Basic Vector Information
PRExpress 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 ℃.
PRExpress vector Sequence
LOCUS 40924_36908 14656 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector PRExpress, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14656)
AUTHORS Akmammedov A, Geigges M, Paro R.
TITLE Single vector non-leaky gene expression system for Drosophila
melanogaster
JOURNAL Sci Rep 7 (1), 6899 (2017)
PUBMED 28761084
REFERENCE 2 (bases 1 to 14656)
AUTHORS Akmammedov A, Geigges M, Paro R.
TITLE Direct Submission
JOURNAL Submitted (04-MAY-2017) Biosystem Science and Engineering, ETH
Zurich, Mattenstrasse 26, Basel 4058, Switzerland
REFERENCE 3 (bases 1 to 14656)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 14656)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep";
date: "2017"; volume: "7"; issue: "1"; pages: "6899"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(04-MAY-2017) Biosystem Science and Engineering, ETH Zurich,
Mattenstrasse 26, Basel 4058, Switzerland"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..14656
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(3..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 600..616
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 623..641
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
gene 666..4802
/label=mini-white
/note="This modified version of the white gene lacks part
of the first intron."
protein_bind 4842..5227
/label=gypsy insulator
/note="chromatin insulator from Drosophila"
polyA_signal complement(5235..5661)
/label=hsp70 poly(A)
/note="polyadenlyation signal from the Drosophila
melanogaster hsp70Ab gene"
CDS complement(5680..8724)
/codon_start=1
/label=lacZ
/note="beta-galactosidase"
/translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS
LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP
FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF
DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR
FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV
TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP
LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL
YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA
NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS
LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP
WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL
FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ
PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ
SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA
ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP
ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR
CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD
SWSPSVSAELQLSAGRYHYQLVWCQK"
promoter complement(8846..9294)
/label=hsp70 promoter
/note="Drosophila melanogaster hsp70Bb promoter"
misc_feature complement(9307..11747)
/label=poly-PRE
/note="poly-PRE"
protein_bind 11755..12140
/label=gypsy insulator
/note="chromatin insulator from Drosophila"
protein_bind 12180..12249
/label=attB
/note="attB site for the phi-C31 integrase (Groth et al.,
2000)"
promoter complement(12468..12486)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(12507..12523)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 12531..12547
/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(12555..12585)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(12600..12621)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(12909..13497)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(13671..14528)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(14529..14633)
/label=AmpR promoter
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