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pKEW303-AcrVA5 vector (V012806#)

Price Information

Cat No. Plasmid Name Availability Add to cart
V012806 pKEW303-AcrVA5 In stock (lyophilized plasmid)

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Two vials of lyophilized plasmid will be delivered, each vial is about 5µg.

Basic Vector Information

Expresses AcrVA5 in bacteria, under TetR control.

Basic Vector Information
Vector Name pKEW303-AcrVA5 Antibiotic Resistance Ampicillin
Length 3198 bp Type E.coli expression plasmid
Source Jennifer Doudna Copy Number High copy number
Promoter tetR/tetA promoters

pKEW303-AcrVA5 vector Vector Map

pKEW303-AcrVA53198 bp6001200180024003000tetR/tetA promotersanti-CRISPR N-acetyltransferase AcrVA5rrnB T2 terminatororiAmpRAmpR promoterTetR

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 ℃.

pKEW303-AcrVA5 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                3198 bp ds-DNA     circular SYN 31-AUG-2022
DEFINITION  Expresses AcrVA5 in bacteria, under TetR control..
ACCESSION   .
VERSION     .
KEYWORDS    pKEW303-AcrVA5
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3198)
  AUTHORS   Watters KE, Fellmann C, Bai HB, Ren SM, Doudna JA
  TITLE     Systematic discovery of natural CRISPR-Cas12a inhibitors.
  JOURNAL   Science. 2018 Oct 12;362(6411):236-239. doi: 
            10.1126/science.aau5138. Epub 2018 Sep 6.
  PUBMED    30190307
REFERENCE   2  (bases 1 to 3198)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3198
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..56
                     /gene="tetR"
                     /note="tetR/tetA promoters"
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    37..55
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             83..361
                     /codon_start=1
                     /note="anti-CRISPR N-acetyltransferase AcrVA5"
                     /translation="MKIELSGGYICYSIEEDEVTIDMVEVTTKRQGIGSQLIDMVKDVA
                     REVGLPIGLYAYPQDDSISQEDLIEFYFSNDFEYDPDDVDGRLMRWS"
     terminator      594..621
                     /note="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      complement(747..1335)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1506..2366)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2367..2471)
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             complement(2557..3180)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /note="TetR"
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"