pCDF-5xT7-TtCsm

128572 Synthetic Bacterial Expression

Plasmid Information

Purpose
For expression of T. thermophilus Csm in E. coli (Csm1-5)
Depositor
Jennifer Doudna
Article
Liu et al Nat Commun. 2019 Jul 5;10(1):3001. doi:
Tags
HRV 3C protease cleavage site and 10xHis tag on C…
Mutation
Codon optimized for expression in E. coli and Cod…
Promoter
T7 promoter, lac operator
Available Since
Aug. 6, 2019
Availability
Available

Sequence Information

Length
9,479 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 4
Label Gene Product Position Strand
HRV 3C site — recognition and cleavage site for human… 6073..6097 Fwd
DNA Sequence
CTGGAAGTTCTGTTCCAGGGGCCG
Protein Sequence
LEVLFQGP
10xHis — 10xHis affinity tag 6097..6127 Fwd
DNA Sequence
CATCATCACCACCATCACCATCATCATCAC
Protein Sequence
HHHHHHHHHH
SmR aadA aminoglycoside adenylyltransferase (Mur… 6377..7169 Rev
DNA Sequence
ATGAGGGAAGCGGTGATCGCCGAAGTATCGACTCAACTATCAGAGGTAGTTGGCGTCATCGAGCGCCATCTCGAACCGACGTTGCTGGCCGTACATTTGTACGGCTCCGCAGTGGATGGCGGCCTGAAGCCACACAGTGATATTGATTTGCTGGTTACGGTGACCGTAAGGCTTGATGAAACAACGCGGCGAGCTTTGATCAACGACCTTTTGGAAACTTCGGCTTCCCCTGGAGAGAGCGAGATTCTCCGCGCTGTAGAAGTCACCATTGTTGTGCACGACGACATCATTCCGTGGCGTTATCCAGCTAAGCGCGAACTGCAATTTGGAGAATGGCAGCGCAATGACATTCTTGCAGGTATCTTCGAGCCAGCCACGATCGACATTGATCTGGCTATCTTGCTGACAAAAGCAAGAGAACATAGCGTTGCCTTGGTAGGTCCAGCGGCGGAGGAACTCTTTGATCCGGTTCCTGAACAGGATCTATTTGAGGCGCTAAATGAAACCTTAACGCTATGGAACTCGCCGCCCGACTGGGCTGGCGATGAGCGAAATGTAGTGCTTACGTTGTCCCGCATTTGGTACAGCGCAGTAACCGGCAAAATCGCGCCGAAGGATGTCGCTGCCGACTGGGCAATGGAGCGCCTGCCGGCCCAGTATCAGCCCGTCATACTTGAAGCTAGACAGGCTTATCTTGGACAAGAAGAAGATCGCTTGGCCTCGCGCGCAGATCAGTTGGAAGAATTTGTCCACTACGTGAAAGGCGAGATCACCAAGGTAGTCGGCAAATAA
Protein Sequence
MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK
lacI lacI lac repressor 8256..9339 Rev
DNA Sequence
GTGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGACATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGA
Protein Sequence
MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ
promoter 7
Label Gene Product Position Strand
T7 promoter — — 0..9479 Fwd
DNA Sequence
TAATACGACTCACTATAGG
T7 promoter — — 2503..2522 Fwd
DNA Sequence
TAATACGACTCACTATAGG
T7 promoter — — 3001..3020 Fwd
DNA Sequence
TAATACGACTCACTATAGG
T7 promoter — — 3820..3839 Fwd
DNA Sequence
TAATACGACTCACTATAGG
T7 promoter — — 4789..4808 Fwd
DNA Sequence
TAATACGACTCACTATAGG
AmpR promoter bla — 7169..7261 Rev
DNA Sequence
TTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
lacI promoter lacI — 9339..9417 Rev
DNA Sequence
GACACCATCGAATGGCGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAAT
protein_bind 5
Label Gene Product Position Strand
lac operator — — 2..27 Fwd
DNA Sequence
GGAATTGTGAGCGGATAACAATTCC
lac operator — — 2522..2547 Fwd
DNA Sequence
GGAATTGTGAGCGGATAACAATTCC
lac operator — — 3020..3045 Fwd
DNA Sequence
GGAATTGTGAGCGGATAACAATTCC
lac operator — — 3839..3864 Fwd
DNA Sequence
GGAATTGTGAGCGGATAACAATTCC
lac operator — — 4808..4833 Fwd
DNA Sequence
GGAATTGTGAGCGGATAACAATTCC
rep_origin 1
Label Gene Product Position Strand
CloDF13 ori — — 7308..8047 Rev
DNA Sequence
GATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTTTTGCCCTGTAAACGAAAAAACCACCTGGGGAGGTGGTTTGATCGAAGGTTAAGTCAGTTGGGGAACTGCTTAACCGTGGTAACTGGCTTTCGCAGAGCACAGCAACCAAATCTGTCCTTCCAGTGTAGCCGGACTTTGGCGCACACTTCAAGAGCAACCGCGTGTTTAGCTAAACAAATCCTCTGCGAACTCCCAGTTACCAATGGCTGCTGCCAGTGGCGTTTTACCGTGCTTTTCCGGGTTGGACTCAAGTGAACAGTTACCGGATAAGGCGCAGCAGTCGGGCTGAACGGGGAGTTCTTGCTTACAGCCCAGCTTGGAGCGAACGACCTACACCGAGCCGAGATACCAGTGTGTGAGCTATGAGAAAGCGCCACACTTCCCGTAAGGGAGAAAGGCGGAACAGGTATCCGGTAAACGGCAGGGTCGGAACAGGAGAGCGCAAGAGGGAGCGACCCGCCGGAAACGGTGGGGATCTTTAAGTCCTGTCGGGTTTCGCCCGTACTGTCAGATTCATGGTTGAGCCTCACGGCTCCCACAGATGCACCGGAAAAGCGTCTGTTTATGTGAACTCTGGCAGGAGGGCGGAGCCTATGGAAAAACGCCACCGGCGCGGCCCTGCTGTTTTGCCTCACATGTTAGTCCCCTGCTTATCCACGGAATCTGTGGGTAACTTTGTATGTGTCCGCAGCGC
terminator 1
Label Gene Product Position Strand
T7 terminator — — 6159..6207 Fwd
DNA Sequence
CTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG