pMD.SD vector (V004801)

MSDS

Basic Vector Information

Vector Name:: pMD.SD

Antibiotic Resistance:: Kanamycin

Length:: 3690 bp

Type:: Plasmid vector

Replication origin:: R6K γ ori

Source/Author:: Dharmasena MN, Stark CEC., Stibitz S, Kopecko DJ.

pMD.SD vector Vector Map

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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 vector Sequence

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LOCUS       40924_30411        3690 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Plasmid vector pMD.SD, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3690)
  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 3690)
  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 3690)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3690)
  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..3690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             411..1202
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     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="AJS13700.1"
                     /translation="MSITLSFQNASMNKLFEKECRNVATRALKYVRQKKTEGRLDEALS
                     VLISLKRIEPDVSRLMREYKQIIRLFNESRKDGGSTITSYEHLDYAKKLLVFDSENAYA
                     LKYAALNAMHLRDYTQALQYWQRLEKVNGPTEPVTRQISTCITALQKNTSGKS"
     protein_bind    complement(2023..2070)
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(2264..2632)
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(2665..2774)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      2934..3322
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     terminator      3458..3544
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3636..3663
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"

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