Basic Vector Information
EGFP-Rab5A Q79L 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-Rab5A Q79L vector Sequence
LOCUS 40924_725 6748 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 6748) 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 6748) TITLE Direct Submission REFERENCE 3 (bases 1 to 6748) 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..6748 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind complement(46..65) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 237..616 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 617..820 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" promoter 865..883 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 897..1613 /codon_start=1 /label=EGFP /note="enhanced GFP" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" promoter complement(2303..2321) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" polyA_signal 2347..2571 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2617..3045 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 3059..3388 /label=SV40 promoter /note="SV40 enhancer and early promoter" CDS 3455..4246 /codon_start=1 /label=NeoR/KanR /note="aminoglycoside phosphotransferase" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" polyA_signal 4423..4556 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4593..4609) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(4617..4633) /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)." promoter complement(4641..4671) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(4686..4707) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4824..4841) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4995..5583) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(5757..6614) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(6615..6719) /label=AmpR promoter
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