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pLysS vector (V010927)

Basic Vector Information

Vector Name:
pLysS
Antibiotic Resistance:
Chloramphenicol
Length:
4886 bp
Type:
pET & Duet Vectors (Novagen)
Replication origin:
p15A ori
Source/Author:
Novagen (EMD Millipore)
Copy Number:
Medium copy number

pLysS vector Vector Map

pLysS4886 bp6001200180024003000360042004800cat promoterp15A oritet promoterT7 Phi-3.8 promoterT7 lysozymeCmR

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 ℃.

pLysS vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_29536        4886 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Plasmid for expressing low levels of T7 lysozyme, an inhibitor of T7
            RNA polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4886)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 4886)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     The T7 lysozyme gene is not highly expressed because it is in the 
            antisense orientation relative to the tet promoter.
FEATURES             Location/Qualifiers
     source          1..4886
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(220..322)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      complement(848..1392)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     promoter        1504..1532
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     promoter        complement(1981..2014)
                     /label=T7 Phi-3.8 promoter
                     /note="weak promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2015..2467)
                     /codon_start=1
                     /label=T7 lysozyme
                     /note="lysozyme from bacteriophage T7"
                     /translation="MARVQFKQRESTDAIFVHCSATKPSQNVGVREIRQWHKEQGWLDV
                     GYHFIIKRDGTVEAGRDEMAVGSHAKGYNHNSIGVCLVGGIDDKGKFDANFTPAQMQSL
                     RSLLVTLLAKYEGAGLRAHHEVAPKACPSFDLKRWWEKNELVTSDRG"
     CDS             complement(join(4449..4886,1..219))
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"

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