Basic Vector Information
- Vector Name:
- pMGK
- Antibiotic Resistance:
- Kanamycin
- Length:
- 5363 bp
- Type:
- Cloning vector
- Replication origin:
- p15A ori
- Source/Author:
- Boel G, Letso R, Neely H, Price WN, Wong KH, Su M, Luff JD, Valecha M, Everett JK, Acton TB, Xiao R, Montelione GT, Aalberts DP, Hunt JF.
pMGK 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 ℃.
pMGK vector Sequence
LOCUS 40924_30790 5363 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pMGK, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5363)
AUTHORS Boel G, Letso R, Neely H, Price WN, Wong KH, Su M, Luff JD, Valecha
M, Everett JK, Acton TB, Xiao R, Montelione GT, Aalberts DP, Hunt
JF.
TITLE Codon influence on protein expression in E. coli correlates with
mRNA levels
JOURNAL Nature 529, 358-363 (2016)
PUBMED 26760206
REFERENCE 2 (bases 1 to 5363)
AUTHORS Boel G, Hunt JF, Montelione GT, Acton T.
TITLE Direct Submission
JOURNAL Submitted (23-JUN-2015) Biological Sciences, Columbia University,
1212 Amsterdam Ave., New York, NY 10027, USA
REFERENCE 3 (bases 1 to 5363)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5363)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nature 529,
358-363 (2016)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(23-JUN-2015) Biological Sciences, Columbia University, 1212
Amsterdam Ave., New York, NY 10027, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..5363
/mol_type="other DNA"
/organism="synthetic DNA construct"
tRNA complement(893..969)
/label=argU
promoter 1694..1771
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
CDS 1772..2851
/codon_start=1
/label=lacI
/note="lac repressor"
/translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
protein_bind 2867..2888
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
terminator 3056..3090
/label=lambda t0 terminator
/note="minimal transcription terminator from phage lambda
(Scholtissek and Grosse, 1987)"
CDS complement(3802..4614)
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin complement(join(5209..5363,1..391))
/direction=LEFT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin."
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