Basic Vector Information
- Vector Name:
- pMDE7
- Length:
- 9703 bp
- Type:
- Mutator vector
- Replication origin:
- ori
- Source/Author:
- Sekurova ON, Sun YQ, Zehl M, Ruckert C
pMDE7 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 ℃.
pMDE7 vector Sequence
LOCUS 62056_16695 9703 bp DNA circular SYN 19-JUL-2021
DEFINITION Mutator vector pMDE7, complete sequence.
ACCESSION MW810320
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9703)
AUTHORS Sekurova ON, Sun YQ, Zehl M, Ruckert C, Stich A, Busche T,
Kalinowski J, Zotchev SB.
TITLE Coupling of the engineered DNA 'mutator' to a biosensor as a new
paradigm for activation of silent biosynthetic gene clusters in
Streptomyces
JOURNAL Nucleic Acids Res. (2021) In press
PUBMED 34197612
REFERENCE 2 (bases 1 to 9703)
AUTHORS Sekurova ON, Sun Y-Q., Zehl M, Ruckert C, Stich A, Busche T,
Kalinowski J, Zotchev SB.
TITLE Direct Submission
JOURNAL Submitted (09-MAR-2021) CeBiTec, Bielefeld University,
Universitaetsstr. 27, Bielefeld, NRW 33615, Germany
REFERENCE 3 (bases 1 to 9703)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res. (2021) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(09-MAR-2021) CeBiTec, Bielefeld University, Universitaetsstr. 27,
Bielefeld, NRW 33615, Germany"
COMMENT ##Assembly-Data-START##
Assembly Method :: canu v. 2.1.1
Sequencing Technology :: Illumina; ONT
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..9703
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory 1..40
/regulatory_class="promoter"
gene 99..3638
/gene="dnaE1"
/label=dnaE1
CDS 99..3638
/codon_start=1
/transl_table=11
/gene="dnaE1"
/product="DnaE1"
/label=dnaE1
/protein_id="QXN60066.1"
/translation="MSKPPFTHLHVHTQYSLLNGAARLKDMFDACNEMGMSHIAMSDHG
NLHGAYDFFHSAKKAGVTPIIGIEAYVAPESRRNKRKIQWGQPHQKRDDVSGSGGYTHK
TMWATNSKGLHNLFRLSSDAYAEGWLQKWPRMDKETISQWSEGIVASTGGPSGEVQTRL
RLGHFDEALKAAADYQDIFGKDRYFLELMDHGIEIEHRVRDGLLEIGRKLGIPPLVTND
SHYTYAHEATAHDALLCIQTGKNLSDPDRFRFDGTGYYLKSTDEMYAIDSSDAWQEGCA
NTRLIAEMIDTTGMFEKRDLMPKFDIPEGFTEITWFQEEVRRGMERRFPGGVPEDRQKQ
AEYEMDVIIQMGFPGYFLVVADFIMWAKNQGIAVGPGRGSAAGSIVAYAMGITDLDPIP
HGLIFERFLNPERVSMPDVDIDFDERRRVEVIRYVTEKYGADKVAMIGTYGKIKAKNAI
KDSARVLGYPYAMGDRLTKAMPADVLGKGIDLNGITDPTHPRYSEAGEIRSMYESEPDV
KKVIDTAKGVEGLVRQMGVHAAGVIMSSEPIVDHAPIWVRHTDGVTITQWDYPQCESLG
LLKMDFLGLRNLTIMDDAVKMVKSNKGIDLDLLSLPLDDPTTFELLQRGDTLGVFQFDG
GPMRSLLRLMKPDNFEDISAVSALYRPGPMGMDSHTNYALRKNGLQEITPIHKELEEPL
QEVLAVTYGLIVYQEQVQKAAQIIAGYSLGEADILRRVMGKKKPDELAKNFVLFQAGAR
KNGYSDEAIQALWDVLVPFAGYAFNKAHSAAYGLVSYWTGYLKANYPAEYMAALLTSVK
DDKDKSAVYLNECRRMGIKVLPPNVNESMSNFAAQGDDVILFGLSAVRNVGTNVVESII
KCRKAKGKYASFPDYLDKVEAVVCNKRTTESLIKAGAFDEMGHTRKGLTAQYEPMIDNV
VAVKRKEAEGQFDLFGGMGDEQSDEPGFGLDVVFGEDEWDKTYLLAQEREMLGLYVSDH
PLFGLEHVLSDKADAGISQLTGGDFGDGAVVTIGGIISGLQRKMTKQGNAWAIATVEDL
AGSLECMFFPATYQLVSTQLVEDAVVFVKGRLDKREDVPRLVAMELMIPDLSNAGTNAP
VVLTIPATRITPPMVSRLGEILTHHRGDSEVRIKLQGPTKTTVLRLDRHRVKPDPALFG
DLKVLLGPSCLAG"
gene complement(3758..3770)
/gene="lacZa'"
/label=lacZa'
/note="lacZ alpha peptide"
primer_bind complement(3771..3787)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3795..3811)
/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(3819..3849)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3864..3885)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4173..4761)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 4948..5943
/gene="hyg"
/label=hyg
/note="Hygromycin-B 7''-O-kinase from Streptomyces
hygroscopicus. Accession#: P09979"
oriT 6240..6349
/label=oriT
/note="incP origin of transfer"
CDS 6382..6750
/label=traJ
/note="oriT-recognizing protein"
rep_origin complement(7407..8810)
/direction=LEFT
/label=minimal SCP2* replicon
/note="minimal SCP2* replicon"
primer_bind 9521..9537
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
This page is informational only.