Basic Vector Information
DIO-SynaptoTag 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 ℃.
DIO-SynaptoTag vector Sequence
LOCUS 40924_670 7171 bp DNA circular SYN 17-DEC-2018 DEFINITION Cloning vector DIO-SynaptoTag, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7171) AUTHORS Zhou M, Liu Z, Melin MD, Ng YH, Xu W, Sudhof TC. TITLE A central amygdala to zona incerta projection is required for acquisition and remote recall of conditioned fear memory JOURNAL Nat. Neurosci. 21 (11), 1515-1519 (2018) PUBMED 30349111 REFERENCE 2 (bases 1 to 7171) AUTHORS Zhou M. TITLE Direct Submission JOURNAL Submitted (08-SEP-2018) Molecular and Cellular Physiology, Stanford University, 265 Campus Drive, Room G1065, Stanford, CA 94305, USA REFERENCE 3 (bases 1 to 7171) TITLE Direct Submission REFERENCE 4 (bases 1 to 7171) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Neurosci."; date: "2018"; volume: "21"; issue: "11"; pages: "1515-1519" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (08-SEP-2018) Molecular and Cellular Physiology, Stanford University, 265 Campus Drive, Room G1065, Stanford, CA 94305, USA" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..7171 /mol_type="other DNA" /organism="synthetic DNA construct" repeat_region 1..141 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" promoter 177..624 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" protein_bind 633..666 /label=lox2272 /note="Cre-mediated recombination occurs in the 8-bp core sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind 729..762 /label=loxP /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." CDS complement(1129..1845) /label=EGFP /note="enhanced GFP" misc_feature complement(1862..2419) /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS complement(2428..3132) /label=mCherry /note="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" regulatory 3138..3147 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" protein_bind complement(3148..3181) /label=lox2272 /note="Cre-mediated recombination occurs in the 8-bp core sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." protein_bind complement(3252..3285) /label=loxP /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." misc_feature 3298..3886 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" polyA_signal 3918..4394 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" repeat_region 4434..4574 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" rep_origin 4649..5104 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 5386..5490 /label=AmpR promoter CDS 5491..6348 /label=AmpR /note="beta-lactamase" rep_origin 6522..7110 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
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