LbCpf1-2NLS

102566 Synthetic Bacterial Expression

Plasmid Information

Purpose
bacterial expression of LbCpf1-2NLS
Depositor
Jennifer Doudna
Article
Moreno-Mateos et al Nat Commun. 2017 Dec 8;8(1):2024. doi: 1
Insert
LbCpf1
Tags
6xHis-MBP-TEV and HA-2xSV40NLS
Promoter
T7
Available Since
Dec. 21, 2017
Availability
Available

Sequence Information

Length
10,514 bp
Topology
circular
Molecule Type
other DNA

Features

CDS 12
Label Gene Product Position Strand
6xHis 6xHis affinity tag 757..775 Fwd
DNA Sequence
CACCATCACCATCACCAT
Protein Sequence
HHHHHH
MBP malE (mutated) maltose binding protein from E. coli 784..1885 Fwd
DNA Sequence
ATGAAAATCGAAGAAGGTAAACTGGTAATCTGGATTAACGGCGATAAAGGCTATAACGGTCTCGCTGAAGTCGGTAAGAAATTCGAGAAAGATACCGGAATTAAAGTCACCGTTGAGCATCCGGATAAACTGGAAGAGAAATTCCCACAGGTTGCGGCAACTGGCGATGGCCCTGACATTATCTTCTGGGCACACGACCGCTTTGGTGGCTACGCTCAATCTGGCCTGTTGGCTGAAATCACCCCGGACAAAGCGTTCCAGGACAAGCTGTATCCGTTTACCTGGGATGCCGTACGTTACAACGGCAAGCTGATTGCTTACCCGATCGCTGTTGAAGCGTTATCGCTGATTTATAACAAAGATCTGCTGCCGAACCCGCCAAAAACCTGGGAAGAGATCCCGGCGCTGGATAAAGAACTGAAAGCGAAAGGTAAGAGCGCGCTGATGTTCAACCTGCAAGAACCGTACTTCACCTGGCCGCTGATTGCTGCTGACGGGGGTTATGCGTTCAAGTATGAAAACGGCAAGTACGACATTAAAGACGTGGGCGTGGATAACGCTGGCGCGAAAGCGGGTCTGACCTTCCTGGTTGACCTGATTAAAAACAAACACATGAATGCAGACACCGATTACTCCATCGCAGAAGCTGCCTTTAATAAAGGCGAAACAGCGATGACCATCAACGGCCCGTGGGCATGGTCCAACATCGACACCAGCAAAGTGAATTATGGTGTAACGGTACTGCCGACCTTCAAGGGTCAACCATCCAAACCGTTCGTTGGCGTGCTGAGCGCAGGTATTAACGCCGCCAGTCCGAACAAAGAGCTGGCAAAAGAGTTCCTCGAAAACTATCTGCTGACTGATGAAGGTCTGGAAGCGGTTAATAAAGACAAACCGCTGGGTGCCGTAGCGCTGAAGTCTTACGAGGAAGAGTTGGCGAAAGATCCACGTATTGCCGCCACTATGGAAAACGCCCAGAAAGGTGAAATCATGCCGAACATCCCGCAGATGTCCGCTTTCTGGTATGCCGTGCGTACTGCGGTGATCAACGCCGCCAGCGGTCGTCAGACTGTCGATGAAGCCCTGAAAGACGCGCAGACT
Protein Sequence
MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT
TEV site tobacco etch virus (TEV) protease recog… 1939..1960 Fwd
DNA Sequence
GAAAACCTGTACTTCCAATCC
Protein Sequence
ENLYFQS
LbCpf1 CRISPR-associated protein from Lachnosp… 1969..5653 Fwd
DNA Sequence
ATGAGCAAACTGGAGAAGTTCACGAACTGTTATTCTCTGTCGAAAACCTTACGTTTTAAGGCCATTCCGGTAGGTAAAACGCAAGAAAATATCGACAATAAACGTCTCCTGGTTGAAGATGAAAAGCGTGCCGAAGATTATAAAGGTGTTAAAAAGCTGCTTGATCGCTATTACCTGTCTTTTATCAACGATGTTCTGCATTCCATCAAATTGAAAAACTTGAATAATTACATCTCGTTATTTCGCAAAAAAACCCGTACCGAAAAAGAAAACAAGGAGCTGGAAAACCTTGAAATTAATCTGCGTAAAGAAATCGCGAAAGCATTTAAAGGTAATGAAGGATATAAATCTCTGTTTAAAAAAGACATTATTGAAACCATCCTGCCTGAGTTCTTGGATGATAAAGATGAAATTGCCCTGGTTAATAGCTTTAATGGCTTTACGACTGCATTTACCGGCTTTTTTGATAACCGTGAAAACATGTTCAGCGAAGAAGCGAAATCGACTTCCATTGCCTTCCGTTGCATTAATGAAAACCTGACCCGCTACATTTCTAATATGGACATCTTTGAAAAAGTTGATGCCATTTTCGACAAACATGAAGTTCAAGAAATCAAAGAAAAGATTTTAAACTCCGATTATGATGTTGAAGATTTCTTTGAGGGAGAATTTTTCAATTTTGTTCTCACCCAAGAGGGTATTGATGTCTATAACGCCATTATCGGAGGATTTGTTACGGAATCAGGCGAAAAGATTAAAGGACTGAATGAATACATTAATTTATACAATCAGAAAACCAAACAGAAATTGCCAAAATTCAAACCACTGTATAAACAGGTATTAAGCGACCGTGAATCCTTGTCCTTTTATGGCGAAGGCTATACAAGCGATGAAGAGGTGCTCGAAGTATTCCGTAATACTTTGAACAAAAACTCGGAAATCTTCTCCAGCATTAAAAAACTGGAAAAATTGTTTAAAAATTTCGATGAATATAGTAGCGCAGGCATCTTCGTCAAAAATGGCCCTGCGATTTCAACCATTTCTAAAGACATTTTCGGTGAATGGAACGTGATCCGCGATAAATGGAACGCGGAGTATGACGACATTCATCTGAAAAAAAAAGCCGTTGTTACCGAAAAGTATGAAGATGACCGCCGCAAGAGCTTTAAAAAAATTGGTAGTTTCTCACTGGAACAGTTACAAGAATATGCAGACGCCGATCTGTCGGTTGTGGAAAAGTTAAAAGAGATTATTATCCAGAAAGTCGATGAAATTTATAAAGTGTACGGCTCCAGCGAAAAGCTCTTTGATGCCGACTTTGTTTTAGAAAAATCGCTGAAAAAAAACGATGCCGTGGTGGCCATCATGAAAGATCTGTTGGATTCGGTCAAATCGTTCGAAAACTATATCAAGGCGTTCTTTGGCGAAGGGAAAGAGACCAATCGTGACGAATCGTTCTATGGCGATTTTGTGTTGGCGTATGATATTCTTCTGAAAGTAGATCACATCTATGATGCTATCCGCAACTACGTGACCCAGAAACCGTATTCGAAGGATAAATTTAAGCTGTATTTTCAAAACCCTCAGTTTATGGGTGGCTGGGATAAAGATAAAGAAACCGACTACCGCGCGACGATCTTGCGCTATGGCTCTAAGTACTATTTAGCCATCATGGATAAAAAGTATGCAAAATGCCTCCAGAAAATCGATAAGGATGATGTCAACGGGAACTACGAAAAAATCAACTACAAACTCTTGCCTGGCCCGAATAAAATGCTGCCGAAAGTCTTTTTTTCCAAAAAATGGATGGCCTATTATAACCCTTCCGAAGATATTCAGAAGATTTACAAGAACGGTACCTTTAAAAAAGGTGACATGTTTAATCTGAATGACTGTCACAAGCTTATTGATTTCTTCAAAGATAGTATTTCTCGCTACCCTAAATGGTCAAACGCGTACGATTTTAATTTCTCAGAAACAGAGAAATACAAAGATATTGCGGGGTTTTATCGCGAAGTCGAAGAACAAGGGTATAAGGTGTCGTTCGAATCGGCTTCCAAAAAGGAAGTGGATAAACTTGTTGAAGAAGGCAAACTCTATATGTTCCAGATCTACAACAAAGATTTCTCCGACAAGAGCCATGGCACGCCGAACTTACATACCATGTACTTTAAATTACTGTTCGATGAAAATAATCATGGTCAGATTCGCTTGAGCGGTGGCGCCGAACTGTTTATGCGCCGCGCATCATTGAAAAAGGAAGAACTTGTTGTGCACCCAGCAAACAGTCCGATTGCAAACAAAAATCCGGATAACCCGAAGAAGACTACCACCTTAAGCTATGATGTATATAAAGATAAACGTTTTAGCGAAGATCAGTATGAACTCCACATTCCGATCGCGATTAACAAATGTCCTAAGAACATCTTTAAAATCAATACCGAGGTTCGTGTACTGCTCAAACACGACGACAACCCCTATGTTATTGGCATTGACCGCGGCGAACGCAACCTGCTGTATATTGTCGTTGTGGATGGCAAAGGTAACATTGTTGAACAGTATAGCCTTAATGAAATCATCAATAATTTCAACGGCATTCGTATTAAAACGGACTATCATAGCCTGCTGGACAAAAAAGAGAAAGAACGCTTCGAAGCCCGTCAGAATTGGACCTCTATCGAAAATATCAAGGAACTGAAAGCTGGGTACATTTCTCAAGTGGTCCACAAGATTTGCGAACTGGTGGAAAAGTATGACGCGGTGATCGCACTGGAGGATCTGAATTCGGGCTTTAAAAATAGCCGTGTGAAGGTGGAGAAACAGGTTTACCAAAAGTTCGAAAAAATGCTGATTGACAAACTTAATTACATGGTGGACAAAAAATCTAATCCATGTGCGACCGGCGGGGCTCTCAAAGGCTATCAGATTACGAATAAATTCGAAAGCTTCAAATCCATGTCTACCCAGAACGGATTTATCTTTTACATCCCCGCCTGGCTGACTTCCAAAATTGATCCGAGTACCGGTTTTGTTAATCTGCTGAAAACCAAATATACGTCGATCGCCGATTCAAAAAAATTCATTAGCAGCTTCGACCGTATTATGTACGTCCCGGAAGAAGACCTGTTCGAATTCGCCCTTGACTATAAAAATTTCTCGCGTACGGATGCCGATTATATCAAAAAATGGAAACTGTACAGCTACGGCAACCGCATCCGCATTTTTCGCAACCCAAAGAAGAACAACGTATTTGACTGGGAGGAAGTTTGCTTGACGAGCGCTTACAAAGAGCTCTTTAACAAATATGGGATCAATTATCAGCAAGGGGACATCCGTGCGTTGCTGTGTGAGCAGTCAGATAAAGCGTTCTACAGCTCTTTCATGGCTTTAATGTCGCTGATGTTGCAGATGCGCAATTCGATCACTGGCCGCACCGATGTGGATTTCCTGATTAGCCCGGTCAAAAATAGCGATGGCATTTTTTACGATTCACGTAACTATGAAGCCCAGGAAAACGCTATTCTGCCGAAGAACGCTGATGCGAACGGCGCGTACAATATTGCCCGTAAGGTACTGTGGGCAATTGGTCAGTTCAAGAAAGCGGAAGATGAAAAGTTGGATAAGGTCAAAATTGCGATCTCGAATAAAGAATGGCTGGAATATGCGCAGACCAGCGTGAAACAT
Protein Sequence
MSKLEKFTNCYSLSKTLRFKAIPVGKTQENIDNKRLLVEDEKRAEDYKGVKKLLDRYYLSFINDVLHSIKLKNLNNYISLFRKKTRTEKENKELENLEINLRKEIAKAFKGNEGYKSLFKKDIIETILPEFLDDKDEIALVNSFNGFTTAFTGFFDNRENMFSEEAKSTSIAFRCINENLTRYISNMDIFEKVDAIFDKHEVQEIKEKILNSDYDVEDFFEGEFFNFVLTQEGIDVYNAIIGGFVTESGEKIKGLNEYINLYNQKTKQKLPKFKPLYKQVLSDRESLSFYGEGYTSDEEVLEVFRNTLNKNSEIFSSIKKLEKLFKNFDEYSSAGIFVKNGPAISTISKDIFGEWNVIRDKWNAEYDDIHLKKKAVVTEKYEDDRRKSFKKIGSFSLEQLQEYADADLSVVEKLKEIIIQKVDEIYKVYGSSEKLFDADFVLEKSLKKNDAVVAIMKDLLDSVKSFENYIKAFFGEGKETNRDESFYGDFVLAYDILLKVDHIYDAIRNYVTQKPYSKDKFKLYFQNPQFMGGWDKDKETDYRATILRYGSKYYLAIMDKKYAKCLQKIDKDDVNGNYEKINYKLLPGPNKMLPKVFFSKKWMAYYNPSEDIQKIYKNGTFKKGDMFNLNDCHKLIDFFKDSISRYPKWSNAYDFNFSETEKYKDIAGFYREVEEQGYKVSFESASKKEVDKLVEEGKLYMFQIYNKDFSDKSHGTPNLHTMYFKLLFDENNHGQIRLSGGAELFMRRASLKKEELVVHPANSPIANKNPDNPKKTTTLSYDVYKDKRFSEDQYELHIPIAINKCPKNIFKINTEVRVLLKHDDNPYVIGIDRGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTDYHSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALEDLNSGFKNSRVKVEKQVYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMSTQNGFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFALDYKNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINYQQGDIRALLCEQSDKAFYSSFMALMSLMLQMRNSITGRTDVDFLISPVKNSDGIFYDSRNYEAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQTSVKH
HA HA (human influenza hemagglutinin) epit… 5656..5683 Fwd
DNA Sequence
TATCCCTATGACGTGCCCGATTATGCC
Protein Sequence
YPYDVPDYA
HA HA (human influenza hemagglutinin) epit… 5662..5689 Fwd
