Lamp1-RFP vector (V007008)

Basic Vector Information

      • Vector Name:
      • Lamp1-RFP
      • Antibiotic Resistance:
      • Kanamycin
      • Length:
      • 5906 bp
      • Type:
      • Mammalian Expression
      • Selection Marker:
      • Neomycin (select with G418)
      • Copy Number:
      • High Copy
      • Cloning Method:
      • Restriction Enzyme
      • 5' Primer:
      • CMV Forward
      • 3' Primer:
      • DsRed1-N

Lamp1-RFP vector Vector Map

Lamp1-RFP5906 bp60012001800240030003600420048005400pBABE 3'AmpR promoterf1 oriSV40 poly(A) signalmRFP1LAMP1CMV promoterCMV enhanceroriHSV TK poly(A) signalTK-pA-RNeoR/KanRSV40 promoter

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 ℃.

Lamp1-RFP vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                5906 bp ds-DNA     circular SYN 21-DEC-2021
DEFINITION  Mammalian expression of Lamp1 fused to RFP. Used for localization to
            the lysosome..
ACCESSION   .
VERSION     .
KEYWORDS    Lamp1-RFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5906)
  AUTHORS   Sherer NM, Lehmann MJ, Jimenez-Soto LF, Ingmundson A, Horner SM, 
            Cicchetti G, Allen PG, Pypaert M, Cunningham JM, Mothes W
  TITLE     Visualization of retroviral replication in living cells reveals 
            budding into multivesicular bodies.
  JOURNAL   Traffic 2003 Nov;4(11):785-801.
  PUBMED    14617360
REFERENCE   2  (bases 1 to 5906)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Traffic"; 
            date: "2003-11"; volume: "4(11)"; pages: "785-801"
FEATURES             Location/Qualifiers
     source          1..5906
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     148..168
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(193..297)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      324..779
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(411..430)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     621..642
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    786..907
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(797..816)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     851..870
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1042..1716)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(1101..1124)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1299..1318)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1568..1586
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1713..1722
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             complement(1738..2958)
                     /codon_start=1
                     /product="rat lysosome-associated membrane glycoprotein 1"
                     /label=LAMP1
                     /translation="MAAPGARRPLLLLLLAGLAHSAPALFEVKDNNGTACIMASFSASF
                     LTTYEAGHVSKVSNMTLPASAEVLKNSSSCGEKNASEPTLAITFGEGYLLKLTFTKNTT
                     RYSVQHMYFTYNLSDTQFFPNASSKGPDTVDSTTDIKADINKTYRCVSDIRVYMKNVTI
                     VLWDATIQAYLPSSNFSKEETRCPQDQPSPTTGPPSPSPPLVPTNPSVSKYNVTGDNGT
                     CLLASMALQLNITYMKKDNTTVTRAFNINPSDKYSGTCGAQLVTLKVGNKSRVLELQFG
                     MNATSSLFFLQGVQLNMTLPDAIEPTFSTSNYSLKALQASVGNSYKCNSEEHIFVSKAL
                     ALNVFSVQVQAFRVESDRFGSVEECVQDGNNMLIPIAVGGALAGLVLIVLIAYLIGRKR
                     SHAGYQTI"
     regulatory      2951..2960
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      2955..2964
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        complement(3033..3236)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3062..3082)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3237..3540
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3715..4303)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3795..3814)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4632..4679
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4704..4723
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4911..5705)
                     /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(5022..5041)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5632..5651
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5740..5906,1..191))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5754..5889
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5808..5827)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"

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