Basic Vector Information
- Vector Name:
- pOT2-3GLA
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 3293 bp
- Type:
- Cloning and transformation vector
- Replication origin:
- ori
- Source/Author:
- Haussmann IU, Brauer U, Ustaoglu P, Hemani Y, Soller M.
- Promoter:
- tet
pOT2-3GLA 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 ℃.
pOT2-3GLA vector Sequence
LOCUS 40924_33942 3293 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning and transformation vector pOT2-3GLA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3293)
AUTHORS Haussmann IU, Brauer U, Ustaoglu P, Hemani Y, Soller M.
TITLE Plasmid based gap-repair recombineering for Drosophila transgenesis
and the analysis of alternative pre-mRNA processing
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 3293)
AUTHORS Haussmann IU, Brauer U, Ustaoglu P, Hemani Y, Soller M.
TITLE Direct Submission
JOURNAL Submitted (15-OCT-2014) School of Bioscience, University of
Birmingham, Edgbaston, Birmingham, West Midlands B15 2TT, United
Kingdom
REFERENCE 3 (bases 1 to 3293)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3293)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(15-OCT-2014) School of Bioscience, University of Birmingham,
Edgbaston, Birmingham, West Midlands B15 2TT, United Kingdom"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Assembly Method :: DNASTAR Lasergene SeqBuilder v. 7.2.1
Sequencing Technology :: not fully sequenced, assembled from pieces,
vector fully functional
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..3293
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind complement(188..257)
/label=attB
/note="attB site for the phi-C31 integrase (Groth et al.,
2000)"
promoter complement(387..405)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
CDS complement(473..1129)
/label=CmR
/note="chloramphenicol acetyltransferase"
promoter complement(1166..1194)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
rep_origin complement(1256..1844)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 1948..1966
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 2004..2095
/label=multiple cloning site
/note="multiple cloning site"
protein_bind 2100..2133
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
misc_feature 2138..2374
/label=3xP3-hsp70
/note="3xP3-hsp70"
regulatory 2369..2378
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 2390..3100
/label=EGFP
/note="enhanced GFP"
misc_feature 3137..3236
/note="ewg PA1; polyadenylation site 1 of the Drosophila
erect wing gene"
misc_recomb 3248..3281
/label=loxP site for cre mediated recombination
/note="loxP site for cre mediated recombination"
protein_bind complement(3248..3281)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
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