AsCpf1-2NLS

102565 Synthetic Bacterial Expression

Plasmid Information

Purpose
bacterial expression of AsCpf1-2NLS
Depositor
Jennifer Doudna
Article
Moreno-Mateos et al Nat Commun. 2017 Dec 8;8(1):2024. doi: 1
Insert
AsCpf1
Tags
6xHis-MBP-TEV and HA-2xSV40NLS
Promoter
T7
Available Since
Dec. 21, 2017
Availability
Available

Sequence Information

Length
10,751 bp
Topology
circular
Molecule Type
other DNA

Features

CDS 12
Label Gene Product Position Strand
rop rop Rop protein, which maintains plasmids a… 384..576 Rev
DNA Sequence
GTGACCAAACAGGAAAAAACCGCCCTTAACATGGCCCGCTTTATCAGAAGCCAGACATTAACGCTTCTGGAGAAACTCAACGAGCTGGACGCGGATGAACAGGCAGACATCTGTGAATCGCTTCACGACCACGCTGATGAGCTTTACCGCAGCTGCCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGA
Protein Sequence
MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHADELYRSCLARFGDDGENL
6xHis 6xHis affinity tag 2285..2303 Fwd
DNA Sequence
CACCATCACCATCACCAT
Protein Sequence
HHHHHH
MBP malE (mutated) maltose binding protein from E. coli 2312..3413 Fwd
DNA Sequence
ATGAAAATCGAAGAAGGTAAACTGGTAATCTGGATTAACGGCGATAAAGGCTATAACGGTCTCGCTGAAGTCGGTAAGAAATTCGAGAAAGATACCGGAATTAAAGTCACCGTTGAGCATCCGGATAAACTGGAAGAGAAATTCCCACAGGTTGCGGCAACTGGCGATGGCCCTGACATTATCTTCTGGGCACACGACCGCTTTGGTGGCTACGCTCAATCTGGCCTGTTGGCTGAAATCACCCCGGACAAAGCGTTCCAGGACAAGCTGTATCCGTTTACCTGGGATGCCGTACGTTACAACGGCAAGCTGATTGCTTACCCGATCGCTGTTGAAGCGTTATCGCTGATTTATAACAAAGATCTGCTGCCGAACCCGCCAAAAACCTGGGAAGAGATCCCGGCGCTGGATAAAGAACTGAAAGCGAAAGGTAAGAGCGCGCTGATGTTCAACCTGCAAGAACCGTACTTCACCTGGCCGCTGATTGCTGCTGACGGGGGTTATGCGTTCAAGTATGAAAACGGCAAGTACGACATTAAAGACGTGGGCGTGGATAACGCTGGCGCGAAAGCGGGTCTGACCTTCCTGGTTGACCTGATTAAAAACAAACACATGAATGCAGACACCGATTACTCCATCGCAGAAGCTGCCTTTAATAAAGGCGAAACAGCGATGACCATCAACGGCCCGTGGGCATGGTCCAACATCGACACCAGCAAAGTGAATTATGGTGTAACGGTACTGCCGACCTTCAAGGGTCAACCATCCAAACCGTTCGTTGGCGTGCTGAGCGCAGGTATTAACGCCGCCAGTCCGAACAAAGAGCTGGCAAAAGAGTTCCTCGAAAACTATCTGCTGACTGATGAAGGTCTGGAAGCGGTTAATAAAGACAAACCGCTGGGTGCCGTAGCGCTGAAGTCTTACGAGGAAGAGTTGGCGAAAGATCCACGTATTGCCGCCACTATGGAAAACGCCCAGAAAGGTGAAATCATGCCGAACATCCCGCAGATGTCCGCTTTCTGGTATGCCGTGCGTACTGCGGTGATCAACGCCGCCAGCGGTCGTCAGACTGTCGATGAAGCCCTGAAAGACGCGCAGACT
Protein Sequence
MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQT
TEV site tobacco etch virus (TEV) protease recog… 3467..3488 Fwd
DNA Sequence
GAAAACCTGTACTTCCAATCC
Protein Sequence
ENLYFQS
AsCpf1 CRISPR-associated protein from Acidamin… 3497..7418 Fwd
DNA Sequence
