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