Basic Vector Information
- Vector Name:
- pPA1
- Antibiotic Resistance:
- Streptomycin
- Length:
- 7140 bp
- Type:
- Cloning vector
- Replication origin:
- pBBR1 oriV
- Source/Author:
- Ghosh IN, Landick R.
pPA1 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 ℃.
pPA1 vector Sequence
LOCUS 40924_34046 7140 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pPA1, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7140)
AUTHORS Ghosh IN, Landick R.
TITLE OptSSeq: High-throughput sequencing readout of growth enrichment
defines optimal gene expression elements for homoethanologenesis
JOURNAL ACS Synth Biol (2016) In press
PUBMED 27404024
REFERENCE 2 (bases 1 to 7140)
AUTHORS Ghosh IN, Landick RC.
TITLE Direct Submission
JOURNAL Submitted (15-JUL-2016) DOE Great Lakes Bioenergy Research Center,
University of Wisconsin-Madison, 1552 University Avenue, Madison, WI
53726, United States
REFERENCE 3 (bases 1 to 7140)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7140)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
Biol (2016) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(15-JUL-2016) DOE Great Lakes Bioenergy Research Center, University
of Wisconsin-Madison, 1552 University Avenue, Madison, WI 53726,
United States"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7140
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(161..820)
/label=pBBR1 Rep
/note="replication protein for the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica"
rep_origin complement(821..1590)
/direction=LEFT
/label=pBBR1 oriV
/note="replication origin of the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica; requires the pBBR1
Rep protein for replication"
promoter 2968..3059
/label=AmpR promoter
CDS 3060..3848
/label=SmR
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
regulatory 4048..4095
/label=B. subtilis glnA terminator
/note="B. subtilis glnA terminator"
/regulatory_class="terminator"
protein_bind complement(4107..4126)
/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 4154..4182
/label=Promoter#27
/note="Promoter#27"
/regulatory_class="promoter"
regulatory 4236..4244
/label=pdc RBS#3
/note="pdc RBS#3"
/regulatory_class="ribosome_binding_site"
CDS 4250..5953
/gene="pdc"
/label=pdc
/note="Pyruvate decarboxylase from Zymomonas mobilis subsp.
mobilis (strain ATCC 31821 / ZM4 / CP4). Accession#:
P06672"
regulatory 5972..5980
/label=adhA RBS#2
/note="adhA RBS#2"
/regulatory_class="ribosome_binding_site"
CDS 5986..6996
/gene="adhA"
/label=adhA
/note="Alcohol dehydrogenase 1 from Zymomonas mobilis
subsp. mobilis (strain ATCC 31821 / ZM4 / CP4). Accession#:
P20368"
regulatory 7044..7095
/label=Phage P22 terminator
/note="Phage P22 terminator"
/regulatory_class="terminator"
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