Basic Vector Information
- Vector Name:
- ArcLight-A242
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7143 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Selection Marker:
- Hygromycin
- Copy Number:
- High Copy
- Promoter:
- sCMV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- GACGTAAATGGGCGGTAGGCGTG
- 3' Primer:
- TTAAAAAACCTCCCACACCTC
ArcLight-A242 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 ℃.
ArcLight-A242 vector Sequence
LOCUS 40924_290 7143 bp DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7143) AUTHORS Jin L, Han Z, Platisa J, Wooltorton JR, Cohen LB, Pieribone VA TITLE Single action potentials and subthreshold electrical events imaged in neurons with a fluorescent protein voltage probe. JOURNAL Neuron. 2012 Sep 6;75(5):779-85. PUBMED 22958819 REFERENCE 2 (bases 1 to 7143) TITLE Direct Submission REFERENCE 3 (bases 1 to 7143) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Neuron."; date: "2012-09-6"; volume: "75(5)"; pages: "779-85" COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..7143 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 35..53 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" CDS 799..1512 /codon_start=1 /label=superecliptic pHluorin /note="pH-sensitive mutant of green fluorescent protein (Ng et al., 2002)" /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKA NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLE FVTADGITHGMDELYK" promoter complement(1523..1540) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal complement(1545..1679) /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" promoter 1789..2118 /label=SV40 promoter /note="SV40 enhancer and early promoter" CDS 2167..3189 /codon_start=1 /label=HygR /note="aminoglycoside phosphotransferase from E. coli" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" polyA_signal 3322..3455 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3481..3497) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(3505..3521) /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(3529..3559) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(3574..3595) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3712..3729) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3883..4471) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4645..5502) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(5503..5607) /label=AmpR promoter rep_origin complement(5633..6088) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 6165..7143 /label=CMV IE94 promoter /note="enhancer/promoter region of simian cytomegalovirus major immediate early transcription unit IE94"
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