;70(7):3
Insert
LOV(dark)-6aa-DAD
Tags
6x His and mVenus
Mutation
L47F and C49S in Venus; C450M mutation in LOV2
Available Since
Aug. 19, 2015
Availability
Available

Sequence Information

Length
1,112 bp
Topology
linear
Molecule Type
genomic DNA
View on AddGene.org

Features

CDS 2
Label Gene Product Position Strand
6xHis — 6xHis affinity tag 208..226 Fwd
DNA Sequence
CACCATCACCACCATCAC
Protein Sequence
HHHHHH
mGold — photostable derivative of mVenus (Lee e… 232..946 Fwd
DNA Sequence
GTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCCTGAAGTTCATCAGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCACCCTCGGCTACGGCCTGCAGTGCTTCGCCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCACAACGTCTATATCACCGCCGACAAGCAGAAGAACGGCATCAAGGCAAACTTCAAGATCCGCCACAACATCGAGGACGGCGGCGTGCAGCTCGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCTACCAGTCCAAACTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCTGTACAAG
Protein Sequence
VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFISTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK
promoter 2
Label Gene Product Position Strand
T7 promoter — — 19..38 Fwd
DNA Sequence
TAATACGACTCACTATAGG
p10 promoter — — 78..188 Fwd
DNA Sequence
GACCTTTAATTCAACCCAACACAATATATTATAGTTAAATAAGAATTATTATCAAATCATTTGTATATTAATTAAAATACTATACTGTAAATTACATTTTATTTACAATC
protein_bind 1
Label Gene Product Position Strand
lac operator — — 38..63 Fwd
DNA Sequence
GGAATTGTGAGCGGATAACAATTCC