Basic Vector Information
- Vector Name:
- pOpt_Clover_Paro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6671 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Lauersen KJ, Mussgnug JH, Kruse O.
pOpt_Clover_Paro 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 ℃.
pOpt_Clover_Paro vector Sequence
LOCUS 40924_33822 6671 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pOpt_Clover_Paro, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6671)
AUTHORS Lauersen KJ, Mussgnug JH, Kruse O.
TITLE Efficient targeted expression of nuclear transgenes in Chlamydomonas
reinhardtii with a novel, modular vector toolkit
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6671)
AUTHORS Lauersen KJ, Mussgnug JH, Kruse O.
TITLE Direct Submission
JOURNAL Submitted (25-JUN-2014) Algae Biotechnology and Bioenergy,
University of Bielefeld Center for Biotechnology, Universitaetsstr
27, Bielefeld, NRW 33615, Germany
REFERENCE 3 (bases 1 to 6671)
AUTHORS Lauersen KJ, Mussgnug JH, Kruse O.
TITLE Direct Submission
JOURNAL Submitted (18-MAY-2016) Algae Biotechnology and Bioenergy,
University of Bielefeld Center for Biotechnology, Universitaetsstr
27, Bielefeld, NRW 33615, Germany
REFERENCE 4 (bases 1 to 6671)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 6671)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(25-JUN-2014) Algae Biotechnology and Bioenergy, University of
Bielefeld Center for Biotechnology, Universitaetsstr 27, Bielefeld,
NRW 33615, Germany"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(18-MAY-2016) Algae Biotechnology and Bioenergy, University of
Bielefeld Center for Biotechnology, Universitaetsstr 27, Bielefeld,
NRW 33615, Germany"
COMMENT SGRef: number: 4; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
On May 18, 2016 this sequence version replaced KM061062.1.
FEATURES Location/Qualifiers
source 1..6671
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 489..505
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 515..533
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1247..1258
/codon_start=1
/label=Factor Xa site
/note="Factor Xa recognition and cleavage site"
/translation="IEGR"
CDS 2269..2280
/codon_start=1
/label=Factor Xa site
/note="Factor Xa recognition and cleavage site"
/translation="IEGR"
CDS 2293..2316
/codon_start=1
/label=Strep-Tag II
/note="peptide that binds Strep-Tactin(R), an engineered
form
of streptavidin"
/translation="WSHPQFEK"
promoter complement(4371..4389)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(4410..4426)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4434..4450)
/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(4458..4488)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4503..4524)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4812..5400)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5574..6431)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(6432..6536)
/label=AmpR promoter
rep_origin complement(join(6563..6671,1..347))
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
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