CL7_2xNLS_iGeoCas9(C1)_2xNLS_His

222987 Synthetic Bacterial Expression

Plasmid Information

Purpose
Bacterial expression of iGeoCas9(C1) construct
Depositor
Jennifer Doudna
Article
Chen et al bioRxiv [Preprint]. 2023 Nov 15:2023.11.
Insert
CL7_2xNLS_iGeoCas9(C1)_2xNLS_His
Tags
CL7 and His
Promoter
tac
Available Since
July 26, 2024
Availability
Available

Sequence Information

Length
7,657 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

3'UTR 1
Label Gene Product Position Strand
cspA 3'UTR cspA — 4052..4197 Fwd
DNA Sequence
TGCGCATTCTCATTGCACCCAAATTTATTCTTCACAAAAATAATAATAGATTTTATTACGCGATCGATTATTTATTTCCTGAAAACAAATAAAAAAATCCCCGCCAAATGGCAGGGATCTTAGATTCTGTGCTTTTAAGCAGAGA
5'UTR 1
Label Gene Product Position Strand
cspA 5'UTR cspA — 438..572 Fwd
DNA Sequence
GGTGTATTACCTCTTAATAATTAAGTGTGCCTTTCGGCGATATGGCGTGCTTTACAGATTTTGAAGCGTTAAAGGAATGTGCACTACGAGGGGTATCAACGATAACTCTTGAAGGGACTTGCCTTACTACACTG
CDS 9
Label Gene Product Position Strand
HRV 3C site — recognition and cleavage site for human… 6..30 Rev
DNA Sequence
TTGGAGGTTTTGTTCCAGGGTCCA
Protein Sequence
LEVLFQGP
lacI lacI lac repressor 821..1904 Fwd
DNA Sequence
GTGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGA
Protein Sequence
MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ
AmpR bla beta-lactamase 2834..3695 Rev
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
6xHis — 6xHis affinity tag 4207..4225 Rev
DNA Sequence
CACCATCACCATCACCAT
Protein Sequence
HHHHHH
HRV 3C site — recognition and cleavage site for human… 4231..4255 Rev
DNA Sequence
TTAGAAGTGCTTTTTCAGGGCCCA
Protein Sequence
LEVLFQGP
SV40 NLS — nuclear localization signal of SV40 (si… 4258..4279 Rev
DNA Sequence
CCTAAGAAGAAGAGAAAAGTT
Protein Sequence
PKKKRKV
SV40 NLS — nuclear localization signal of SV40 (si… 4288..4309 Rev
DNA Sequence
CCTAAAAAAAAGAGAAAAGTA
Protein Sequence
PKKKRKV
SV40 NLS — nuclear localization signal of SV40 (si… 7603..7624 Rev
DNA Sequence
CCGAAAAAGAAACGGAAAGTA
Protein Sequence
PKKKRKV
SV40 NLS — nuclear localization signal of SV40 (si… 7633..7654 Rev
DNA Sequence
CCAAAGAAAAAGCGTAAAGTC
Protein Sequence
PKKKRKV
promoter 3
Label Gene Product Position Strand
tac promoter — — 615..644 Rev
DNA Sequence
TTGACAATTAATCATCGGCTCGTATAATG
lacI promoter lacI — 743..821 Fwd
DNA Sequence
GACACCATCGAATGGCGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAAT
AmpR promoter bla — 3695..3800 Rev
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
protein_bind 1
Label Gene Product Position Strand
lac operator — — 593..610 Fwd
DNA Sequence
TTGTTATCCGCTCACAA
rep_origin 1
Label Gene Product Position Strand
ori — — 2075..2664 Rev
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 1
Label Gene Product Position Strand
rrnB T1 terminator Escherichia coli rrnB — 3905..3977 Rev
DNA Sequence
CAAATAAAACGAAAGGCTCAGTCGAAAGACTGGGCCTTTCGTTTTATCTGTTGTTTGTCGGTGAACGCTCTC