Basic Vector Information
EGFP-Rab7A 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 vector Sequence
LOCUS Exported 6780 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS EGFP-Rab7A 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" primer_bind complement(42..61) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 233..612 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 613..816 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 767..787 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 861..880 /label=T7 /note="T7 promoter, forward primer" promoter 861..879 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 893..1609 /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 893..1609 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(938..959) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1199..1218) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1546..1567 /label=EGFP-C /note="EGFP, forward primer" CDS 1619..1645 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 1646..1672 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" promoter complement(2331..2349) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2332..2349) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2369..2386) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2375..2599 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2645..3073 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2732..2751) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2942..2963 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3082..3102) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3087..3416 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3267..3402 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3329..3348 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3483..4277 /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(3537..3556) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4147..4166 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4451..4572 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4488..4507) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4542..4561 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4621..4637) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4621..4637) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4634..4656) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4645..4661 /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)." promoter complement(4669..4699) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4714..4735 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4852..4869) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5023..5611) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5103..5122) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5782..6642) /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 6405..6424 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6643..6747) /gene="bla" /label=AmpR promoter
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