pMQ118 vector (V004526)

Basic Vector Information

Vector Name:
pMQ118
Antibiotic Resistance:
Kanamycin
Length:
5980 bp
Type:
Cloning vector
Replication origin:
R6K γ ori
Host:
Yeast
Source/Author:
Shanks RM, Kadouri DE, MacEachran DP, O'Toole GA.
Promoter:
URA3

pMQ118 vector Vector Map

pMQ1185980 bp60012001800240030003600420048005400oriTrrnB T2 terminatorrrnB T1 terminatorM13 fwdMCSM13 revlac operatorlac promoterCAP binding siteT7 promoterR6K gamma oriNeoR/KanRrpsLURA3URA3 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 ℃.

pMQ118 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_31975        5980 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMQ118, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5980)
  AUTHORS   Shanks RM, Kadouri DE, MacEachran DP, O'Toole GA.
  TITLE     New yeast recombineering tools for bacteria
  JOURNAL   Plasmid 62 (2), 88-97 (2009)
  PUBMED    19477196
REFERENCE   2  (bases 1 to 5980)
  AUTHORS   Shanks RM, MacEachran DP, Powers SZ, Kadouri DE, O'Toole GA.
  TITLE     In vivo recombination vectors for modification and expression of 
            genes in Gram-negative and Gram-positive bacteria
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 5980)
  AUTHORS   Shanks RM, MacEachran DP, Powers SZ, Kadouri DE, O'Toole GA.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-MAR-2008) Ophthalmology, University of Pittsburgh, 203
            Lothrop St., Pittsburgh, PA 15213, USA
REFERENCE   4  (bases 1 to 5980)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 5980)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid"; 
            date: "2009"; volume: "62"; issue: "2"; pages: "88-97"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (03-MAR-2008) Ophthalmology, University of Pittsburgh, 203 Lothrop 
            St., Pittsburgh, PA 15213, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            complement(352..461)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     terminator      complement(882..909)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(1001..1087)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     1437..1453
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(1457..1513)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1523..1539)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1547..1563)
                     /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(1571..1601)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1616..1637)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        complement(1688..1706)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      1914..2302
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             2653..3444
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             3744..4115
                     /gene="rpsL"
                     /label=rpsL
                     /note="Small ribosomal subunit protein uS12 from Salmonella
                     paratyphi A (strain ATCC 9150 / SARB42). Accession#: 
                     Q5PIW1"
     CDS             complement(4381..5181)
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(5182..5402)
                     /label=URA3 promoter
     misc_feature    5430..5933
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"

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