Basic Vector Information
pCVD047 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 ℃.
pCVD047 vector Sequence
LOCUS 40924_49217 7140 bp DNA circular SYN 18-DEC-2018
DEFINITION Broad host range vector vector pCVD047, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7140)
AUTHORS Taton A, Unglaub F, Wright NE, Zeng WY, Paz-Yepez J, Brahamsha B,
Palenik B, Peterson TC, Haerizadeh F, Golden SS, Golden JW.
TITLE Broad-host-range vector system for synthetic biology and
biotechnology in cyanobacteria
JOURNAL Nucleic Acids Res. (2014) In press
PUBMED 25074377
REFERENCE 2 (bases 1 to 7140)
AUTHORS Taton A, Unglaub F, Wright N, Zeng WY, Paz Yepes J, Brahamsha B,
Palenik B, Peterson T, Haerizadeh F, Golden SS, Golden JW.
TITLE Direct Submission
JOURNAL Submitted (17-JUN-2014) Division of Biological Sciences, University
of California, San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA
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: "Nucleic
Acids Res. (2014) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(17-JUN-2014) Division of Biological Sciences, University of
California, San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7140
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(124..981)
/label=AmpR
/note="beta-lactamase"
promoter complement(982..1086)
/label=AmpR promoter
misc_feature 1112..1117
/label=ZraI
/note="ZraI"
misc_feature 1227..1232
/label=ZraI
/note="ZraI"
misc_feature 1230..1250
/note="G5C5; GC-adaptor for seamless assembly, flanking a
BHR replicons, neutral sites, or cyanobacterial replicons;
synthetic"
misc_feature 1251..1256
/label=XbaI
/note="XbaI"
regulatory 1288..1314
/note="trpA T; trpA terminator from Lorist6 INSD accession
X98450, one nt shorter than reference; Lorist6"
/regulatory_class="terminator"
CDS complement(1650..2498)
/label=RSF1010 RepC
/note="replication protein C of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
CDS complement(2488..3324)
/label=RSF1010 RepA
/note="replication protein A of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
CDS complement(3354..3560)
/codon_start=1
/product="repF"
/label=repF
/note="repF-RSF1010; replication gene; derived from
pRL1383a in INSD accession AF403426, a derivative of
RSF1010 INSD accession M28829"
/protein_id="AII99706.1"
/translation="MKDQKDKQTGDLLASPDAVRQARYAERMKAKGMRQRKFWLTDDEY
EALRECLEELRAAQGGGSDPASA"
CDS complement(3562..3774)
/codon_start=1
/product="repE"
/label=repE
/note="repE-RSF1010; replication gene; derived from
pRL1383a in INSD accession AF403426, a derivative of
RSF1010 INSD accession M28829"
/protein_id="AII99707.1"
/translation="MEYEKSASGSVYLIKSDKGYWLPGGFGYTSNKAEAGRFSVADMAS
LNLDGCTLSLFREDKPFGPGKFLGD"
regulatory complement(3804..3831)
/note="P4; derived from pRL1383a in INSD accession
AF403426, a derivative of RSF1010 INSD accession M28829;
the P4 promoter is under the auto-regulated control of the
F repressor, whose gene is located in the same operon,
providing transcription of E-F-repA-repC-operon"
/regulatory_class="promoter"
CDS complement(3838..4806)
/label=RSF1010 RepB
/note="replication protein B of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
CDS complement(4803..5216)
/codon_start=1
/product="mobB"
/label=mobB
/note="mobilization gene; derived from pRL1383a in INSD
accession AF403426, a derivative of RSF1010 INSD accession
M28829"
/protein_id="AII99704.1"
/translation="MNAIDRVKKSRGINELAEQIEPLAQSMATLADEARQVMSQTKQAS
EAQAAEWLKAQRQTGAAWVELAKELREVAAEVSSAAQSARSASRGWHWKLWLTVMLASM
MPTVVLLIASLLLLDLTPLTTEDGSIWLRLVAR"
misc_feature 5662..5667
/label=AgeI
/note="AgeI"
oriT complement(6045..6132)
/direction=LEFT
/label=RSF1010 oriT
/note="origin of transfer of the broad-host-range plasmid
RSF1010 (Scholz et al., 1989)"
regulatory complement(6141..6172)
/note="P1; derived from pRL1383a in INSD accession
AF403426, a derivative of RSF1010 INSD accession M28829;
the P1 promoter is controlled by MobC and MobA provide
transcription of mobA/repB, mobB, and, probably,
E-F-repA-repC-operon"
/regulatory_class="promoter"
CDS 6163..6447
/codon_start=1
/product="mobC"
/label=mobC
/note="mobilization gene; derived from pRL1383a in INSD
accession AF403426, a derivative of RSF1010 INSD accession
M28829"
/protein_id="AII99703.1"
/translation="MVKGSNKAADRLAKLEEQRARINAEIQRERAREQQQERKNETRRK
VLVGAMILAKVNSSEWPEDRLMAAMDAYLERDHDRALFGLPPRQKDEPG"
rep_origin 6474..6868
/label=RSF1010 oriV
/note="replication origin of the broad-host-range plasmid
RSF1010; requires the RSF1010 RepA/B/C proteins for
replication (Scholz et al., 1989)"
terminator complement(7035..7078)
/label=bacterial terminator
/note="putative bacterial transcription terminator"
regulatory complement(7044..7072)
/note="TrrnC; rrnC terminator from Lorist6 INSD accession
X98450; Lorist6"
/regulatory_class="terminator"
misc_feature 7111..7116
/label=NheI
/note="NheI"
misc_feature 7117..7137
/note="GC; GC-adaptor for seamless assembly, flanking BHR
replicons or neutral sites; synthetic"
misc_feature 7135..7140
/label=ZraI
/note="ZraI"
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