ATGACCCAGTTCGAGGGGTTTACCAATCTGTATCAAGTGAGCAAGACGCTGCGCTTTGAACTGATCCCACAGGGAAAAACCTTAAAACATATTCAAGAGCAGGGCTTTATCGAAGAAGATAAGGCCCGTAATGACCATTACAAAGAGTTAAAGCCGATTATTGATCGTATCTACAAGACCTATGCGGACCAGTGCTTACAATTGGTACAGCTTGATTGGGAGAACCTCTCTGCCGCCATCGATTCCTATCGTAAAGAAAAAACTGAAGAAACGCGCAACGCCCTGATTGAAGAGCAGGCCACCTATCGTAACGCGATTCATGACTATTTTATTGGCCGTACGGACAATCTGACGGACGCGATCAACAAGCGCCATGCGGAGATTTACAAAGGACTGTTTAAGGCTGAACTGTTCAATGGTAAGGTCCTTAAACAGCTTGGGACCGTCACAACGACGGAACATGAAAACGCGTTATTACGTAGCTTCGACAAGTTTACCACGTATTTCTCCGGCTTTTACGAAAATCGCAAAAACGTTTTCAGTGCCGAGGATATTTCCACTGCTATCCCTCATCGCATTGTGCAAGACAACTTCCCAAAATTCAAAGAAAATTGTCATATCTTCACCCGCTTAATCACCGCTGTACCGTCCCTGCGTGAGCATTTCGAAAACGTGAAAAAGGCCATTGGTATCTTCGTGTCTACTTCGATTGAGGAGGTATTTTCCTTTCCATTCTATAATCAGCTGCTGACCCAGACCCAAATTGATCTGTACAACCAGCTGCTTGGCGGTATTTCTCGTGAAGCAGGAACCGAAAAAATCAAAGGGTTGAACGAGGTGCTTAATCTGGCAATCCAGAAAAATGATGAAACCGCCCACATCATTGCTTCGTTACCTCATCGTTTTATCCCGTTGTTCAAGCAAATTTTAAGTGATCGCAATACGCTGTCGTTTATTCTGGAAGAATTCAAAAGTGATGAAGAGGTAATTCAGTCGTTTTGCAAATATAAAACCCTGTTACGTAACGAAAATGTCCTGGAAACAGCCGAGGCTTTGTTTAACGAACTGAATAGCATTGACCTGACGCATATCTTTATTAGCCACAAAAAATTAGAGACCATCTCATCAGCTCTGTGCGATCATTGGGATACACTGCGCAATGCGCTGTATGAACGTCGTATTTCGGAATTGACTGGCAAAATCACTAAAAGCGCGAAAGAGAAAGTACAGCGCTCGCTTAAACATGAAGATATCAACCTGCAGGAGATCATCAGCGCCGCGGGTAAAGAACTGTCGGAGGCATTTAAACAGAAGACGAGCGAGATTCTGTCCCACGCACATGCCGCCTTAGACCAGCCGCTCCCGACCACTCTGAAGAAACAGGAAGAGAAAGAAATCCTTAAAAGTCAACTGGACAGTTTACTGGGTCTCTATCATCTGCTGGATTGGTTTGCGGTAGACGAAAGCAATGAAGTGGATCCGGAGTTTAGTGCCCGTCTGACAGGAATCAAGCTGGAAATGGAGCCTTCGCTTAGCTTCTACAACAAAGCCCGCAATTATGCCACGAAAAAACCCTATAGTGTCGAAAAATTTAAACTCAACTTTCAAATGCCGACCCTTGCGTCGGGCTGGGATGTCAACAAAGAAAAAAACAACGGAGCTATTCTGTTCGTTAAAAATGGTCTGTACTACCTGGGCATCATGCCGAAACAGAAAGGTCGCTACAAAGCCCTTTCGTTCGAGCCCACGGAAAAAACAAGCGAAGGCTTCGACAAAATGTACTACGATTACTTTCCGGATGCAGCAAAAATGATCCCGAAATGTTCCACACAGCTGAAAGCCGTTACAGCACATTTTCAGACGCACACCACCCCCATCTTACTGTCCAACAATTTTATTGAACCGCTGGAGATTACTAAAGAAATTTATGATTTGAACAATCCGGAAAAAGAGCCAAAAAAGTTTCAAACCGCCTACGCTAAAAAAACCGGGGATCAGAAAGGGTACCGCGAAGCGTTGTGCAAGTGGATTGATTTCACCCGCGATTTTCTCAGTAAATATACCAAGACTACCTCGATTGACCTGAGCTCACTGCGCCCGAGCTCTCAATATAAGGATTTGGGTGAGTACTATGCTGAATTAAACCCTTTATTGTACCACATTTCTTTTCAGCGCATCGCCGAAAAGGAAATTATGGACGCAGTCGAAACCGGGAAACTGTACCTGTTCCAGATCTATAATAAGGACTTCGCCAAAGGACATCATGGCAAACCGAACCTGCACACCCTTTACTGGACCGGGCTTTTCTCTCCGGAAAATTTGGCGAAAACCTCGATCAAGCTTAACGGTCAAGCTGAGCTGTTTTACCGTCCAAAATCCCGCATGAAGCGCATGGCGCATCGTTTAGGTGAAAAAATGCTGAATAAGAAACTGAAAGATCAGAAAACCCCTATCCCGGATACCCTCTACCAGGAACTGTATGATTACGTGAACCATCGTCTCTCGCATGACCTGTCAGACGAAGCGCGTGCGTTACTGCCCAATGTAATCACAAAAGAAGTTTCGCATGAAATTATTAAAGATCGTCGTTTTACATCTGATAAATTCTTTTTTCATGTTCCGATCACCCTCAACTATCAGGCCGCAAACAGTCCAAGTAAGTTTAACCAGCGCGTTAATGCTTACCTGAAGGAACATCCGGAGACTCCGATTATTGGAATTGATCGCGGTGAACGTAATTTGATCTATATCACTGTGATCGATAGTACCGGTAAGATTCTGGAGCAGCGCAGCTTGAACACAATTCAACAGTTTGATTATCAGAAAAAATTAGACAACCGCGAAAAAGAGCGCGTGGCTGCCCGTCAGGCGTGGTCTGTTGTCGGTACCATTAAAGATCTGAAGCAGGGCTATCTTTCTCAGGTTATTCACGAAATTGTAGATCTGATGATCCATTATCAGGCGGTTGTTGTGTTGGAGAATCTCAATTTCGGTTTTAAGAGTAAGCGCACAGGCATCGCTGAAAAAGCAGTTTATCAGCAGTTTGAAAAAATGCTGATCGACAAATTGAACTGTTTAGTTCTCAAAGATTACCCAGCGGAAAAGGTGGGCGGAGTGCTGAATCCGTACCAATTAACGGATCAATTCACTTCCTTCGCAAAGATGGGTACCCAAAGCGGCTTTCTGTTCTATGTGCCGGCCCCGTATACCTCGAAAATCGATCCACTGACGGGCTTCGTAGATCCGTTCGTGTGGAAAACCATTAAAAATCATGAAAGTCGTAAACATTTTCTCGAAGGCTTCGACTTCCTGCACTACGACGTGAAAACTGGCGATTTCATTCTGCATTTTAAAATGAACCGCAACCTTTCGTTTCAGCGCGGTCTGCCGGGCTTTATGCCGGCTTGGGACATTGTTTTTGAGAAAAATGAAACCCAGTTTGATGCTAAAGGCACTCCTTTCATCGCCGGTAAACGCATCGTACCTGTGATTGAAAACCATCGTTTTACAGGGCGTTACCGTGATTTATACCCGGCGAACGAATTGATCGCGCTGCTGGAGGAAAAGGGCATCGTTTTCCGTGACGGCTCCAATATTCTGCCGAAATTACTGGAAAACGACGATTCACACGCAATTGATACCATGGTCGCACTGATTCGCTCAGTCTTACAGATGCGTAACTCTAATGCAGCCACAGGAGAAGATTATATTAATTCGCCAGTCCGCGATTTGAACGGTGTTTGCTTCGACAGCCGTTTTCAGAATCCTGAATGGCCGATGGACGCTGATGCCAACGGAGCTTATCATATCGCCCTGAAAGGCCAGCTCCTGCTGAACCACCTGAAGGAAAGCAAAGATCTGAAATTGCAGAACGGCATTAGCAACCAGGACTGGTTAGCATACATCCAGGAACTGCGTAAC
Protein Sequence
MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDHYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNAIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDKFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLNLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLRNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGKITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLKKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKARNYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYKALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPLEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDLSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGHHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKDQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFHVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRSLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVVVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQFTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDVKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENHRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMRNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESKDLKLQNGISNQDWLAYIQELRN
