Basic Vector Information
- Vector Name:
- pENTRL2130K-EndoH-TEVH8STREP
- Antibiotic Resistance:
- Kanamycin
- Length:
- 3230 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Mitsudome T, Xu J, Nagata Y, Masuda A, Iiyama K, Morokuma D, Li Z, Mon H, Lee JM, Kusakabe T.
pENTRL2130K-EndoH-TEVH8STREP vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pENTRL2130K-EndoH-TEVH8STREP vector Sequence
LOCUS 40924_17547 3230 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pENTRL2130K-EndoH-TEVH8STREP DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3230)
AUTHORS Mitsudome T, Xu J, Nagata Y, Masuda A, Iiyama K, Morokuma D, Li Z,
Mon H, Lee JM, Kusakabe T.
TITLE Expression, purification, and characterization of
endo-beta-N-acetylglucosaminidase H using baculovirus-mediated
silkworm protein expression system
JOURNAL Appl. Biochem. Biotechnol. 172 (8), 3978-3988 (2014)
PUBMED 24599668
REFERENCE 2 (bases 1 to 3230)
AUTHORS Kusakabe T, Mitsudome T.
TITLE Direct Submission
JOURNAL Submitted (18-JUL-2013) Contact:Takahiro Kusakabe Kyushu University,
Graduate School of Bioresource and Bioenvironmental Sciences,
Laboratory of Silkworm Science; Hakozaki 6-10-1, Fukuoka, Fukuoka
812-8581, Japan
REFERENCE 3 (bases 1 to 3230)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3230)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
Biochem. Biotechnol."; date: "2014"; volume: "172"; issue: "8";
pages: "3978-3988"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(18-JUL-2013) Contact:Takahiro Kusakabe Kyushu University, Graduate
School of Bioresource and Bioenvironmental Sciences, Laboratory of
Silkworm Science; Hakozaki 6-10-1, Fukuoka, Fukuoka 812-8581, Japan"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3230
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 103..189
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 281..308
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
protein_bind 358..456
/label=attL1
/note="recombination site for the Gateway(R) LR reaction"
misc_feature 457..477
/label=L21 enhancer sequence
/note="L21 enhancer sequence"
misc_feature 478..537
/label=Bombyx mori 30K protein signal peptide
/note="Bombyx mori 30K protein signal peptide"
misc_feature 538..1362
/gene="Endo-H-TEVH8STREP"
/label=endo-beta-N-acetylglucosaminidase H-TEVH8STREP
/note="endo-beta-N-acetylglucosaminidase H-TEVH8STREP"
gene 538..1362
/gene="Endo-H-TEVH8STREP"
/label=Endo-H-TEVH8STREP
CDS 610..1362
/codon_start=1
/gene="Endo-H"
/product="endo-beta-N-acetylglucosaminidase H"
/label=Endo-H
/protein_id="BAN66718.1"
/translation="VGKYTLADGGGNAFDVAVIFAANINYDTGTKTAYLHFNENVQRVL
DNAVTQIRPLQQQGIKVLLSVLGNHQGAGFANFPSQQAASAFAKQLSDAVAKYGLDGVD
FDDEYAEYGNNGTAQPNDSSFVHLVTALRANMPDKIISLYNIGPAASRLSYGGVDVSDK
FDYAWNPYYGTWQVPGIALPKAQLSPAAVEIGRTSRSTVADLARRTVDEGYGVYLTYNL
DGGDRTADVSAFTRELYGSEAVRTPSSAG"
gene 610..1362
/gene="Endo-H"
/label=Endo-H
CDS 1363..1383
/codon_start=1
/label=TEV site
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
/translation="ENLYFQG"
CDS 1393..1416
/codon_start=1
/label=8xHis
/note="8xHis affinity tag"
/translation="HHHHHHHH"
CDS 1432..1455
/codon_start=1
/label=Strep-Tag II
/note="peptide that binds Strep-Tactin(R), an engineered
form
of streptavidin"
/translation="WSHPQFEK"
protein_bind complement(1459..1558)
/label=attL2
/note="recombination site for the Gateway(R) LR reaction"
CDS 1681..2487
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 2580..3168
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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