pMT-Hrp48-ADARcd-E488Q-V5

Plasmid Information

Purpose

Expresses the hyperactive form of the catalytic domain of the ADAR protein fused to HRP-48 for use in "TRIBE" assays

Depositor

Michael Rosbash

Article

Xu et al RNA. 2018 Feb;24(2):173-182. doi: 10.126

Insert

Hrp48, ADAR (Adar Fly)

Sequence Information

Length

7,362 bp

Topology

circular

Molecule Type

other DNA

View on AddGene.org

Features

CDS 4

Label	Gene	Product	Position	Strand
V5 tag	—	epitope tag from simian virus 5	3519..3561	Fwd
DNA Sequence GGTAAGCCTATCCCTAACCCTCTCCTCGGTCTCGATTCTACG Protein Sequence GKPIPNPLLGLDST
6xHis	—	6xHis affinity tag	3570..3588	Fwd
DNA Sequence CATCATCACCATCACCAT Protein Sequence HHHHHH
BSD	Aspergillus terreus BSD	blasticidin S deaminase	4502..4901	Fwd
DNA Sequence ATGGCCAAGCCTTTGTCTCAAGAAGAATCCACCCTCATTGAAAGAGCAACGGCTACAATCAACAGCATCCCCATCTCTGAAGACTACAGCGTCGCCAGCGCAGCTCTCTCTAGCGACGGCCGCATCTTCACTGGTGTCAATGTATATCATTTTACTGGGGGACCTTGCGCAGAACTCGTGGTGCTGGGCACTGCTGCTGCTGCGGCAGCTGGCAACCTGACTTGTATCGTCGCGATCGGAAATGAGAACAGGGGCATCTTGAGCCCCTGCGGACGGTGCCGACAGGTTCTTCTCGATCTGCATCCTGGGATCAAAGCCATAGTGAAGGACAGTGATGGACAGCCGACGGCAGTTGGGATTCGTGAATTGCTGCCCTCTGGTTATGTGTGGGAGGGCTAA Protein Sequence MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGVNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIKAIVKDSDGQPTAVGIRELLPSGYVWEG
AmpR	bla	beta-lactamase	6301..7162	Rev
DNA Sequence ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA Protein Sequence MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW

polyA_signal 2

Label	Gene	Product	Position	Strand
SV40 poly(A) signal	—	—	3629..3764	Fwd
DNA Sequence GATCCAGACATGATAAGATACATTGATGAGTTTGGACAAACCACAACTAGAATGCAGTGAAAAAAATGCTTTATTTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATAAACAAGTT
SV40 poly(A) signal	—	—	5229..5364	Fwd
DNA Sequence AACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGATC

promoter 4

Label	Gene	Product	Position	Strand
MT promoter	—	—	400..827	Fwd
DNA Sequence AATTCGTTGCAGGACAGGATGTGGTGCCCGATGTGACTAGCTCTTTGCTGCAGGCCGTCCTATCCTCTGGTTCCGATAAGAGACCCAGAACTCCGGCCCCCCACCGCCCACCGCCACCCCCATACATATGTGGTACGCAAGTAAGAGTGCCTGCGCATGCCCCATGTGCCCCACCAAGAGCTTTGCATCCCATACAAGTCCCCAAAGTGGAGAACCGAACCAATTCTTCGCGGGCAGAACAAAAGCTTCTGCACACGTCTCCACTCGAATTTGGAGCCGGCCGGCGTGTGCAAAAGAGGTGAATCGAACGAAAGACCCGTGTGTAAAGCCGCGTTTCCAAAATGTATAAAACCGAGAGCATCTGGCCAATGTGCATCAGTTGTGGTCAGCAGCAAAATCAAGTGAATCATCTCAGTGCAACTAAAGG
lac promoter	—	—	3955..3986	Rev
DNA Sequence TTTACACTTTATGCTTCCGGCTCGTATGTTG
copia promoter	—	—	4206..4486	Fwd
DNA Sequence TGTTGGAATATACTATTCAACCTACAAAAGTAACGTTAAACAACACTACTTTATATTTGATATGAATGGCCACACCTTTTATGCCATAAAACATATTGTAAGAGAATACCACTCTTTTTATTCCTTCTTTCCTTCTTGTACGTTTTTTGCTGTAAGTAGGTCGTGGTGCTGGTGTTGCAGTTGAAATAACTTAAAATATAAATCATAAAACTCAAACATAAACTTGACTATTTATTTATTTATTAAGAAAGGAAATATAAATTGTAAATTACAACAGGTT
AmpR promoter	bla	—	7162..7267	Rev
DNA Sequence CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT

protein_bind 2

Label	Gene	Product	Position	Strand
lac operator	—	—	3931..3948	Fwd
DNA Sequence TTGTTATCCGCTCACAA
CAP binding site	—	—	4000..4022	Fwd
DNA Sequence ATGAGTGAGCTAACTCACATTA

rep_origin 1

Label	Gene	Product	Position	Strand
ori	—	—	5542..6131	Rev
DNA Sequence TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA