Basic Vector Information
- Vector Name:
- pLZ1
- Antibiotic Resistance:
- Kanamycin
- Length:
- 4712 bp
- Type:
- Cloning vector
- Replication origin:
- RSF ori
- Source/Author:
- Zhu L, Klenner C, Kuhn A, Dalbey RE.
pLZ1 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 ℃.
pLZ1 vector Sequence
LOCUS 40924_29541 4712 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pLZ1, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4712)
AUTHORS Zhu L, Klenner C, Kuhn A, Dalbey RE.
TITLE Both YidC and SecYEG are required for translocation of the
periplasmic loops 1 and 2 of the multispanning membrane protein TatC
JOURNAL J. Mol. Biol. 424 (5), 354-367 (2012)
PUBMED 23058713
REFERENCE 2 (bases 1 to 4712)
AUTHORS Zhu L, Wasey A, White SH, Dalbey RE.
TITLE Charge composition features of model single-span membrane proteins
that determine selection of YidC and SecYEG translocase pathways in
Escherichia coli
JOURNAL J. Biol. Chem. 288 (11), 7704-7716 (2013)
PUBMED 23355473
REFERENCE 3 (bases 1 to 4712)
AUTHORS Zhu L, Kaback HR, Dalbey RE.
TITLE YidC protein, a molecular chaperone for LacY protein folding via the
SecYEG protein machinery
JOURNAL J. Biol. Chem. 288 (39), 28180-28194 (2013)
PUBMED 23928306
REFERENCE 4 (bases 1 to 4712)
AUTHORS Soman R, Yuan J, Kuhn A, Dalbey RE.
TITLE The polarity and charge of the periplasmic loop determines the YidC
and Sec translocase requirement for the M13 procoat lep protein
JOURNAL J. Biol. Chem. (2013) In press
PUBMED 24275657
REFERENCE 5 (bases 1 to 4712)
AUTHORS Zhu L, Dalbey R.
TITLE Direct Submission
JOURNAL Submitted (29-NOV-2013) Department of Chemistry and Biochemistry,
The Ohio State University, RM580, W. 12th Ave., Columbus, OH 43210,
USA
REFERENCE 6 (bases 1 to 4712)
TITLE Direct Submission
REFERENCE 7 (bases 1 to 4712)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J. Mol.
Biol."; date: "2012"; volume: "424"; issue: "5"; pages: "354-367"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "J. Biol.
Chem."; date: "2013"; volume: "288"; issue: "11"; pages: "7704-7716"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "J. Biol.
Chem."; date: "2013"; volume: "288"; issue: "39"; pages:
"28180-28194"
COMMENT SGRef: number: 4; type: "Journal Article"; journalName: "J. Biol.
Chem. (2013) In press"
COMMENT SGRef: number: 5; type: "Journal Article"; journalName: "Submitted
(29-NOV-2013) Department of Chemistry and Biochemistry, The Ohio
State University, RM580, W. 12th Ave., Columbus, OH 43210, USA"
COMMENT SGRef: number: 6; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4712
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 20..38
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 39..63
/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)."
promoter 197..227
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
protein_bind 235..251
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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 complement(397..413)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
terminator 604..651
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
terminator 704..790
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 882..909
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 1009..1057
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
terminator 1119..1167
/label=fd terminator
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
CDS 1313..1357
/codon_start=1
/label=S-Tag
/note="affinity and epitope tag derived from pancreatic
ribonuclease A"
/translation="KETAAAKFERQHMDS"
terminator 1409..1456
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(1689..2501)
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
promoter complement(2502..2593)
/label=AmpR promoter
rep_origin complement(2609..3358)
/direction=LEFT
/label=RSF ori
/note="Plasmids containing the RSF 1030 origin of
replication can be propagated in E. coli cells that contain
additional plasmids with compatible origins."
protein_bind complement(3516..3537)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(3553..4632)
/codon_start=1
/label=lacI
/note="lac repressor"
/translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
promoter complement(4633..4710)
/label=lacI promoter
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