pHAGE UbiC NLS-HA-stdMCP-stdHalo-RH1

164614 Synthetic Bacterial Expression

Plasmid Information

Purpose
Lentiviral vector expressing synonymous tandem MS2 coat protein fused to synonymous tandem Halo-tag and the RH1 domain of RILPL2. Used for single-molecule imaging of actin-anchored mRNA.
Depositor
Jeffrey Chao
Article
Mateju et al Cell. 2020 Dec 23;183(7):1801-1812.e13.
Insert
stdMCP-stdHalo-RH1
Tags
HA tag and NLS
Promoter
UbiC
Available Since
Feb. 18, 2021
Availability
Available

Sequence Information

Length
8,317 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 8
Label Gene Product Position Strand
gp41 peptide — antigenic peptide corresponding to amin… 1720..1765 Fwd
DNA Sequence
AAGAATGAACAAGAATTATTGGAATTAGATAAATGGGCAAGTTTG
Protein Sequence
KNEQELLELDKWASL
SV40 NLS — nuclear localization signal of SV40 (si… 2635..2656 Fwd
DNA Sequence
CCAAAAAAGAAAAGAAAAGTT
Protein Sequence
PKKKRKV
HA — HA (human influenza hemagglutinin) epit… 2659..2686 Fwd
DNA Sequence
TACCCCTACGACGTGCCCGACTACGCC
Protein Sequence
YPYDVPDYA
HRV 3C site — recognition and cleavage site for human… 2686..2710 Fwd
DNA Sequence
CTGGAAGTTCTGTTCCAGGGGCCC
Protein Sequence
LEVLFQGP
HaloTag(R) — modified bacterial dehalogenase that fo… 3448..4336 Fwd
DNA Sequence
ATGGAGATCGGCACCGGCTTCCCCTTTGACCCTCACTACGTGGAGGTGCTGGGCGAGAGGATGCACTATGTGGACGTGGGACCAAGAGATGGAACCCCCGTGCTGTTCCTGCACGGCAACCCCACAAGCTCCTACGTGTGGCGGAATATCATCCCACACGTGGCCCCCACACACAGATGCATCGCCCCAGATCTGATCGGCATGGGCAAGTCCGACAAGCCCGATCTGGGCTATTTCTTTGACGATCACGTGCGGTTCATGGACGCCTTTATCGAGGCCCTGGGCCTGGAGGAGGTGGTGCTGGTCATCCACGATTGGGGCTCTGCCCTGGGCTTTCACTGGGCCAAGCGGAACCCCGAGAGAGTGAAGGGCATCGCCTTCATGGAGTTTATCAGACCAATCCCAACCTGGGATGAGTGGCCTGAGTTCGCCAGAGAGACATTCCAGGCCTTTCGGACCACAGACGTGGGCAGAAAGCTGATCATCGATCAGAACGTGTTTATCGAAGGCACCCTGCCTATGGGAGTGGTGAGGCCACTGACAGAGGTGGAGATGGACCACTACCGCGAGCCTTTCCTGAATCCAGTGGATAGGGAGCCTCTGTGGCGCTTTCCCAACGAGCTGCCTATCGCCGGCGAGCCAGCCAATATCGTGGCCCTGGTGGAGGAGTATATGGACTGGCTGCACCAGTCCCCTGTGCCAAAGCTGCTGTTCTGGGGCACCCCAGGCGTGCTGATCCCTCCCGCAGAGGCAGCAAGGCTGGCCAAGTCTCTGCCCAATTGTAAGGCCGTGGATATCGGCCCTGGCCTGAACCTGCTGCAGGAGGACAATCCTGATCTGATCGGCAGCGAGATCGCCCGCTGGCTGTCTACACTGGAGATCAGCGGC
Protein Sequence
MEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIGSEIARWLSTLEISG
HRV 3C site — recognition and cleavage site for human… 4348..4372 Fwd
DNA Sequence
CTGGAAGTTCTGTTTCAGGGCCCG
Protein Sequence
LEVLFQGP
Factor Xa site — Factor Xa recognition and cleavage site 5177..5189 Rev
DNA Sequence
ATTGAGGGCCGA
Protein Sequence
IEGR
AmpR bla beta-lactamase 5912..6773 Fwd
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
LTR 2
