pToxin-ecoRI vector (V002567)

Basic Vector Information

Vector Name:
pToxin-ecoRI
Antibiotic Resistance:
Ampicillin
Length:
3182 bp
Type:
Cloning vector
Replication origin:
p15A ori
Source/Author:
Chan CT, Lee JW, Cameron DE, Bashor CJ, Collins JJ.

pToxin-ecoRI vector Vector Map

pToxin-ecoRI3182 bp6001200180024003000lambda t0 terminatorT0RBSecoRIRrrnB T1 terminatorp15A orilambda t0 terminatorAmpRAmpR 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 ℃.

pToxin-ecoRI vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_43803        3182 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pToxin-ecoRI, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3182)
  AUTHORS   Chan CT, Lee JW, Cameron DE, Bashor CJ, Collins JJ.
  TITLE     'Deadman' and 'Passcode' microbial kill switches for bacterial 
            containment
  JOURNAL   Nat. Chem. Biol. 12 (2), 82-86 (2016)
  PUBMED    26641934
REFERENCE   2  (bases 1 to 3182)
  AUTHORS   Chan CTY., Lee JW, Cameron DE, Bashor CJ, Collins JJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-OCT-2015) Department of Biological Engineering, 
            Massachusetts Institute of Technology, 77 Massachusetts Ave., 
            E25-302b, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 3182)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3182)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem. 
            Biol."; date: "2016"; volume: "12"; issue: "2"; pages: "82-86"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-OCT-2015) Department of Biological Engineering, Massachusetts 
            Institute of Technology, 77 Massachusetts Ave., E25-302b, Cambridge,
            MA 02139, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3182
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      14..108
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     regulatory      115..189
                     /label=T0
                     /note="T0"
                     /regulatory_class="promoter"
     regulatory      195..214
                     /label=RBS
                     /note="RBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             221..1051
                     /gene="ecoRIR"
                     /label=ecoRIR
                     /note="Type II restriction enzyme EcoRI from Escherichia
                     coli. Accession#: P00642"
     terminator      1079..1165
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     rep_origin      1286..1831
                     /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."
     terminator      complement(1993..2087)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2120..2977)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(2978..3082)
                     /label=AmpR promoter

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