Basic Vector Information
| Basic Vector Information | |||
|---|---|---|---|
| Vector Name | pCVD004 | Antibiotic Resistance | Ampicillin |
| Length | 3659 bp | Type | Broad host range vector |
| Source | Taton A, Unglaub F, Wright NE, Zeng WY, Paz-Yepez J, Brahamsha B, Palenik B, Peterson TC, Haerizadeh F, Golden SS, Golden JW. | ||
pCVD004 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 ℃.
pCVD004 vector Sequence
LOCUS Exported 3659 bp ds-DNA circular SYN 18-DEC-2018
DEFINITION Broad host range vector vector pCVD004, complete sequence.
ACCESSION KM017939
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3659)
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 3659)
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 3659)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..3659
/organism="Broad host range vector vector pCVD004"
/mol_type="other DNA"
/note="bla_A7120 is a device that confers ampicillin (Ap)
resistance"
/db_xref="taxon:1531862"
primer_bind 293..309
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 320..338
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 370..375
/label=EcoRV
/note="EcoRV"
misc_feature complement(373..1684)
/note="CVD-ANTR; device for the GC-adaptor assembly:
antibiotic resistance markers"
misc_feature complement(373..393)
/note="C2G; GC-adaptor for seamless assembly, flanking
antibiotic resistance markers, functional devices or custom
sequences; synthetic"
misc_feature 394..399
/label=AgeI
/note="AgeI"
regulatory 400..539
/regulatory_class="promoter"
/note="PaadA; aadA 5'UTR, including promoter and RBS
conserved between pAM2178 and pcr8GWtopo"
CDS 540..1613
/codon_start=1
/product="bla_A7120"
/label=bla_A7120
/note="confers ampicillin resistance; codon optimized for
Anabaena sp. PCC7120; amino acid sequence derived from
pUC19"
/protein_id="AII99784.1"
/translation="MRSRNWSRTLTERSGGNGAVAVFMACYDCFFGVQSMPSKQQARYA
VGRCLMLWSSNDVTQQGSRPKTKLNIMSIQHFRVALIPFFAAFCLPVFAHPETLVKVKD
AEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRR
IHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLH
NMGDHVTRLDRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWME
ADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMD
ERNRQIAEIGASLIKHW"
CDS 753..1613
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
regulatory 1622..1648
/regulatory_class="terminator"
/note="trpA T; trpA terminator from Lorist6 INSD accession
X98450, one nt shorter than reference; Lorist6"
misc_feature 1658..1663
/label=NheI
/note="NheI"
misc_feature complement(1664..1684)
/note="GC; GC-adaptor for seamless assembly, flanking
antibiotic resistance markers or Escherichia coli origins
to be assembled with a cyanobacterial replicon; synthetic"
misc_feature 1682..1687
/label=EcoRV
/note="EcoRV"
rep_origin complement(1853..2526)
/direction=LEFT
/note="colE1_pMA-RQ; origin of replication of colE1 found
inderived from pMA-RQ GeneART vector"
rep_origin complement(1912..2500)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2671..3531)
/codon_start=1
/product="bla_pMA-RQ"
/label=bla_pMA-RQ
/note="confers ampicillin resistance; beta-lactamase;
derived from pMA-RQ GeneART vector, has one point mutation
compared to pUC19"
/protein_id="AII99785.1"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3532..3636)
/gene="bla"
/label=AmpR promoter
regulatory complement(3532..3601)
/regulatory_class="promoter"
/note="Pbla; upstream region of the bla ORF including
promoter and RBS; derived from pUC19"
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