Label Gene Product Position Strand
3' LTR — — 0..634 Fwd
DNA Sequence
TGGAAGGGCTAATTCACTCCCAAAGAAGACAAGATATCCTTGATCTGTGGATCTACCACACACAAGGCTACTTCCCTGATTAGCAGAACTACACACCAGGGCCAGGGGTCAGATATCCACTGACCTTTGGATGGTGCTACAAGCTAGTACCAGTTGAGCCAGATAAGGTAGAAGAGGCCAATAAAGGAGAGAACACCAGCTTGTTACACCCTGTGAGCCTGCATGGGATGGATGACCCGGAGAGAGAAGTGTTAGAGTGGAGGTTTGACAGCCGCCTAGCATTTCATCACGTGGCCCGAGAGCTGCATCCGGAGTACTTCAAGAACTGCTGATATCGAGCTTGCTACAAGGGACTTTCCGCTGGGGACTTTCCAGGGAGGCGTGGCCTGGGCGGGACTGGGGAGTGGCGAGCCCTCAGATCCTGCATATAAGCAGCTGCTTTTTGCCTGTACTGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTGGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGTAGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGTCAGTGTGGAAAATCTCTAGCA
3' LTR (Delta-U3) — — 5369..5603 Fwd
DNA Sequence
TGGAAGGGCTAATTCACTCCCAACGAAGACAAGATCTGCTTTTTGCTTGTACTGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTGGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGTAGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGTCAGTGTGGAAAATCTCTAGCA
misc_feature 3
Label Gene Product Position Strand
RRE — — 1302..1536 Fwd
DNA Sequence
AGGAGCTTTGTTCCTTGGGTTCTTGGGAGCAGCAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGACGGTACAGGCCAGACAATTATTGTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTGAGGCGCAACAGCATCTGTTGCAACTCACAGTCTGGGGCATCAAGCAGCTCCAGGCAAGAATCCTGGCTGTGGAAAGATACCTAAAGGATCAACAGCTCCT
cPPT/CTS — — 2057..2175 Fwd
DNA Sequence
TTTTAAAAGAAAAGGGGGGATTGGGGGGTACAGTGCAGGGGAAAGAATAGTAGACATAATAGCAACAGACATACAAACTAAAGAATTACAAAAACAAATTACAAAAATTCAAAATTTT
WPRE — — 4706..5295 Fwd
DNA Sequence
AATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGC
promoter 3
Label Gene Product Position Strand
UbC promoter — — 2217..2616 Fwd
DNA Sequence
GGCCTCCGCGCCGGGTTTTGGCGCCTCCCGCGGGCGCCCCCCTCCTCACGGCGAGCGCTGCCACGTCAGACGAAGGGCGCAGCGAGCGTCCTGATCCTTCCGCCCGGACGCTCAGGACAGCGGCCCGCTGCTCATAAGACTCGGCCTTAGAACCCCAGTATCAGCAGAAGGACATTTTAGGACGGGACTTGGGTGACTCTAGGGCACTGGTTTTCTTTCCAGAGAGCGGAACAGGCGAGGAAAAGTAGTCCCTTCTCGGCGATTCTGTGGAGGGATCTCCGTGGGGCGGTGAACGCCGATGATTATATAAGGACGCGCCGGGTGTGGCACAGCTAGTTCCGTCGCAGCCGGGATTTGGGTCGCGGTTCTTGTTTGTGGATCGCTGTGATCGTCACTTGG
AmpR promoter bla — 5807..5912 Fwd
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
lac promoter — — 7978..8009 Fwd
DNA Sequence
TTTACACTTTATGCTTCCGGCTCGTATGTTG
protein_bind 2
Label Gene Product Position Strand
CAP binding site — — 7942..7964 Fwd
DNA Sequence
TAATGTGAGTTAGCTCACTCAT
lac operator — — 8016..8033 Fwd
DNA Sequence
TTGTGAGCGGATAACAA
rep_origin 1
Label Gene Product Position Strand
ori — — 6943..7532 Fwd
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA