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EGFP-Rab5A Q79L vector (V001228#)

Basic Vector Information

Basic Vector Information
Vector Name EGFP-Rab5A Q79L Antibiotic Resistance Ampicillin
Length 6748 bp Type Mammalian Expression
Selection Marker Neomycin (select with G418) Copy Number High Copy
Cloning Method Restriction Enzyme 5' Primer CMV-F
3' Primer BGH-rev

EGFP-Rab5A Q79L vector Vector Map

EGFP-Rab5A Q79L6748 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600pRS-markerCMV enhancerCMV promoterT7EGFPSP6 promoterBGH-revf1 oripBABE 3'NeoR/KanRSV40 poly(A) signalM13 revlac operatorlac promoterCAP binding siteL4440oriAmpRAmpR 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 ℃.

EGFP-Rab5A Q79L vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                6748 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
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
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..6748
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(46..65)
                     /note="pRS vectors, use to sequence yeast selectable 
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
     primer_bind     771..791
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             897..1613
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /note="mammalian codon-optimized"
     CDS             897..1613
                     /product="enhanced GFP"
     primer_bind     complement(942..963)
                     /note="EGFP, reverse primer"
     primer_bind     complement(1203..1222)
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1550..1571
                     /note="EGFP, forward primer"
     promoter        complement(2303..2321)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2304..2321)
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2341..2358)
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     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"
     primer_bind     complement(2704..2723)
                     /note="F1 origin, reverse primer"
     primer_bind     2914..2935
                     /note="F1 origin, forward primer"
     primer_bind     complement(3054..3074)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3059..3388
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3239..3374
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3301..3320
                     /note="SV40 promoter/origin, forward primer"
     CDS             3455..4249
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="confers resistance to neomycin, kanamycin, and G418 
     primer_bind     complement(3509..3528)
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4119..4138
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4423..4544
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4460..4479)
                     /note="SV40 polyA, reverse primer"
     primer_bind     4514..4533
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4593..4609)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
     primer_bind     complement(4593..4609)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4606..4628)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4617..4633
                     /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(4641..4671)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4686..4707
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4824..4841)
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4995..5583)
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
     primer_bind     complement(5075..5094)
                     /note="pBR322 origin, forward primer"
     CDS             complement(5754..6614)
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     primer_bind     6377..6396
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6615..6719)
                     /label=AmpR promoter

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