DNA Sequence
TATGACGTGCCCGATTATGCCAGCCTG
Protein Sequence
YDVPDYASL
SV40 NLS nuclear localization signal of SV40 (si… 5701..5722 Fwd
DNA Sequence
CCCAAGAAAAAACGCAAGGTG
Protein Sequence
PKKKRKV
SV40 NLS nuclear localization signal of SV40 (si… 5728..5749 Fwd
DNA Sequence
CCTAAGAAAAAGCGGAAAGTG
Protein Sequence
PKKKRKV
mCherry monomeric derivative of DsRed fluoresce… 5788..6493 Fwd
DNA Sequence
GTGAGCAAGGGCGAGGAGGATAACATGGCCATCATCAAGGAGTTCATGCGCTTCAAGGTGCACATGGAGGGCTCCGTGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCCTACGAGGGCACCCAGACCGCCAAGCTGAAGGTGACCAAGGGTGGCCCCCTGCCCTTCGCCTGGGACATCCTGTCCCCTCAGTTCATGTACGGCTCCAAGGCCTACGTGAAGCACCCCGCCGACATCCCCGACTACTTGAAGCTGTCCTTCCCCGAGGGCTTCAAGTGGGAGCGCGTGATGAACTTCGAGGACGGCGGCGTGGTGACCGTGACCCAGGACTCCTCCCTCCAGGACGGCGAGTTCATCTACAAGGTGAAGCTGCGCGGCACCAACTTCCCCTCCGACGGCCCCGTAATGCAGAAGAAGACCATGGGCTGGGAGGCCTCCTCCGAGCGGATGTACCCCGAGGACGGCGCCCTGAAGGGCGAGATCAAGCAGAGGCTGAAGCTGAAGGACGGCGGCCACTACGACGCTGAGGTCAAGACCACCTACAAGGCCAAGAAGCCCGTGCAGCTGCCCGGCGCCTACAACGTCAACATCAAGTTGGACATCACCTCCCACAACGAGGACTACACCATCGTGGAACAGTACGAACGCGCCGAGGGCCGCCACTCCACCGGCGGCATGGACGAGCTGTACAAG
Protein Sequence
VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEGRHSTGGMDELYK
SV40 NLS nuclear localization signal of SV40 (si… 6499..6520 Fwd
DNA Sequence
CCTAAGAAAAAGCGAAAAGTT
Protein Sequence
PKKKRKV
AmpR bla beta-lactamase 7324..8185 Fwd
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGCAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
rop rop Rop protein, which maintains plasmids a… 9370..9562 Rev
DNA Sequence
GTGACCAAACAGGAAAAAACCGCCCTTAACATGGCCCGCTTTATCAGAAGCCAGACATTAACGCTTCTGGAGAAACTCAACGAGCTGGACGCGGATGAACAGGCAGACATCTGTGAATCGCTTCACGACCACGCTGATGAGCTTTACCGCAGCTGCCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGA
Protein Sequence
MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHADELYRSCLARFGDDGENL
promoter 3
Label Gene Product Position Strand
T7 promoter 608..627 Fwd
DNA Sequence
TAATACGACTCACTATAGG
tet promoter 7078..7107 Rev
DNA Sequence
TTGACAGCTTATCATCGATAAGCTTTAAT
AmpR promoter bla 7219..7324 Fwd
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAACATTGAAAAAGGAAGAGT
rep_origin 1
Label Gene Product Position Strand
ori 8355..8944 Fwd
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 1
Label Gene Product Position Strand
T7 terminator 6665..6713 Fwd
DNA Sequence
CTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG