Basic Vector Information
- Vector Name:
- pME-mCherry-T2A-EGFP
- Antibiotic Resistance:
- Kanamycin
- Length:
- 4107 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Hall TE, Currie PD.
pME-mCherry-T2A-EGFP 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 ℃.
pME-mCherry-T2A-EGFP vector Sequence
LOCUS 40924_30510 4107 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pME-mCherry-T2A-EGFP, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4107)
AUTHORS Hall TE, Currie PD.
TITLE Direct Submission
JOURNAL Submitted (27-SEP-2011) Australian Regenerative Medicine Institute,
Monash University, Building 75, Melbourne, VIC 3800, Australia
REFERENCE 2 (bases 1 to 4107)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 4107)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
(27-SEP-2011) Australian Regenerative Medicine Institute, Monash
University, Building 75, Melbourne, VIC 3800, Australia"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4107
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator complement(268..295)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(387..473)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 537..553
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 569..668
/label=attL1
/note="recombination site for the Gateway(R) LR reaction"
CDS 674..1381
/codon_start=1
/label=mCherry
/note="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
misc_feature 1382..1435
/label=T2A viral peptide
/note="T2A viral peptide"
CDS 1382..1435
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 1436..2152
/codon_start=1
/label=EGFP
/note="enhanced GFP"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
misc_feature 2168..2230
/label=CAAX membrane localisation signal (from H-ras)
/note="CAAX membrane localisation signal (from H-ras)"
protein_bind complement(2232..2331)
/label=attL2
/note="recombination site for the Gateway(R) LR reaction"
promoter complement(2349..2367)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2372..2388)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
CDS 2501..3307
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 3457..4045
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
This page is informational only.