Basic Vector Information
- Vector Name:
- pNG2
- Length:
- 8756 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Gonzalez-Quinonez N, Lopez-Garcia MT, Yague P, Rioseras B, Pisciotta A, Alduina R, Manteca A.
pNG2 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 ℃.
pNG2 vector Sequence
LOCUS 40924_33157 8756 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pNG2, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8756)
AUTHORS Gonzalez-Quinonez N, Lopez-Garcia MT, Yague P, Rioseras B, Pisciotta
A, Alduina R, Manteca A.
TITLE New PhiBT1 site-specific integrative vectors with neutral phenotype
in Streptomyces
JOURNAL Appl. Microbiol. Biotechnol. 100 (6), 2797-2808 (2016)
PUBMED 26758297
REFERENCE 2 (bases 1 to 8756)
AUTHORS Gonzalez-Quinonez N, Lopez-Garcia MT, Yague P, Rioseras B, Manteca
A.
TITLE Direct Submission
JOURNAL Submitted (15-APR-2015) Biologia Funcional, Universidad de Oviedo,
c/Julian Claveria s/n Facultad de Medicina, Oviedo 33006, Spain
REFERENCE 3 (bases 1 to 8756)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 8756)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
Microbiol. Biotechnol."; date: "2016"; volume: "100"; issue: "6";
pages: "2797-2808"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(15-APR-2015) Biologia Funcional, Universidad de Oviedo, c/Julian
Claveria s/n Facultad de Medicina, Oviedo 33006, Spain"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..8756
/mol_type="other DNA"
/organism="synthetic DNA construct"
oriT 2150..2259
/label=oriT
/note="incP origin of transfer"
CDS 2292..2660
/label=traJ
/note="oriT-recognizing protein"
CDS complement(3376..4371)
/gene="hyg"
/label=hyg
/note="Hygromycin-B 7''-O-kinase from Streptomyces
hygroscopicus. Accession#: P09979"
CDS complement(5056..6366)
/codon_start=1
/gene="SCO4849"
/product="SCO4849 protein"
/label=SCO4849
/protein_id="AML61763.1"
/translation="MVVVFVLVALLMVGVLVTANWYVWRRLFRDTTRAPGPVRRIGAAV
IAGGWLLAVGALVAERAGAPFWLQRVLAWPGFLWLALSIYLLLAVLAGEVVRPLLRRFL
ERRAAARRTAGAEPSVTPAADTSRIPAGTAPPKTPEAPETPEAPGTEALAAQGSPLAVP
SRRLFVSRVVAGAAAAAAVGTVGYGTYGVLSGPKVKRVTVPLAKLPRAAHGFRIAVVSD
IHLGPVLGRGFAQQVVDTINSTQPDLIAVVGDLVDGSVKDLGPAAAPLARLTARHGAYF
VTGNHEYFSGAEQWVAEVRRLGLLPLENARTELPHFDLAGVNDVAGEDEGQGPDYDRAL
GDRDRSRACVLLAHQPVQIHDAVDHGVDLQLSGHTHGGQLWPGNLIAGAANPTLAGLER
YGDTQLYVSRGAGAWGPPTRVGAPSDITVIELASRQA"
gene complement(5056..6366)
/gene="SCO4849"
/label=SCO4849
regulatory complement(6421..6843)
/regulatory_class="promoter"
rep_origin 7119..7707
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 7995..8016
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 8031..8061
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8069..8085
/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)."
primer_bind 8093..8109
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
terminator 8272..8320
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
regulatory complement(8631..8634)
/regulatory_class="ribosome_binding_site"
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