Basic Vector Information
Basic Vector Information | |||
---|---|---|---|
Vector Name | pCVD001 | Antibiotic Resistance | Gentamycin |
Length | 3313 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. |
pCVD001 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 ℃.
pCVD001 vector Sequence
LOCUS Exported 3313 bp ds-DNA circular SYN 18-DEC-2018 DEFINITION Broad host range vector vector pCVD001, complete sequence. ACCESSION KM017942 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3313) 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 3313) 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 3313) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..3313 /organism="Broad host range vector vector pCVD001" /mol_type="other DNA" /note="aacC1 is a device that confers gentamicin (Gm) resistance" /db_xref="taxon:1531859" 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..1338) /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..734 /regulatory_class="promoter" /note="PaacC1; upstream region of aacC1 ORF including promoter and RBS; derived from pDAH346" promoter 518..546 /gene="intI1 (promoter lies within the coding sequence)" /label=Pc promoter /note="class 1 integron promoter" CDS 735..1268 /codon_start=1 /product="aacC1" /label=aacC1 /note="confers gentamycin resistance; gentamicin acetyltransferase; derived from pDAH346" /protein_id="AII99790.1" /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR EEVMHFDIDPSTAT" regulatory 1276..1302 /regulatory_class="terminator" /note="trpA T; trpA terminator from Lorist6 INSD accession X98450, one nt shorter than reference; Lorist6" misc_feature 1312..1317 /label=NheI /note="NheI" misc_feature complement(1318..1338) /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 1336..1341 /label=EcoRV /note="EcoRV" rep_origin complement(1507..2180) /direction=LEFT /note="colE1_pMA-RQ; origin of replication of colE1 found inderived from pMA-RQ GeneART vector" rep_origin complement(1566..2154) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(2325..3185) /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="AII99791.1" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(3186..3290) /gene="bla" /label=AmpR promoter regulatory complement(3186..3255) /regulatory_class="promoter" /note="Pbla; upstream region of the bla ORF including promoter and RBS; derived from pUC19"
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