Basic Vector Information
- Vector Name:
- pTXB1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6706 bp
- Type:
- Cloning Vectors
- Replication origin:
- ori
- Source/Author:
- Southworth MW, Amaya K, Evans TC, Xu MQ, Perler FB.
- Copy Number:
- High copy number
pTXB1 vector Vector Map
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 ℃.
pTXB1 vector Sequence
LOCUS pTXB1. 6706 bp DNA circular SYN 01-JAN-1980
DEFINITION Bacterial expression vector that encodes a thiol-cleavable
C-terminal intein followed by a chitin-binding domain.
ACCESSION .
VERSION .
KEYWORDS pTXB1.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6706)
AUTHORS Southworth MW, Amaya K, Evans TC, Xu MQ, Perler FB.
TITLE Purification of proteins fused to either the amino or carboxy
terminus of the Mycobacterium xenopi gyrase A intein.
JOURNAL BioTechniques 1999;27:110-4, 116, 118-20.
PUBMED 10407673
REFERENCE 2 (bases 1 to 6706)
AUTHORS New England Biolabs
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6706)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"BioTechniques"; date: "1999"; volume: "27"; pages: "110-4, 116,
118-20"
COMMENT SGRef: number: 2; type: "Journal Article"
COMMENT Clone between the NdeI and SapI sites to generate a fusion protein
that can be isolated with no added amino acids.
FEATURES Location/Qualifiers
source 1..6706
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 35..139
/label=AmpR promoter
CDS 140..997
/label=AmpR
/note="beta-lactamase"
rep_origin complement(1042..1555)
/direction=LEFT
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
rep_origin 1666..2254
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2626..2814)
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
protein_bind complement(3337..3358)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(3374..4453)
/label=lacI
/note="lac repressor"
promoter complement(4454..4531)
/label=lacI promoter
terminator 4684..4770
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 4867..4953
/gene="Escherichia coli rrnB"
/label=Escherichia coli rrnB terminator
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 5050..5136
/gene="Escherichia coli rrnB"
/label=Escherichia coli rrnB terminator
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 5233..5319
/gene="Escherichia coli rrnB"
/label=Escherichia coli rrnB terminator
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator complement(5419..5462)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
promoter 5637..5655
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 5656..5680
/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 5695..5717
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
misc_feature 5722..5775
/label=MCS
/note="MCS"
/note="multiple cloning site"
CDS 5725..5727
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
CDS 5776..6369
/label=Mxe GyrA intein
/note="modified GyrA intein (Southworth et al., 1999)"
CDS 6400..6555
/label=CBD
/note="chitin binding domain from chitinase A1 (Watanabe et
al., 1994)"
terminator 6634..6681
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
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