pMGK vector (V004745)

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 Map

pMGK5363 bp6001200180024003000360042004800p15A oriargUlacIq promoterlacICAP binding sitelambda t0 terminatorKanR

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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