Basic Vector Information
pyEF1A 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 ℃.
pyEF1A vector Sequence
LOCUS 40924_48008 5324 bp DNA circular SYN 18-DEC-2018
DEFINITION RpL23-fusion expression vector pyEF1A, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5324)
AUTHORS Sorensen HP, Kristensen JE, Sperling-Petersen HU, Mortensen KK.
TITLE Soluble expression of aggregating proteins by covalent coupling to
the ribosome
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5324)
AUTHORS Sorensen HP, Sperling-Petersen HU, Mortensen KK.
TITLE Direct Submission
JOURNAL Submitted (29-MAR-2004) Department of Molecular Biology, Aarhus
University, Gustav Wieds Vej 10C, Aarhus C 8000, Denmark
REFERENCE 3 (bases 1 to 5324)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5324)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(29-MAR-2004) Department of Molecular Biology, Aarhus University,
Gustav Wieds Vej 10C, Aarhus C 8000, Denmark"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5324
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(78..734)
/label=CmR
/note="chloramphenicol acetyltransferase"
promoter complement(735..837)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
rep_origin complement(1363..1907)
/direction=LEFT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin."
CDS complement(2049..2924)
/label=araC
/note="L-arabinose regulatory protein"
promoter 2951..3235
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
RBS 3262..3284
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 3292..3591
/gene="rplW"
/label=rplW
/note="Large ribosomal subunit protein uL23 from
Escherichia coli (strain K12). Accession#: P0ADZ0"
CDS 3592..3603
/label=Factor Xa site
/note="Factor Xa recognition and cleavage site"
CDS 3604..3624
/label=TEV site
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
CDS 3628..5001
/gene="TEF1"
/label=TEF1
/note="Elongation factor 1-alpha from Saccharomyces
cerevisiae (strain ATCC 204508 / S288c). Accession#:
P02994"
CDS 5002..5019
/label=6xHis
/note="6xHis affinity tag"
terminator 5250..5296
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
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