Basic Vector Information
EGFP-Rab7A Q67L 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 ℃.
EGFP-Rab7A Q67L vector Sequence
LOCUS Exported 6780 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS EGFP-Rab7A Q67L
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6780)
AUTHORS Sun Q, Westphal W, Wong KN, Tan I, Zhong Q
TITLE Rubicon controls endosome maturation as a Rab7 effector.
JOURNAL Proc Natl Acad Sci U S A. 2010 Nov 9. 107(45):19338-43.
PUBMED 20974968
REFERENCE 2 (bases 1 to 6780)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl
Acad Sci U S A. 2010 Nov 9. 107(45):19338-43."
FEATURES Location/Qualifiers
source 1..6780
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 186..208
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 205..221
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
polyA_signal 270..391
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(281..300)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 335..354
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(565..1359)
/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
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(676..695)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 1286..1305
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
promoter complement(1426..1755)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 1440..1575
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(1494..1513)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 1740..1760
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin complement(1769..2197)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1879..1900)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2091..2110
/label=F1ori-R
/note="F1 origin, reverse primer"
polyA_signal 2243..2467
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 2456..2473
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
promoter 2493..2511
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 2493..2510
/label=SP6
/note="SP6 promoter, forward primer"
CDS complement(3170..3196)
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS complement(3197..3223)
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS complement(3233..3949)
/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 complement(3233..3949)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3275..3296)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 3624..3643
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3883..3904
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3962..3981)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3963..3981)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(4026..4229)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(4055..4075)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 4230..4609
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind 4781..4800
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 4875..4979
/gene="bla"
/label=AmpR promoter
CDS 4980..5840
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(5198..5217)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6011..6599
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6500..6519
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6753..6770
/label=L4440
/note="L4440 vector, forward primer"
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