Basic Vector Information
- Vector Name:
- ploxP_cPeUMPS
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6127 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Kasai Y, Tsukahara T, Ikeda F, Ide Y, Harayama S.
ploxP_cPeUMPS 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 ℃.
ploxP_cPeUMPS vector Sequence
LOCUS 40924_28611 6127 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector ploxP_cPeUMPS DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6127)
AUTHORS Kasai Y, Tsukahara T, Ikeda F, Ide Y, Harayama S.
TITLE Improving triacylglycerol production in the unicellular green alga
Pseudochoricystis ellipsoidea by metabolic engineering using
iterative self-cloning approaches
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6127)
AUTHORS Kasai Y, Harayama S.
TITLE Direct Submission
JOURNAL Submitted (27-OCT-2017) Contact:Yuki Kasai Chuo University, Research
and Development Initiative; 1-13-27 Kasuga, Bunkyo-ku, Tokyo
112-8551, Japan
REFERENCE 3 (bases 1 to 6127)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6127)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(27-OCT-2017) Contact:Yuki Kasai Chuo University, Research and
Development Initiative; 1-13-27 Kasuga, Bunkyo-ku, Tokyo 112-8551,
Japan"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6127
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(3..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 600..616
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 626..644
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 670..686
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 707..740
/label=loxP
/note="loxP"
protein_bind 707..740
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
regulatory 741..1530
/note="includes promoter region of Pseudochoricystis
ellipsoidea nuclear gene RBCS for ribulose bisphosphate
carboxylase/oxygenase small subunit"
/regulatory_class="promoter"
misc_feature 1531..1815
/note="include intron 1 of Pseudochoricystis ellipsoidea
nuclear gene RBCS for ribulose bisphosphate
carboxylase/oxygenase small subunit"
CDS 1816..3318
/codon_start=1
/gene="UMPS"
/product="uridine monophosphate synthase"
/label=UMPS
/protein_id="BBB04236.1"
/translation="MATSTPSVATAAELKPKEGPITSQEFEELVLRLHEIEAVKFGNFK
LKSGLMSPIYIDLRVIVSYPDVLRRVAEVMWHQVSGAAFDVMCGVPYTALPIATCMSLL
HGTPMLMRRKEVKEYGTKKAIEGAFKKGQTCLIVEDLVTSGASVMETVEPLEVEGLKVT
DVVVLIDREQGGAARMASNGLRLHSAFTLSFIIKTLQAHGLVSAKVADSVAAFIAANQT
FTPSAAAPTPSPAAPAQPKRLPFEERASLCQNAAGRKLLELMARKRTNLAVAADVATVE
EMLRIADAAGPHIAVFKTHVDIFDKWDDGIATQLRHLADKHEFLIFEDRKFADIGNTVV
SQYGGGIYKIADWSDITNAHLVPGPGIIDGLRKVGQEKGRGLLLLAEMSSKGALATGAY
TEKVAEAAAANQDFVMGFICQSPAKWATPVPPGLVHMTPGVQLASGSDALGQQYNTPAS
VIGQGGSDVIIVGRGIIKAADPAAAAAQYREAGWAAYEATLA"
gene 1816..3318
/gene="UMPS"
/label=UMPS
regulatory 3319..3850
/note="includes terminator region of Pseudochoricystis
ellipsoidea nuclear gene RBCS for ribulose bisphosphate
carboxylase/oxygenase small subunit"
/regulatory_class="terminator"
protein_bind complement(3851..3884)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
primer_bind complement(3886..3902)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(3939..3957)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(3978..3994)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4002..4018)
/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)."
promoter complement(4026..4056)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4071..4092)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4380..4968)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5142..5999)
/label=AmpR
/note="beta-lactamase"
promoter complement(6000..6104)
/label=AmpR promoter
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