Basic Vector Information
- Vector Name:
- mTol2-B-P-Gal4VP16-Cre-CG3
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9078 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Otsuna H, Scoresby AN, Gaynes B, Tong ZZ, Kwan KK, Hutcheson DA, Chien CB, Dorsky R.
- Promoter:
- cmlc2
mTol2-B-P-Gal4VP16-Cre-CG3 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 ℃.
mTol2-B-P-Gal4VP16-Cre-CG3 vector Sequence
LOCUS 40924_2144 9078 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector mTol2-B-P-Gal4VP16-Cre-CG3, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9078)
AUTHORS Otsuna H, Scoresby AN, Gaynes B, Tong ZZ, Kwan KK, Hutcheson DA,
Chien CB, Dorsky R.
TITLE Large Scale Zebrafish Gal4 Enhancer Trap Screen for Neural Specific
Expression Patterns
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 9078)
AUTHORS Otsuna H, Scoresby AN, Gaynes B, Tong ZZ, Kwan KK, Hutcheson DA,
Chien CB, Dorsky R.
TITLE Direct Submission
JOURNAL Submitted (19-MAR-2013) Neurobiology and Anatomy, University of
Utah, 20 North 1900 East, Salt Lake City, UT 84132, USA
REFERENCE 3 (bases 1 to 9078)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9078)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(19-MAR-2013) Neurobiology and Anatomy, University of Utah, 20 North
1900 East, Salt Lake City, UT 84132, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..9078
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(1..18)
/label=T7 primer
/note="T7 primer"
primer_bind complement(24..41)
/label=M13F
/note="M13F"
primer_bind complement(24..40)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 112..180
/label=LacZ alpha
/note="LacZ alpha"
rep_origin 181..636
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 714..818
/label=AmpR promoter
CDS 819..1676
/label=AmpR
/note="beta-lactamase"
rep_origin 1850..2438
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2726..2747
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2762..2792
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2800..2816
/label=lac operator
/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)."
primer_bind 2824..2840
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 2858..2876
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
repeat_region 2927..3085
/label=miniTol2 3' end
/note="miniTol2 3' end"
repeat_region 2927..2943
/label=Tol2 3' TIR
/note="Tol2 3' TIR"
primer_bind 3108..3129
/label=M13R
/note="M13R"
primer_bind 3108..3124
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3144..3164
/label=attB4
/note="core recombination site for the Gateway(R) BP
reaction"
regulatory 3193..3203
/label=adenovirus E1B TATA
/note="adenovirus E1B TATA"
/regulatory_class="promoter"
5'UTR 3211..3242
/label=carp beta-actin trx start
/note="carp beta-actin trx start"
protein_bind 3249..3282
/label=FRT (minimal)
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
protein_bind 3283..3307
/label=attB1
/note="recombination site for the Gateway(R) BP reaction"
CDS 3313..3753
/label=GAL4 DNA binding domain
/note="DNA binding domain of the GAL4 transcriptional
activator"
misc_feature 3790..3963
/label=VP16 activation domain
/note="VP16 activation domain"
protein_bind complement(3968..3992)
/label=attB2
/note="recombination site for the Gateway(R) BP reaction"
promoter complement(3993..4010)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 4015..4212
/label=SV40 late polyA (from pCS2+)(1)
/note="SV40 late polyA (from pCS2+)(1)"
polyA_signal 4015..4149
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
misc_feature complement(4261..4699)
/label=yeast ded1/HIS3 protein fragment X03245.1
/note="yeast ded1/HIS3 protein fragment X03245.1"
intron 4706..4777
/note="SV40 small T antigen intron"
intron 4782..4847
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 4977..4997
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
polyA_signal 5422..5556
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
misc_recomb 5598..5631
/label=FRT
/note="FRT"
misc_feature 5598..5631
/label=FRT
/note="FRT"
protein_bind 5598..5631
/label=FRT (minimal)
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
CDS 5651..5671
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5669..6697
/label=Cre
/note="site-specific recombinase"
protein_bind complement(6712..6732)
/label=attB3
/note="core recombination site for the Gateway(R) BP
reaction"
primer_bind complement(6740..6756)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
polyA_signal complement(6772..6906)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
misc_feature complement(7028..7160)
/label=SV40 late polyA from pCS2+(2)
/note="SV40 late polyA from pCS2+(2)"
promoter 7165..7182
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS complement(7211..7927)
/label=EGFP
/note="enhanced GFP"
promoter complement(7928..8826)
/label=cmlc2
/note="Cardiac myosin light chain 2 promoter"
repeat_region 8850..9064
/label=miniTol2 5' end
/note="miniTol2 5' end"
repeat_region 9046..9064
/label=Tol2 5' TIR
/note="Tol2 5' TIR"
primer_bind complement(9074..9078)
/label=T7 primer(1)
/note="T7 primer(1)"
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