pAAV-hsyn-GRAB_5-HT1.0 vector (V012731)

Basic Vector Information

Expresses the 5-HT sensor GRAB_5-HT1.0 in neurons, could be used as a genetically encoded sensor for measuring serotonin dynamics

      • Vector Name:
      • pAAV-hsyn-GRAB_5-HT1.0
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 6530 bp
      • Type:
      • Viral Expression & Packaging Vectors
      • Source/Author:
      • Yulong Li
      • Copy Number:
      • High copy number
      • Promoter:
      • human synapsin
      • Fusion Tag:
      • cpEGFP
      • Expression Method:
      • Constiutive, Stable

pAAV-hsyn-GRAB_5-HT1.0 vector Vector Map

pAAV-hsyn-GRAB_5-HT1.06530 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300AAV2 ITR (alternate)synapsinIg-kappa leaderattB1Human HTR2C(2-247)VC155Human HTR2C(304-458)WPREhGH poly(A) signalAAV2 ITRf1 oriAmpR promoterAmpRori

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 ℃.

pAAV-hsyn-GRAB_5-HT1.0 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       pAAV-hsyn-GRAB_5        6530 bp DNA     circular SYN 09-APR-2021
DEFINITION  Expresses the 5-HT sensor GRAB_5-HT1.0 in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-hsyn-GRAB_5-HT1.0.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6530)
  TITLE     Yulong Li 5-HT sensor and DA sensor plasmids
REFERENCE   2  (bases 1 to 6530)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6530)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6530
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="AAV2 ITR (alternate)"
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        145..586
                     /note="synapsin"
                     /note="human synapsin promoter is a neuron-specific
                     promoter"
     regulatory      581..590
                     /label=Kozak sequence
                     /note="Kozak sequence"
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             587..649
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa
                     light chain"
                     /label=leader sequence from mouse immunoglobulin kappa
                     /note="Ig-kappa leader"
                     /translation="METDTLLLWVLLLWVPGSTGD"
     protein_bind    650..674
                     /gene="mutant version of attB"
                     /label=BP Clonase(TM) binding site
                     /bound_moiety="BP Clonase(TM)"
                     /note="attB1"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             683..1420
                     /codon_start=1
                     /label=Human HTR2C(2-247)
                     /note="Human HTR2C(2-247)"
                     /note="G-protein coupled receptor for 5-hydroxytryptamine 
                     (serotonin). Also functions as a receptor for various drugs
                     and psychoactive substances, including ergot alkaloid 
                     derivatives, 1-2,5,-dimethoxy-4-iodophenyl-2-aminopropane 
                     (DOI) and lysergic acid diethylamide (LSD). Ligand binding 
                     causes a conformation change that triggers signaling via 
                     guanine nucleotide-binding proteins (G proteins) and 
                     modulates the activity of down-stream effectors. 
                     Beta-arrestin family members inhibit signaling via G 
                     proteins and mediate activation of alternative signaling 
                     pathways. Signaling activates a 
                     phosphatidylinositol-calcium second messenger system that 
                     modulates the activity of phosphatidylinositol 3-kinase and
                     down-stream signaling cascades and promotes the release of 
                     Ca2+ ions from intracellular stores. Regulates neuronal 
                     activity via the activation of short transient receptor 
                     potential calcium channels in the brain, and thereby 
                     modulates the activation of pro-opiomelacortin neurons and 
                     the release of CRH that then regulates the release of 
                     corticosterone. Plays a role in the regulation of appetite 
                     and eating behavior, responses to anxiogenic stimuli and 
                     stress. Plays a role in insulin sensitivity and glucose 
                     homeostasis."
                     /translation="VNLRNAVHSFLVHLIGLLVWQCDISVSPVAAIVTDIFNTSDGGRF
                     KFPDGVQNWPALSIVIIIIMTIGGNILVIMAVSMEKKLHNATNYFLMSLAIADMLVGLL
                     VMPLSLLAILYDYVWPLPRYLCPVWISLDVLFSTASIMHLCAISLDRYVAIRNPIEHSR
                     FNSRTKAIMKIAIVWAISIGVSVPIPVIGLRDEEKVFVNNTTCVLNDPNFVLIGSFVAF
                     FIPLTIMVITYCLTIYVLRRQALM"
     CDS             1451..1702
                     /label=VC155
                     /note="C-terminal fragment of mVenus for use in bimolecular
                     fluorescence complementation (BiFC) (Kodama and Hu, 2010)"
     CDS             2177..2644
                     /codon_start=1
                     /label=Human HTR2C(304-458)
                     /note="Human HTR2C(304-458)"
                     /translation="NNERKASKVLGIVFFVFLIMWCPFFITNILSVLCEKSCNQKLMEK
                     LLNVFVWIGYVCSGINPLVYTLFNKIYRRAFSNYLRCNYKVEKKPPVRQIPRVAATALS
                     GRELNVNIYRHTNEPVIEKASDNEPGIEMQVENLELPVNPSSVVSERISSV"
     misc_feature    2657..3245
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    3277..3753
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   3793..3933
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4008..4463
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4745..4849
                     /label=AmpR promoter
     CDS             4850..5707
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5881..6469
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

This page is informational only.