Tol2-elavl3-H2B-GCaMP6s vector (V006782)

Price Information

Cat No. Plasmid Name Availability Add to cart
V006782 Tol2-elavl3-H2B-GCaMP6s In stock, 1 week for quality controls

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Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:
Tol2-elavl3-H2B-GCaMP6s
Antibiotic Resistance:
Ampicillin
Length:
14296 bp
Type:
Transposable element
Replication origin:
ori
Copy Number:
High Copy
Cloning Method:
Gibson Cloning
5' Primer:
ACTCCAGCAGGACCATGTGAT
3' Primer:
CATCGATGGTACAGTAAAACGACGG

Tol2-elavl3-H2B-GCaMP6s vector Vector Map

Tol2-elavl3-H2B-GCaMP6s14296 bp7001400210028003500420049005600630070007700840091009800105001120011900126001330014000F1ori-FH2BGCaMP6sSV40 poly(A) signalSK primerT3 promoterM13 revlac operatorlac promoterCAP binding siteL4440oriAmpRAmpR promoter

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 ℃.

Tol2-elavl3-H2B-GCaMP6s vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_48957       14296 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Expressed nuclear-localized GCaMP6-slow, a calcium indicator, in 
            neurons in zebrafish larva. The plasmid is in Tol2 transposon 
            backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14296)
  AUTHORS   Freeman J, Vladimirov N, Kawashima T, Mu Y, Sofroniew NJ, Bennett 
            DV, Rosen J, Yang CT, Looger LL, Ahrens MB
  TITLE     Mapping brain activity at scale with cluster computing.
  JOURNAL   Nat Methods. 2014 Jul 27. doi: 10.1038/nmeth.3041.
  PUBMED    25068736
REFERENCE   2  (bases 1 to 14296)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 14296)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2014 Jul 27. doi: 10.1038/nmeth.3041."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..14296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             9463..9840
                     /codon_start=1
                     /label=H2B
                     /note="human histone H2B"
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             9859..11208
                     /codon_start=1
                     /label=GCaMP6s
                     /note="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     polyA_signal    complement(11253..11387)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12059..12075)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12059..12075)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(12112..12130)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(12151..12167)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12151..12167)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12164..12186)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12175..12191
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(12199..12229)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(12244..12265)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(12382..12399)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12553..13141)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(13315..14172)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(14173..14277)
                     /label=AmpR promoter