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pNG4 vector (V004330)

Basic Vector Information

Vector Name:
pNG4
Antibiotic Resistance:
Ampicillin
Length:
9791 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.

pNG4 vector Vector Map

pNG49791 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400880092009600AmpRAmpR promoteroriCAP binding sitelac promoterlac operatorM13 revfd terminatororiTtraJhygSCO4849

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 ℃.

pNG4 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_33167        9791 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pNG4, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9791)
  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 9791)
  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 9791)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9791)
  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..9791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(120..977)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(978..1041)
                     /label=AmpR promoter
     rep_origin      1310..1898
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2186..2207
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2222..2252
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2260..2276
                     /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     2284..2300
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      2463..2511
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     regulatory      complement(2822..2825)
                     /regulatory_class="ribosome_binding_site"
     oriT            5097..5206
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             5239..5607
                     /label=traJ
                     /note="oriT-recognizing protein"
     CDS             complement(6323..7318)
                     /gene="hyg"
                     /label=hyg
                     /note="Hygromycin-B 7''-O-kinase from Streptomyces 
                     hygroscopicus. Accession#: P09979"
     CDS             complement(8003..9313)
                     /codon_start=1
                     /gene="SCO4849"
                     /product="SCO4849 protein"
                     /label=SCO4849
                     /protein_id="AML61769.1"
                     /translation="MVVVFVLVALLMVGVLVTANWYVWRRLFRDTTRAPGPVRRIGAAV
                     IAGGWLLAVGALVAERAGAPFWLQRVLAWPGFLWLALSIYLLLAVLAGEVVRPLLRRFL
                     ERRAAARRTAGAEPSVTPAADTSRIPAGTAPPKTPEAPETPEAPGTEALAAQGSPLAVP
                     SRRLFVSRVVAGAAAAAAVGTVGYGTYGVLSGPKVKRVTVPLAKLPRAAHGFRIAVVSD
                     IHLGPVLGRGFAQQVVDTINSTQPDLIAVVGDLVDGSVKDLGPAAAPLARLTARHGAYF
                     VTGNHEYFSGAEQWVAEVRRLGLLPLENARTELPHFDLAGVNDVAGEDEGQGPDYDRAL
                     GDRDRSRACVLLAHQPVQIHDAVDHGVDLQLSGHTHGGQLWPGNLIAGAANPTLAGLER
                     YGDTQLYVSRGAGAWGPPTRVGAPSDITVIELASRQA"
     gene            complement(8003..9313)
                     /gene="SCO4849"
                     /label=SCO4849
     regulatory      complement(9368..9790)
                     /regulatory_class="promoter"

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