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pCVD001 vector (V001580#)

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

pCVD0013313 bp6001200180024003000M13 fwdT7 promoterEcoRVEcoRVcolE1_pMA-RQbla_pMA-RQAmpR promoter

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

Copy Sequence

Download GeneBank File(.gb)

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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