Kif5A-GFP-CIBN vector (V007290)

Basic Vector Information

      • Vector Name:
      • Kif5A-GFP-CIBN
      • Antibiotic Resistance:
      • Kanamycin
      • Length:
      • 6796 bp
      • Type:
      • Mammalian Expression
      • Promoter:
      • CMV
      • Cloning Method:
      • Restriction Enzyme
      • 5' Primer:
      • CMV forward

Kif5A-GFP-CIBN vector Vector Map

Kif5A-GFP-CIBN6796 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600CMV enhancerCMV promoterKozak sequenceEGFPSV40 poly(A) signalf1 oriAmpR promoterSV40 promoterNeoR/KanRTK-pA-RHSV TK poly(A) signalori

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

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                6796 bp ds-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    Kif5A-GFP-CIBN
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)
  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"
FEATURES             Location/Qualifiers
     source          1..6796
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      606..615
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2334..3050
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2334..3050
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2379..2400)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2640..2659)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2987..3008
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3584..3705
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3621..3640)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3675..3694
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3712..4167)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3849..3870)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4061..4080
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4194..4298
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4300..4657
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4323..4343)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4508..4643
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4570..4589
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4692..5486
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4746..4765)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5356..5375
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     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"
     primer_bind     6583..6602
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"

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