Basic Vector Information
- Vector Name:
- YCplac22 5-1.2-FLuciferase
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7428 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Belmont BJ, Niles JC.
- Promoter:
- TRP1
YCplac22 5-1.2-FLuciferase 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 ℃.
YCplac22 5-1.2-FLuciferase vector Sequence
LOCUS 40924_49467 7428 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector YCplac22 5-1.2-FLuciferase, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7428) AUTHORS Belmont BJ, Niles JC. TITLE Inducible Control of Subcellular RNA Localization Using a Synthetic Protein-RNA Aptamer Interaction JOURNAL PLoS ONE 7 (10), E46868 (2012) PUBMED 23056498 REFERENCE 2 (bases 1 to 7428) AUTHORS Belmont BJ, Niles JC. TITLE Direct Submission JOURNAL Submitted (12-SEP-2012) Biological Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave, Cambridge, MA 02139, USA REFERENCE 3 (bases 1 to 7428) TITLE Direct Submission REFERENCE 4 (bases 1 to 7428) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; date: "2012"; volume: "7"; issue: "10"; pages: "E46868" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (12-SEP-2012) Biological Engineering, Massachusetts Institute of Technology, 77 Massachusetts Ave, Cambridge, MA 02139, USA" COMMENT SGRef: number: 3; type: "Journal Article" COMMENT ##Assembly-Data-START## Sequencing Technology :: Sanger dideoxy sequencing ##Assembly-Data-END## FEATURES Location/Qualifiers source 1..7428 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 17..33 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS 678..2327 /codon_start=1 /label=luciferase /note="firefly luciferase" /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL" primer_bind complement(2632..2648) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" misc_feature complement(3407..4569) /label=CEN/ARS /note="S. cerevisiae CEN4 centromere fused to the autonomously replicating sequence ARS1/ARS416" promoter complement(5083..5184) /label=TRP1 promoter promoter 5290..5394 /label=AmpR promoter CDS 5395..6252 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 6426..7014 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 7302..7323 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 7338..7368 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7376..7392 /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 7400..7416 /label=M13 rev /note="common sequencing primer, one of multiple similar variants"
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