Basic Vector Information
- Vector Name:
- pNK076
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6737 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Kandul NP, Guo M, Hay BA.
- Promoter:
- Ac5
pNK076 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 ℃.
pNK076 vector Sequence
LOCUS 40924_33322 6737 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector pNK076, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6737) AUTHORS Kandul NP, Guo M, Hay BA. TITLE A positive readout single transcript reporter for site-specific mRNA cleavage JOURNAL PeerJ (2017) In press REFERENCE 2 (bases 1 to 6737) AUTHORS Kandul NP, Guo M, Hay BA. TITLE Direct Submission JOURNAL Submitted (02-JAN-2017) Biology and Biological Engineering, California Institute of Technology, 1200 E. California Blvd., Pasadena, CA 91125, USA REFERENCE 3 (bases 1 to 6737) TITLE Direct Submission REFERENCE 4 (bases 1 to 6737) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PeerJ (2017) In press" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (02-JAN-2017) Biology and Biological Engineering, California Institute of Technology, 1200 E. California Blvd., Pasadena, CA 91125, 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..6737 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin 4..460 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 601..617 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 627..645 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter 699..3208 /label=Ac5 promoter /note="Drosophila melanogaster actin 5C promoter" CDS 3236..4168 /codon_start=1 /label=Rluc /note="luciferase from the anthozoan coelenterate Renilla reniformis (sea pansy)" /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ" promoter complement(4549..4567) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(4588..4604) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(4612..4628) /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(4636..4666) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(4681..4702) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4990..5578) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(5752..6609) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(6610..6714) /label=AmpR promoter
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