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DsRed-Rab5 WT vector (V001617#)

Basic Vector Information

Basic Vector Information
Vector Name DsRed-Rab5 WT Antibiotic Resistance Kanamycin
Length 5369 bp Type Mammalian Expression
Copy Number High Copy Cloning Method Restriction Enzyme

DsRed-Rab5 WT vector Vector Map

DsRed-Rab5 WT5369 bp6001200180024003000360042004800CMV enhancerCMV promoterKozak sequenceKozak sequenceHAMCSSV40 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 ℃.

DsRed-Rab5 WT vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                5369 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    DsRed-Rab5 WT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5369)
  AUTHORS   Sharma DK, Choudhury A, Singh RD, Wheatley CL, Marks DL, Pagano RE
  TITLE     Glycosphingolipids internalized via caveolar-related endocytosis 
            rapidly merge with the clathrin pathway in early endosomes and form 
            microdomains for recycling.
  JOURNAL   J Biol Chem. 2003 Feb 28. 278(9):7564-72.
  PUBMED    12482757
REFERENCE   2  (bases 1 to 5369)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem. 2003 Feb 28. 278(9):7564-72."
FEATURES             Location/Qualifiers
     source          1..5369
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        16..319
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        320..523
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     474..494
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      562..571
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             568..1242
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(748..768)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1160..1183
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     regulatory      1249..1258
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1261..1287
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    1936..1988
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2112..2233
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2149..2168)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2203..2222
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2240..2695)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2377..2398)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2589..2608
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2722..2826
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2828..3185
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2851..2871)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3036..3171
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3098..3117
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3220..4014
                     /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(3274..3293)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3884..3903
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4202..4221)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4246..4293
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4622..5210
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5111..5130
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"

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