pDEST-hisMBP-AsCpf1-EC

79007 Synthetic Bacterial Expression

Plasmid Information

Purpose
Expressing his-MBP tagged AsCpf1 (E.coli codon optimized)
Depositor
Jin-Soo Kim
Article
Hur et al Nat Biotechnol. 2016 Jun 6. doi: 10.1038
Insert
AsCpf1
Tags
6xHis-MBP-NLS and NLS-HA
Promoter
tac promoter
Available Since
July 22, 2016
Availability
Available

Sequence Information

Length
10,644 bp
Topology
circular
Molecule Type
other DNA

Features

CDS 8
Label Gene Product Position Strand
6xHis — 6xHis affinity tag 131..149 Fwd
DNA Sequence
CATCACCATCACCATCAC
Protein Sequence
HHHHHH
SV40 NLS — nuclear localization signal of SV40 (si… 1289..1310 Fwd
DNA Sequence
CCAAAAAAGAAGAGAAAGGTA
Protein Sequence
PKKKRKV
AsCpf1 — CRISPR-associated protein from Acidamin… 1316..5237 Fwd
DNA Sequence
ATGACACAGTTTGAAGGCTTCACCAATCTCTACCAGGTCAGCAAGACGCTACGTTTTGAGCTTATCCCGCAGGGAAAAACCCTGAAACACATTCAGGAACAGGGGTTCATAGAGGAAGATAAGGCGCGTAACGACCATTATAAAGAACTGAAGCCTATAATCGACCGTATTTATAAAACGTACGCGGATCAGTGCCTGCAGCTGGTTCAGCTGGATTGGGAGAATCTGTCCGCGGCTATTGATAGCTATCGCAAAGAGAAGACCGAGGAAACCCGTAACGCACTGATTGAAGAGCAGGCGACCTATCGGAATGCGATCCATGATTACTTCATCGGCCGCACCGACAACCTGACCGATGCAATTAACAAACGTCACGCAGAGATTTACAAAGGTCTGTTTAAAGCAGAGTTATTCAATGGCAAGGTTCTGAAACAGCTGGGTACGGTCACCACCACCGAACACGAAAACGCACTGCTGAGGAGCTTTGATAAATTTACCACATATTTCAGCGGTTTCTATGAAAATCGTAAGAATGTATTTAGCGCCGAAGATATTTCCACCGCAATTCCTCATCGTATTGTGCAGGATAATTTTCCGAAGTTTAAAGAAAATTGTCATATTTTTACCCGTCTGATCACCGCGGTACCGAGCCTGCGAGAGCATTTTGAAAACGTTAAGAAAGCCATTGGAATTTTTGTCAGTACCAGCATTGAAGAAGTGTTTTCGTTCCCGTTCTATAACCAACTGCTGACCCAGACCCAGATTGATCTGTACAATCAGCTGCTGGGGGGCATAAGCCGCGAGGCAGGTACCGAAAAGATAAAGGGACTCAATGAGGTGCTGAATCTGGCAATTCAGAAGAATGATGAAACGGCTCATATCATTGCTAGCCTGCCGCATCGTTTCATTCCCCTGTTTAAGCAAATCCTGAGCGATCGCAATACACTGAGCTTTATCCTCGAAGAGTTTAAATCGGACGAAGAAGTTATCCAGAGCTTTTGCAAATACAAAACCCTGCTGCGGAACGAAAATGTGCTGGAGACCGCTGAAGCACTGTTTAATGAACTGAACTCGATCGACCTCACCCATATTTTTATATCCCACAAAAAACTGGAAACCATAAGCAGCGCTCTGTGTGACCATTGGGATACCCTGCGCAACGCCCTGTATGAACGGCGTATCAGCGAGCTGACCGGGAAAATCACCAAATCCGCAAAGGAAAAAGTTCAGCGTAGTCTGAAACACGAGGACATCAACCTGCAAGAAATTATTAGCGCAGCAGGTAAAGAGCTGAGCGAAGCATTCAAACAGAAAACCAGCGAAATCCTGAGCCATGCCCATGCTGCACTGGATCAGCCGCTGCCGACCACCCTGAAAAAACAGGAGGAAAAGGAGATTCTGAAAAGCCAACTGGACAGCCTGCTGGGCCTGTATCACCTGCTGGACTGGTTTGCAGTCGATGAGAGCAACGAGGTTGATCCTGAGTTCTCCGCTCGTCTGACCGGAATCAAGCTGGAGATGGAACCGAGTCTGTCGTTTTACAATAAAGCGCGTAATTACGCGACCAAGAAACCGTATAGCGTGGAAAAATTCAAACTGAACTTTCAGATGCCGACCCTTGCAAGCGGATGGGACGTTAACAAAGAAAAAAACAATGGGGCAATTCTGTTTGTGAAAAATGGCCTCTATTATCTGGGTATCATGCCGAAACAGAAAGGGCGCTACAAAGCCCTGTCATTTGAGCCGACCGAGAAAACCTCAGAGGGTTTCGACAAGATGTACTACGATTATTTCCCGGATGCGGCAAAAATGATACCCAAATGTAGCACCCAACTGAAGGCAGTTACAGCCCACTTTCAGACCCATACCACCCCGATCCTGCTGTCGAACAATTTTATAGAGCCGCTGGAAATTACCAAAGAGATTTATGATCTGAATAATCCGGAAAAGGAGCCCAAGAAATTTCAGACGGCGTATGCAAAAAAGACCGGGGATCAGAAAGGTTATCGTGAAGCGCTGTGCAAATGGATTGACTTTACCCGTGACTTTCTGTCAAAATATACCAAAACGACGAGCATTGATCTGAGCAGCCTACGTCCGAGCAGCCAATATAAGGATCTGGGCGAATATTACGCCGAACTGAATCCGCTGCTCTACCATATTTCCTTCCAACGAATCGCTGAAAAAGAAATAATGGACGCCGTTGAAACCGGCAAACTGTATCTGTTTCAAATCTACAACAAAGATTTCGCCAAAGGCCATCACGGTAAGCCGAACCTGCATACCCTGTATTGGACCGGTCTGTTTAGCCCGGAGAATCTGGCCAAAACCAGCATCAAGCTGAACGGACAGGCAGAACTGTTTTACCGCCCCAAAAGCCGTATGAAAAGGATGGCACACCGCCTGGGCGAAAAAATGCTGAATAAGAAACTCAAAGATCAGAAAACGCCGATACCGGATACCCTTTATCAGGAGCTGTATGATTATGTTAACCACCGGCTGAGCCATGACCTGAGCGACGAAGCGCGTGCACTGCTGCCGAACGTGATTACCAAGGAAGTCTCGCATGAAATTATTAAAGATCGGCGCTTCACCAGTGATAAATTTTTCTTCCATGTACCGATCACCCTGAATTATCAAGCCGCAAATAGCCCTTCCAAATTTAATCAACGCGTGAATGCGTACCTGAAAGAGCATCCGGAGACCCCAATTATTGGCATAGACCGAGGAGAACGCAATCTCATTTATATCACCGTCATTGATAGCACCGGTAAGATCCTGGAACAGCGTAGCCTGAATACCATTCAGCAGTTTGACTACCAGAAAAAGCTGGACAACAGAGAAAAGGAACGTGTAGCCGCCCGGCAGGCTTGGAGTGTGGTGGGTACTATCAAGGATCTGAAGCAGGGGTATCTCTCCCAAGTTATCCATGAAATTGTCGATCTAATGATTCACTATCAAGCAGTAGTGGTACTGGAAAATCTGAATTTCGGTTTCAAAAGCAAACGTACAGGGATCGCTGAAAAAGCCGTTTATCAGCAGTTCGAGAAAATGCTGATAGACAAGCTGAATTGCCTGGTTCTGAAAGATTATCCGGCAGAGAAGGTGGGCGGTGTGCTGAACCCGTACCAGCTGACTGATCAATTTACGAGCTTTGCAAAAATGGGAACGCAGAGCGGTTTCCTGTTCTATGTTCCGGCGCCATATACCAGCAAGATAGACCCGCTGACAGGTTTCGTAGATCCGTTTGTCTGGAAAACCATTAAAAATCATGAAAGTCGCAAACATTTTCTGGAGGGCTTTGATTTTCTGCACTATGACGTGAAAACCGGCGACTTCATTCTGCATTTTAAAATGAACCGTAATCTGTCCTTTCAGCGCGGCCTGCCTGGCTTTATGCCGGCGTGGGACATTGTTTTTGAAAAGAATGAGACACAGTTTGATGCCAAAGGTACCCCCTTTATTGCGGGGAAACGCATTGTGCCCGTTATAGAAAATCACCGCTTCACCGGACGGTATAGGGACTTGTACCCGGCAAATGAATTGATAGCGCTGCTGGAGGAGAAAGGTATTGTCTTTCGGGATGGATCAAACATCCTGCCGAAGCTGCTGGAGAACGATGACAGCCACGCAATAGACACCATGGTAGCGCTGATCCGAAGCGTGCTGCAGATGCGTAACAGTAATGCGGCTACGGGGGAAGACTACATTAATAGCCCGGTCCGTGATCTGAACGGCGTTTGTTTCGATAGCAGATTTCAAAATCCGGAGTGGCCGATGGATGCCGATGCCAATGGAGCTTACCATATCGCTCTCAAAGGTCAGCTCCTACTGAACCATTTGAAAGAATCAAAAGATCTGAAACTGCAGAACGGCATCTCGAATCAGGACTGGCTGGCCTACATTCAAGAACTGAGAAAC
Protein Sequence
MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDHYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNAIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDKFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLNLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLRNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGKITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLKKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKARNYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYKALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPLEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDLSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGHHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKDQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFHVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRSLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVVVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQFTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDVKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENHRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMRNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESKDLKLQNGISNQDWLAYIQELRN
SV40 NLS — nuclear localization signal of SV40 (si… 5237..5258 Fwd
DNA Sequence
CCAAAGAAAAAGAGAAAAGTA
Protein Sequence
PKKKRKV
HA — HA (human influenza hemagglutinin) epit… 5264..5291 Fwd
DNA Sequence
TACCCCTACGACGTGCCCGACTACGCC
Protein Sequence
YPYDVPDYA
AmpR bla beta-lactamase 6012..6873 Fwd
DNA Sequence
ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTCCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
rop rop Rop protein, which maintains plasmids a… 8554..8746 Rev
DNA Sequence
GTGACCAAACAGGAAAAAACCGCCCTTAACATGGCCCGCTTTATCAGAAGCCAGACATTAACGCTTCTGGAGAAACTCAACGAGCTGGACGCGGATGAACAGGCAGACATCTGTGAATCGCTGCACGACCACGCTGATGAGCTTTACCGCAGCTGCCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGA
Protein Sequence
MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHADELYRSCLARFGDDGENL
lacI lacI lac repressor 9319..10402 Fwd
DNA Sequence
GTGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGA
Protein Sequence
MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ
promoter 3
Label Gene Product Position Strand
tac promoter — — 0..29 Fwd
DNA Sequence
TTGACAATTAATCATCGGCTCGTATAATG
AmpR promoter bla — 5920..6012 Fwd
DNA Sequence
TTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
lacIq promoter lacI (mutant) — 9241..9319 Fwd
DNA Sequence
GACACCATCGAATGGTGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAAT
protein_bind 3
Label Gene Product Position Strand
lac operator — — 36..53 Fwd
DNA Sequence
TTGTGAGCGGATAACAA
attB1 mutant version of attB — 1250..1275 Fwd
DNA Sequence
ACAAGTTTGTACAAAAAAGCAGGCT
attB2 mutant version of attB — 5295..5320 Rev
DNA Sequence
ACCACTTTGTACAAGAAAGCTGGGT
rep_origin 2
Label Gene Product Position Strand
M13 ori — — 6914..7428 Rev
DNA Sequence
ACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTAAATATTTGCTTATACAATCTTCCTGTTTTTGGGGCTTTTCTGATTATCAACCGGGGT
ori — — 7538..8127 Fwd
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 2
Label Gene Product Position Strand
rrnB T1 terminator Escherichia coli rrnB — 5696..5783 Fwd
DNA Sequence
CAAATAAAACGAAAGGCTCAGTCGAAAGACTGGGCCTTTCGTTTTATCTGTTGTTTGTCGGTGAACGCTCTCCTGAGTAGGACAAAT
rrnB T2 terminator — — 5874..5902 Fwd
DNA Sequence
AGAAGGCCATCCTGACGGATGGCCTTTT