Basic Vector Information
- Vector Name:
- pOtta_sfGFP__pOsym_mKate2
- Antibiotic Resistance:
- Kanamycin
- Length:
- 5113 bp
- Type:
- Cloning vector
- Replication origin:
- pSC101 ori
- Source/Author:
- Groseclose TM, Hersey AN, Huang BD, Wilson CJ.
pOtta_sfGFP__pOsym_mKate2 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 ℃.
pOtta_sfGFP__pOsym_mKate2 vector Sequence
LOCUS 62056_18310 5113 bp DNA circular SYN 16-AUG-2021
DEFINITION Cloning vector pOtta_sfGFP__pOsym_mKate2, complete sequence.
ACCESSION MZ198787
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5113)
AUTHORS Groseclose TM, Hersey AN, Huang BD, Wilson CJ.
TITLE Direct Submission
JOURNAL
REFERENCE 2 (bases 1 to 5113)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
(13-MAY-2021) Chemical "
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..5113
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator complement(400..494)
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(528..1319)
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
regulatory 1506..1534
/label=trc promoter
/note="trc promoter"
/regulatory_class="promoter"
regulatory 1506..1511
/label=trc promoter -35 hexamer
/note="trc promoter -35 hexamer"
/regulatory_class="promoter"
regulatory 1512..1528
/label=operator Otta
/note="operator Otta"
/regulatory_class="other"
regulatory 1529..1534
/label=trc promoter -10 hexamer
/note="trc promoter -10 hexamer"
/regulatory_class="promoter"
regulatory 1553..1564
/label=strong bacterial RBS
/note="strong bacterial RBS"
/regulatory_class="ribosome_binding_site"
CDS 1571..2284
/label=superfolder GFP
/note="GFP variant that folds robustly even when fused to
poorly folded proteins (Nager et al., 2011)"
terminator 2299..2385
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(2926..3873)
/label=Rep101
/note="RepA protein needed for replication with the pSC101
origin"
rep_origin complement(3921..4143)
/direction=LEFT
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
protein_bind 4256..4275
/label=lac operator (symmetric)
/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). The
symmetric lac operator was optimized for tight binding of
lac repressor."
regulatory 4275..4280
/label=trc promoter -10 hexamer
/note="trc promoter -10 hexamer"
/regulatory_class="promoter"
regulatory 4299..4310
/label=strong bacterial RBS
/note="strong bacterial RBS"
/regulatory_class="ribosome_binding_site"
CDS 4317..5012
/label=mKate2
/note="monomeric far-red fluorescent protein (Shcherbo et
al., 2009)"
terminator complement(5030..5073)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
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