Basic Vector Information
- Vector Name:
- pTimer
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3314 bp
- Type:
- Fluorescent Protein Genes & Plasmids
- Replication origin:
- ori
- Source/Author:
- Clontech
- Copy Number:
- High copy number
- 3' Primer:
- M13 rev
pTimer 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 ℃.
pTimer vector Sequence
LOCUS 40924_43363 3314 bp DNA circular SYN 01-JAN-1980 DEFINITION Vector for expressing the Fluorescent Timer in bacteria. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3314) AUTHORS Clontech TITLE Direct Submission REFERENCE 2 (bases 1 to 3314) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" COMMENT Translation from the lacZ start codon produces a fusion protein in E. coli. FEATURES Location/Qualifiers source 1..3314 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind 107..128 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 143..173 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 181..197 /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)." primer_bind 205..221 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" CDS 289..966 /codon_start=1 /label=Timer /note="green-to-red fluorescent timer derivative of DsRed (Terskikh et al., 2000)" /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG VATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERTEGRH HLFL" promoter 1409..1513 /label=AmpR promoter CDS 1514..2371 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 2545..3133 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
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