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pET-28a-EGFP-N vector (V007512)

Basic Vector Information

Vector Name:
pET-28a-EGFP-N
Antibiotic Resistance:
Kanamycin
Length:
5990 bp
Type:
E. coli Expression Vectors, Fluorescent Protein Re
Replication origin:
ori
Selection Marker:
N-EGFP
Promoter:
T7/lac

pET-28a-EGFP-N vector Vector Map

pET-28a-EGFP-N5990 bp60012001800240030003600420048005400f1 oriKanRoribomropCAP binding sitelacIlacI promoterT7 promoterlac operatorRBSEGFP6xHisT7 terminator

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 ℃.

pET-28a-EGFP-N vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_17816        5990 bp DNA     circular SYN 13-JAN-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5990)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5990)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(563..1375)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      1497..2085
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2271..2413)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2518..2706)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(3481..3502)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3518..4597)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(4598..4675)
                     /label=lacI promoter
     promoter        4984..5002
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5003..5027
                     /label=lac operator
                     /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)."
     RBS             5042..5064
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5071..5787
                     /label=EGFP
                     /note="enhanced GFP"
     CDS             5834..5851
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      5918..5965
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"

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