Basic Vector Information
- Vector Name:
- pNHEJ-RPG
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9062 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells
- Promoter:
- CAG
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pNHEJ-RPG 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 ℃.
pNHEJ-RPG vector Sequence
LOCUS 62056_17970 9062 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9062) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..9062 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 26..130 /label=AmpR promoter CDS 131..988 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 1162..1750 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 2038..2059 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 2074..2104 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2112..2128 /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 2136..2152 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 2173..2191 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 3141..3344 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" CDS 3384..4061 /codon_start=1 /label=DsRed1 /note="wild-type DsRed" /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH HLFL" polyA_signal 4133..4188 /label=beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" intron 5220..6236 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" CDS 6990..7703 /codon_start=1 /label=EGFP /note="enhanced GFP" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIDDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" polyA_signal 7824..7945 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" promoter complement(8424..8442) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(8449..8465) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 8607..9062 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
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