Basic Vector Information
- Vector Name:
- pMD.SD.Gad
- Antibiotic Resistance:
- Kanamycin
- Length:
- 9376 bp
- Type:
- Plasmid vector
- Replication origin:
- R6K γ ori
- Source/Author:
- Dharmasena MN, Stark CEC., Stibitz S, Kopecko DJ.
- Promoter:
- araBAD
pMD.SD.Gad 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 ℃.
pMD.SD.Gad vector Sequence
LOCUS 40924_30406 9376 bp DNA circular SYN 18-DEC-2018
DEFINITION Plasmid vector pMD.SD.Gad, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9376)
AUTHORS Dharmasena MN, Stark CEC., Stibitz S, Kopecko DJ.
TITLE Acid-resistant, attenuated microbial vector for improved oral
delivery of multiple targeted antigens
JOURNAL Patent: US WO/2015/021147-B 12-FEB-2015; FDA-Center for Biologics
Evaluation and Research; NIH campus Bldg. 29, 8800 Rockville Pike;
Laboratory of Enteric and Sexually Transmitted Diseases; Bethesda,
MD; USA;
REFERENCE 2 (bases 1 to 9376)
AUTHORS Dharmasena MN, Stark CEC., Stibitz S, Kopecko DJ.
TITLE Direct Submission
JOURNAL Submitted (16-MAY-2014) Laboratory of Enteric and Sexually
Transmitted Diseases, FDA-Center for Biologics Evaluation and
Research, NIH Campus Bldg. 29, 8800 Rockville Pike, Bethesda, MD
20892, USA
REFERENCE 3 (bases 1 to 9376)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9376)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Patent: US
WO/2015/021147-B 12-FEB-2015; FDA-Center for Biologics Evaluation
and Research; NIH campus Bldg. 29, 8800 Rockville Pike; Laboratory
of Enteric and Sexually Transmitted Diseases; Bethesda, MD; USA;"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(16-MAY-2014) Laboratory of Enteric and Sexually Transmitted
Diseases, FDA-Center for Biologics Evaluation and Research, NIH
Campus Bldg. 29, 8800 Rockville Pike, Bethesda, MD 20892, 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..9376
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 411..1202
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
protein_bind complement(1396..1429)
/label=FRT (minimal)
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
CDS 1500..1973
/codon_start=1
/product="tviD"
/label=tviD
/protein_id="AJS13705.1"
/translation="MSITLSFQNASMNKLFEKECRNVATRALKYVRQKKTEGRLDEALS
VLISLKRIEPDVSRLMREYKQIIRLFNESRKDGGSTITSYEHLDYAKKLLVFDSENAYA
LKYAALNAMHLRDYTQALQYWQRLEKVNGPTEPVTRQISTCITALQKNTSGKS"
CDS complement(2043..2918)
/label=araC
/note="L-arabinose regulatory protein"
promoter 2945..3229
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
CDS 3257..4654
/gene="gadA"
/label=gadA
/note="Glutamate decarboxylase alpha from Escherichia coli
(strain K12). Accession#: P69908"
CDS 4706..6103
/gene="gadB"
/label=gadB
/note="Glutamate decarboxylase beta from Escherichia coli
(strain K12). Accession#: P69910"
CDS 6128..7660
/gene="gadC"
/label=gadC
/note="Glutamate/gamma-aminobutyrate antiporter from
Escherichia coli (strain K12). Accession#: P63235"
protein_bind complement(7709..7756)
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS complement(7950..8318)
/label=traJ
/note="oriT-recognizing protein"
oriT complement(8351..8460)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
rep_origin 8620..9008
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
terminator 9144..9230
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 9322..9349
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
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