Tol2-elavl3-H2B-GCaMP6s vector (V006782)

Basic Vector Information

      • Vector Name:
      • Tol2-elavl3-H2B-GCaMP6s
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 14296 bp
      • Type:
      • Transposable element
      • 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-FH2BKozak sequenceSV40 poly(A) signalSK primerT3M13 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       Exported               14296 bp ds-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    Tol2-elavl3-H2B-GCaMP6s
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)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2014 Jul 27. doi: 10.1038/nmeth.3041."
FEATURES             Location/Qualifiers
     source          1..14296
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             9463..9840
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      9853..9862
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             9859..11211
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      9955..9964
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     10245..10266
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(10372..10393)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(10633..10652)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    11253..11387
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11277..11296)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     11331..11350
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     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"
     primer_bind     complement(12112..12132)
                     /label=T3
                     /note="T3 promoter, forward primer"
     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    12244..12265
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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"
     primer_bind     complement(12633..12652)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(13312..14172)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     13935..13954
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14173..14277)
                     /gene="bla"
                     /label=AmpR promoter

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