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pMQ235 vector (V004512)

Basic Vector Information

Vector Name:
pMQ235
Length:
9568 bp
Type:
Shuttle vector
Replication origin:
R6K γ ori
Host:
Yeast
Source/Author:
Kalivoda EJ, Horzempa J, Stella NA, Sadaf A, Kowalski RP, Nau GJ, Shanks RM.
Promoter:
LEU2

pMQ235 vector Vector Map

pMQ2359568 bp400800120016002000240028003200360040004400480052005600600064006800720076008000840088009200oriTrrnB T2 terminatorrrnB T1 terminatorM13 fwdMCSM13 revlac operatorlac promoterCAP binding siteR6K gamma oricatenterokinase siteSCEIhygLEU2LEU2 promoterCEN/ARS

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 ℃.

pMQ235 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_32045        9568 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMQ235, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9568)
  AUTHORS   Kalivoda EJ, Horzempa J, Stella NA, Sadaf A, Kowalski RP, Nau GJ, 
            Shanks RM.
  TITLE     New vector tools with a hygromycin resistance marker for use with 
            opportunistic pathogens
  JOURNAL   Mol. Biotechnol. 48 (1), 7-14 (2011)
  PUBMED    20972648
REFERENCE   2  (bases 1 to 9568)
  AUTHORS   Kalivoda EJ, Horzempa JE, Sadaf AA, Stella NA, Nau GJ, Shanks RM.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-DEC-2009) Ophthalmology, University of Pittsburgh, EEI
            1015, 203 Lothrop St, Pittsburgh, PA 15213, USA
REFERENCE   3  (bases 1 to 9568)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9568)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. 
            Biotechnol."; date: "2011"; volume: "48"; issue: "1"; pages: "7-14"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-DEC-2009) Ophthalmology, University of Pittsburgh, EEI 1015, 203
            Lothrop St, Pittsburgh, PA 15213, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9568
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            complement(399..508)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     terminator      complement(929..956)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(1048..1134)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     1484..1500
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(1504..1560)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1570..1586)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1594..1610)
                     /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)."
     promoter        complement(1618..1648)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1663..1684)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      1969..2357
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(2886..3533)
                     /gene="cat"
                     /label=cat
                     /note="Chloramphenicol acetyltransferase from
                     Staphylococcus aureus. Accession#: P00485"
     CDS             5027..5041
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
     CDS             complement(5282..5974)
                     /gene="SCEI"
                     /label=SCEI
                     /note="Intron-encoded endonuclease I-SceI from
                     Saccharomyces cerevisiae (strain ATCC 204508 / S288c). 
                     Accession#: P03882"
     CDS             complement(6002..6997)
                     /gene="hyg"
                     /label=hyg
                     /note="Hygromycin-B 7''-O-kinase from Streptomyces 
                     hygroscopicus. Accession#: P09979"
     CDS             complement(7286..8377)
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
     promoter        complement(8390..8794)
                     /label=LEU2 promoter
     misc_feature    9065..9568
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"

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