Basic Vector Information
- Vector Name:
- G-CaMP3
- Antibiotic Resistance:
- Kanamycin
- Length:
- 5311 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- EGFP-N
G-CaMP3 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 ℃.
G-CaMP3 vector Sequence
LOCUS 40924_934 5311 bp DNA circular SYN 13-MAY-2021
DEFINITION a single-wavelength GCaMP2-based genetically encoded calcium
indicator.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5311)
AUTHORS Tian L, Hires SA, Mao T, Huber D, Chiappe ME, Chalasani SH, Petreanu
L, Akerboom J, McKinney SA, Schreiter ER, Bargmann CI, Jayaraman V,
Svoboda K, Looger LL.
TITLE Imaging neural activity in worms, flies and mice with improved GCaMP
calcium indicators.
JOURNAL Nat Methods. 2009 Dec;6(12):875-81.
PUBMED 19898485
REFERENCE 2 (bases 1 to 5311)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 5311)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Methods."; date: "2009-12"; volume: "6(12)"; pages: "875-81"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5311
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..204
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 261..1610
/codon_start=1
/label=GCaMP6f
/note="improved fluorescent protein-based calcium sensor
(Chen et al., 2013)"
/translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
LKGIDFKEDGNILGHKLEYNTRDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGN
GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
polyA_signal 1735..1856
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin complement(1863..2318)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2345..2449
/label=AmpR promoter
promoter 2451..2808
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 2843..3634
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3825..3844)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3869..3916
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4245..4833
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
enhancer 5008..5311
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
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