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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