Basic Vector Information
- Vector Name:
- pNG168
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8960 bp
- Type:
- Shuttle vector
- Replication origin:
- ori
- Source/Author:
- DasSarma S.
- Promoter:
- T3
pNG168 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 ℃.
pNG168 vector Sequence
LOCUS 40924_33147 8960 bp DNA circular SYN 18-DEC-2018
DEFINITION Shuttle vector pNG168, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8960)
AUTHORS DasSarma S.
TITLE Natural plasmids and plasmid vectors of halophiles
JOURNAL (in) DasSarma,S., Fleischmann,E.M., Robb,F.T., Place,A.R.,
Sowers,K.R. and Schreier,H.J. (Eds.); ARCHAEA, A LABORATORY MANUAL -
HALOPHILES: 241-250; Cold Spring Harbor Laboratory Press, Cold
Spring Harbor, NY (1995)
REFERENCE 2 (bases 1 to 8960)
AUTHORS DasSarma S.
TITLE Direct Submission
JOURNAL Submitted (06-MAY-2003) Center of Marine Biotechnology, University
of Maryland Biotechnology Institute, 701 E. Pratt Street, Suite 236,
Baltimore, MD 21202
REFERENCE 3 (bases 1 to 8960)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 8960)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "(in)
DasSarma,S., Fleischmann,E.M., Robb,F.T., Place,A.R., Sowers,K.R.
and Schreier,H.J. (Eds.)"; volume: " ARCHAEA, A LABORATORY MANUAL -
HALOPHILES"; pages: " 241-250; Cold Spring Harbor Laboratory Press,
Cold Spring Harbor, NY (1995"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(06-MAY-2003) Center of Marine Biotechnology, University of Maryland
Biotechnology Institute, 701 E. Pratt Street, Suite 236, Baltimore,
MD 21202"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8960
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 849..3878
/codon_start=1
/gene="repH"
/product="RepH"
/label=repH
/note="replication protein; necessary for replication in
haloarchaea"
/protein_id="AAP44985.1"
/translation="MHTPNQQQGIRKIVPGGTLSTAGITITEVTPRVTEWIPDLLEELL
PRSIQSVRKFIRQEDPEVLTHARYNTVYRRLQEETLRFDHQEWCSTTDIWSDAEAEAVE
YVESLVEFAVKYSDVDEDDLDELSEYHQQRCKSLKQTLTTISTGRGPLNAGLEALAKGP
VRLHDELDDAPQPITLVLDGELWSKLDDRGTGIRALAAIAVLGSTFDVRLVISPALDAA
IERRYPDWYDSHLRLTETRETSSVESAGGDGQPSAEQLEEAWEAIQNLPEESGRLRLLR
NLPIEGSRDYRDLKQDDEIDVQAGTVGRYILDLEELGLVDIDRRGQYNSASLTGLGQVA
VEQYVTTDYRVIHPTQSTLETHLTPTPQPQASTVYPARSDTREGDQPGTAEDWIAATGS
PSEGADYVQWLDGPSGVLDAWGMHQRYLAGRRDRGVTLVDDRIERFEDGRVSYLSCFDD
DLFVATQWGGPLPTLGRIAGALLSDKALSKILTPSRLGNQFEEINDAVVEQLDREAGEI
IRRGHQIGWFSEDEEDYDGWRERIGSVRSLCLQQVGELTNSDDVEARTELLRDLHGLVA
SATQLYYAAGVDVTINVRVPDTGMLISDERRLDDFLGFARYTIPKQSVYGIHSGYRMLL
EDRPEKLKRRLPYEVDDADSTMHLTASWVFSGSTMIDLHDDIEDAIEMETNEIREAIAN
GQESAPVMEIPVQIGNSYSAIRNHVEDYASAKNYQVAHQEDIHEGKQDLERLVRLFLRV
LGTEDRPHRACPHDVAEAMLHVAQSSRNYDFITVRDISYGLSNLPTKRLLPELPPTATK
LLKTLLDADDPMGRSEIIDTADISESSYDRYINELAAWDIIEPREIEGHRRWEAHLEPW
WTPQSDRDEPYADPDPDTGILYAEFPRDVASAVMCHLITHYDLPDLETAYLEGIQPGDD
IKALFDDHDRLRRWRPFLWGAFADSDKLERGPSGTAASDSTVVRLGQSPGPDTAQSSFQ
DVSETATQRDRLSQPSPGLD"
gene 849..3878
/gene="repH"
/label=repH
protein_bind 4431..4452
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4467..4497
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4505..4521
/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 4529..4545
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 4566..4584
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
misc_feature complement(4597..4704)
/label=MCS
/note="pBluescript multiple cloning site"
promoter complement(4713..4731)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4741..4757)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 4899..5354
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 5380..5484
/label=AmpR promoter
CDS 5485..6342
/label=AmpR
/note="beta-lactamase"
rep_origin 6516..7104
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7428..8636)
/gene="hmgA"
/label=hmgA
/note="3-hydroxy-3-methylglutaryl-coenzyme A reductase from
Haloferax volcanii (strain ATCC 29605 / DSM 3757 / JCM 8879
/ NBRC 14742 / NCIMB 2012 / VKM B-1768 / DS2). Accession#:
Q59468"
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