Basic Vector Information
- Vector Name:
- EGFP-Rab7A Q67L
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6780 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Selection Marker:
- Neomycin (select with G418)
- Copy Number:
- High Copy
- Promoter:
- CMV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- CMV-F
- 3' Primer:
- BGH-rev
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 40924_735 6780 bp DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . 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) TITLE Direct Submission REFERENCE 3 (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." COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..6780 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind 107..128 /label=CAP binding site /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 /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 205..221 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" polyA_signal complement(258..391) /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" CDS complement(568..1359) /codon_start=1 /label=NeoR/KanR /note="aminoglycoside phosphotransferase" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" promoter complement(1426..1755) /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin complement(1769..2197) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" polyA_signal complement(2243..2467) /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" promoter 2493..2511 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" CDS complement(3170..3196) /codon_start=1 /label=HA /note="HA (human influenza hemagglutinin) epitope tag" /translation="YPYDVPDYA" CDS complement(3197..3223) /codon_start=1 /label=HA /note="HA (human influenza hemagglutinin) epitope tag" /translation="YPYDVPDYA" CDS complement(3233..3949) /codon_start=1 /label=EGFP /note="enhanced GFP" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" 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" enhancer complement(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 /label=AmpR promoter CDS 4980..5837 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 6011..6599 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6753..6770 /label=L4440 /note="L4440 vector, forward primer"
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