Basic Vector Information
- Vector Name:
- pL_delta_NG
- Antibiotic Resistance:
- Tetracycline
- Length:
- 6793 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Kobayashi H.
- Promoter:
- PLtetO-1
pL_delta_NG 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 ℃.
pL_delta_NG vector Sequence
LOCUS 40924_29576 6793 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pL_delta_NG DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6793)
AUTHORS Kobayashi H.
TITLE Inducible suppression of global translation by overuse of rare
codons
JOURNAL Appl. Environ. Microbiol. 81 (7), 2544-2553 (2015)
PUBMED 25636849
REFERENCE 2 (bases 1 to 6793)
AUTHORS Kobayashi H.
TITLE Direct Submission
JOURNAL Submitted (07-JAN-2015) Contact:Hideki Kobayashi Agency for
Marine-Earth Science and Technology, Institute of Biogeosciences;
2-15 Natsushima, Yokosuka, Kanagawa 237-0061, Japan URL
:http://www.jamstec.go.jp/j/
REFERENCE 3 (bases 1 to 6793)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6793)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
Environ. Microbiol."; date: "2015"; volume: "81"; issue: "7"; pages:
"2544-2553"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(07-JAN-2015) Contact:Hideki Kobayashi Agency for Marine-Earth
Science and Technology, Institute of Biogeosciences; 2-15
Natsushima, Yokosuka, Kanagawa 237-0061, Japan URL
:http://www.jamstec.go.jp/j/"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6793
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(22..1101)
/codon_start=1
/label=lacI
/note="lac repressor"
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
CDS complement(1317..1382)
/codon_start=1
/label=lambda N peptide
/note="N-terminal RNA-binding domain from the lambda
bacteriophage antiterminator protein N (Baron-Benhamou et
al., 2004)"
/translation="MDAQTRRRERRAEKQAQWKAAN"
promoter 1723..1752
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 1760..1776
/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)."
CDS 1796..2332
/codon_start=1
/gene="lgfp_delta_1"
/product="GFP deletion mutant"
/label=lgfp_delta_1
/protein_id="BAQ25584.1"
/translation="MSKGEELFTGVVPILVELDGDVNGQKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFYKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKMEYNYNSHNVYIMADKPKNGIKV
NFKIRHNIKDGSVQL"
gene 1796..2332
/gene="lgfp_delta_1"
/label=lgfp_delta_1
terminator 2543..2629
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2721..2748
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 2768..2859
/label=AmpR promoter
CDS 2860..3717
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
terminator 3750..3844
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
rep_origin 3932..4520
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4744..5457)
/codon_start=1
/label=yeGFP
/note="yeast-enhanced green fluorescent protein"
/translation="MSKGEELFTGVVPILVELDGDVNGQKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFYKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKMEYNYNSHNVYIMADKPKNGIKV
NFKIRHNIKDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMILLE
FVTAAGITHGMDELYK"
promoter complement(5482..5555)
/label=PLtetO-1 promoter
/note="modified phage lambda PL promoter with tet operator
sites (Lutz and Bujard, 1997)"
CDS 5859..5924
/codon_start=1
/label=lambda N peptide
/note="N-terminal RNA-binding domain from the lambda
bacteriophage antiterminator protein N (Baron-Benhamou et
al., 2004)"
/translation="MDAQTRRRERRAEKQAQWKAAN"
CDS 6140..6760
/codon_start=1
/label=TetR
/note="tetracycline repressor TetR"
/translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
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