Basic Vector Information
- Vector Name:
- PRFP
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3469 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Zhang J.
PRFP 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 ℃.
PRFP vector Sequence
LOCUS 40924_36923 3469 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector PRFP, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3469)
AUTHORS Zhang J.
TITLE Direct Submission
JOURNAL Submitted (17-AUG-2016) Research Institute of Pomology, Chinese
Academy of Agricultural Sciences, Xingcheng, Liaoning 125100, China
REFERENCE 2 (bases 1 to 3469)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 3469)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
(17-AUG-2016) Research Institute of Pomology, Chinese Academy of
Agricultural Sciences, Xingcheng, Liaoning 125100, China"
COMMENT SGRef: number: 2; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..3469
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 379..395
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 415..433
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 491..1177
/codon_start=1
/label=TagRFP
/note="monomeric derivative of red fluorescent protein from
Entacmaea quadricolor (Merzlyak et al., 2007)"
/translation="ELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKVVE
GGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTATQ
DTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSDMALKLV
GGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVARYCDL
PSKLGHK"
primer_bind complement(1248..1264)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(1272..1288)
/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(1296..1326)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(1341..1362)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(1650..2238)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2412..3269)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQAQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFLADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAAIGAS
LIKHW"
promoter complement(3270..3374)
/label=AmpR promoter
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