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pOsym(core)_mKate2 vector (V016464)

Basic Vector Information

Vector Name:
pOsym(core)_mKate2
Antibiotic Resistance:
Kanamycin
Length:
4217 bp
Type:
Cloning vector
Replication origin:
pSC101 ori
Source/Author:
Groseclose TM, Hersey AN, Huang BD, Wilson CJ.

pOsym(core)_mKate2 vector Vector Map

pOsym(core)_mKate24217 bp600120018002400300036004200lac operator (symmetric)trc promoter -10 hexamerstrong bacterial RBSmKate2rrnB T1 terminatorRep101pSC101 orilambda t0 terminatorNeoR/KanR

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 ℃.

pOsym(core)_mKate2 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       62056_18305        4217 bp DNA     circular SYN 16-AUG-2021
DEFINITION  Cloning vector pOsym(core)_mKate2, complete sequence.
ACCESSION   MZ198786
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4217)
  AUTHORS   Groseclose TM, Hersey AN, Huang BD, Wilson CJ.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   2  (bases 1 to 4217)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (13-MAY-2021) Chemical "
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    55..74
                     /label=lac operator (symmetric)
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     regulatory      74..79
                     /label=trc promoter -10 hexamer
                     /note="trc promoter -10 hexamer"
                     /regulatory_class="promoter"
     regulatory      98..109
                     /label=strong bacterial RBS
                     /note="strong bacterial RBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             116..811
                     /label=mKate2
                     /note="monomeric far-red fluorescent protein (Shcherbo et
                     al., 2009)"
     terminator      826..912
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(1453..2400)
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
     rep_origin      complement(2448..2670)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(3162..3256)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3290..4081)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"

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