Basic Vector Information
- Vector Name:
- Kif5A-GFP-CIBN
- Antibiotic Resistance:
- Kanamycin
- Length:
- 6796 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Promoter:
- CMV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- CMV forward
Kif5A-GFP-CIBN 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 ℃.
Kif5A-GFP-CIBN vector Sequence
LOCUS 40924_1499 6796 bp DNA circular SYN 13-MAY-2021
DEFINITION Expresses fusion of kinesin heavy chain 5A (1-572) with GFP and CIB1
(1-170).
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6796)
AUTHORS Duan L, Che D, Zhang K, Ong Q, Guo S, Cui B
TITLE Optogenetic control of molecular motors and organelle distributions
in cells.
JOURNAL Chem Biol. 2015 May 21;22(5):671-82. doi:
10.1016/j.chembiol.2015.04.014. Epub 2015 May 9.
PUBMED 25963241
REFERENCE 2 (bases 1 to 6796)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6796)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.chembiol.2015.04.014"; journalName: "Chem Biol"; date:
"2015-05-21- 21"; volume: "22"; issue: "5"; pages: "671-82"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6796
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
regulatory 606..615
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 2334..3050
/codon_start=1
/label=EGFP
/note="enhanced GFP"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
polyA_signal 3584..3705
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin complement(3712..4167)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 4194..4298
/label=AmpR promoter
promoter 4300..4657
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 4692..5483
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(5674..5693)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5718..5765
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 6094..6682
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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