pME3087 vector (V004781)

Basic Vector Information

Vector Name:
pME3087
Antibiotic Resistance:
Tetracycline
Length:
6822 bp
Type:
Suicide vector
Replication origin:
ori
Source/Author:
Scott TA, Heine D, Qin Z, Wilkinson B.

pME3087 vector Vector Map

pME30876822 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600TetRTcRExc2mbeAroporiToricolE1MCS

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 ℃.

pME3087 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_30530        6822 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Suicide vector pME3087, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6822)
  AUTHORS   Scott TA, Heine D, Qin Z, Wilkinson B.
  TITLE     An L-threonine transaldolase is required for 
            L-threo-beta-hydroxy-alpha-amino acid assembly during obafluorin 
            biosynthesis
  JOURNAL   Nat Commun 8, 15935 (2017)
  PUBMED    28649989
REFERENCE   2  (bases 1 to 6822)
  AUTHORS   Scott TA.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-SEP-2016) Molecular Microbiology, John Innes Centre, 
            Norwich Research Park, Norwich, Norfolk NR4 7UH, United Kingdom
REFERENCE   3  (bases 1 to 6822)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6822)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun 
            8, 15935 (2017)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-SEP-2016) Molecular Microbiology, John Innes Centre, Norwich 
            Research Park, Norwich, Norfolk NR4 7UH, United Kingdom"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6822
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(724..1371)
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
     CDS             1477..2673
                     /label=TcR
                     /note="tetracycline efflux protein"
     CDS             2679..2825
                     /codon_start=1
                     /gene="exc2"
                     /product="Exc2"
                     /label=exc2
                     /protein_id="AQZ26564.1"
                     /translation="METIGNSLRGTTQFAGTDYRSKDLTPKKSRLLADTISAVYLDGYE
                     GRQ"
     gene            2679..2825
                     /gene="exc2"
                     /label=exc2
     misc_feature    complement(2800..3083)
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to
                     maintain plasmids as monomers"
     CDS             complement(3039..4589)
                     /gene="mbeA"
                     /label=mbeA
                     /note="DNA relaxase MbeA from Escherichia coli. Accession#:
                     P13658"
     CDS             complement(4582..4926)
                     /gene="mbeC"
                     /label=mbeC
                     /note="Mobilization protein MbeC from Escherichia coli. 
                     Accession#: P13657"
     CDS             4966..5154
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     oriT            5242..5330
                     /note="oriT"
     rep_origin      complement(5574..6164)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             6369..6710
                     /codon_start=1
                     /gene="colE1"
                     /product="colicin E1 immunity protein"
                     /label=colE1
                     /protein_id="AQZ26569.1"
                     /translation="MSLRYYIKNILFGLYCTLIYIYLITKNSEGYYFLVSDKMLYAIVI
                     STILCPYSKYAIEYIAFNFIKKDFFERRKNLNNAPVAKLNLFMLYNLLCLVLAIPFGLL
                     GLFISIKNN"
     gene            6369..6710
                     /gene="colE1"
                     /label=colE1
     misc_feature    6766..6822
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"

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