HA HA (human influenza hemagglutinin) epit… 7421..7448 Fwd
DNA Sequence
TATCCCTATGACGTGCCCGATTATGCC
Protein Sequence
YPYDVPDYA
HA HA (human influenza hemagglutinin) epit… 7427..7454 Fwd
DNA Sequence
TATGACGTGCCCGATTATGCCAGCCTG
Protein Sequence
YDVPDYASL
SV40 NLS nuclear localization signal of SV40 (si… 7466..7487 Fwd
DNA Sequence
CCCAAGAAAAAACGCAAGGTG
Protein Sequence
PKKKRKV
SV40 NLS nuclear localization signal of SV40 (si… 7493..7514 Fwd
DNA Sequence
CCTAAGAAAAAGCGGAAAGTG
Protein Sequence
PKKKRKV
mCherry monomeric derivative of DsRed fluoresce… 7553..8258 Fwd
DNA Sequence
GTGAGCAAGGGCGAGGAGGATAACATGGCCATCATCAAGGAGTTCATGCGCTTCAAGGTGCACATGGAGGGCTCCGTGAACGGCCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCCTACGAGGGCACCCAGACCGCCAAGCTGAAGGTGACCAAGGGTGGCCCCCTGCCCTTCGCCTGGGACATCCTGTCCCCTCAGTTCATGTACGGCTCCAAGGCCTACGTGAAGCACCCCGCCGACATCCCCGACTACTTGAAGCTGTCCTTCCCCGAGGGCTTCAAGTGGGAGCGCGTGATGAACTTCGAGGACGGCGGCGTGGTGACCGTGACCCAGGACTCCTCCCTCCAGGACGGCGAGTTCATCTACAAGGTGAAGCTGCGCGGCACCAACTTCCCCTCCGACGGCCCCGTAATGCAGAAGAAGACCATGGGCTGGGAGGCCTCCTCCGAGCGGATGTACCCCGAGGACGGCGCCCTGAAGGGCGAGATCAAGCAGAGGCTGAAGCTGAAGGACGGCGGCCACTACGACGCTGAGGTCAAGACCACCTACAAGGCCAAGAAGCCCGTGCAGCTGCCCGGCGCCTACAACGTCAACATCAAGTTGGACATCACCTCCCACAACGAGGACTACACCATCGTGGAACAGTACGAACGCGCCGAGGGCCGCCACTCCACCGGCGGCATGGACGAGCTGTACAAG
Protein Sequence
VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEGRHSTGGMDELYK
SV40 NLS nuclear localization signal of SV40 (si… 8264..8285 Fwd
DNA Sequence
CCTAAGAAAAAGCGAAAAGTT
Protein Sequence
PKKKRKV
AmpR bla beta-lactamase 9089..9950 Fwd
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGCAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
promoter 3
Label Gene Product Position Strand
T7 promoter 2136..2155 Fwd
DNA Sequence
TAATACGACTCACTATAGG
tet promoter 8843..8872 Rev
DNA Sequence
TTGACAGCTTATCATCGATAAGCTTTAAT
AmpR promoter bla 8984..9089 Fwd
DNA Sequence
CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAACATTGAAAAAGGAAGAGT
rep_origin 1
Label Gene Product Position Strand
ori 10120..10709 Fwd
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 1
Label Gene Product Position Strand
T7 terminator 8430..8478 Fwd
DNA Sequence
CTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG