Basic Vector Information
- Vector Name:
- pWM91-L6-PT3-T4
- Antibiotic Resistance:
- Ampicillin
- Length:
- 13274 bp
- Type:
- Cloning vector
- Replication origin:
- R6K γ ori
- Source/Author:
- Lee JW, Gyorgy A, Cameron DE, Pyenson N, Choi KR, Way JC, Silver PA, Del Vecchio D, Collins JJ.
- Promoter:
- sacB
pWM91-L6-PT3-T4 vector Map
pWM91-L6-PT3-T4 vector Sequence
LOCUS 40924_46598 13274 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pWM91-L6-PT3-T4, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13274)
AUTHORS Lee JW, Gyorgy A, Cameron DE, Pyenson N, Choi KR, Way JC, Silver PA,
Del Vecchio D, Collins JJ.
TITLE Creating Single-Copy Genetic Circuits
JOURNAL Mol. Cell 63 (2), 329-336 (2016)
PUBMED 27425413
REFERENCE 2 (bases 1 to 13274)
AUTHORS Lee JW, Gyorgy A, Cameron DE, Pyenson N, Choi KR, Way JC, Silver PA,
Del Vecchio D, Collins JJ.
TITLE Direct Submission
JOURNAL Submitted (18-MAY-2016) Biological Engineering, MIT, 45 Carleton St,
MIT Building E25-Rm302, Cambridge, MA 02142, USA
REFERENCE 3 (bases 1 to 13274)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 13274)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Cell";
date: "2016"; volume: "63"; issue: "2"; pages: "329-336"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(18-MAY-2016) Biological Engineering, MIT, 45 Carleton St, MIT
Building E25-Rm302, Cambridge, MA 02142, 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..13274
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(60..515)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 657..673
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 683..701
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 727..743
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 776..1194
/label=similar to lacZ (upstream)
/note="similar to lacZ (upstream)"
terminator complement(1237..1331)
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(1394..2107)
/label=yeGFP
/note="yeast-enhanced green fluorescent protein"
regulatory complement(2108..2131)
/label=G5
/note="G5"
/regulatory_class="ribosome_binding_site"
RBS 2113..2121
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
promoter complement(2149..2222)
/label=PLtetO-1 promoter
/note="modified phage lambda PL promoter with tet operator
sites (Lutz and Bujard, 1997)"
misc_feature complement(2265..2348)
/label=mfLon-specific tag pdt2
/note="mfLon-specific tag pdt2"
CDS complement(2349..3428)
/label=lacI
/note="lac repressor"
RBS 3434..3442
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
promoter complement(3474..3547)
/label=PLtetO-1 promoter
/note="modified phage lambda PL promoter with tet operator
sites (Lutz and Bujard, 1997)"
protein_bind 3566..3585
/label=lac operator (symmetric)
/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). The
symmetric lac operator was optimized for tight binding of
lac repressor."
protein_bind 3602..3621
/label=lac operator (symmetric)
/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). The
symmetric lac operator was optimized for tight binding of
lac repressor."
promoter 3622..3651
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind complement(3658..3677)
/label=lac operator (symmetric)
/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). The
symmetric lac operator was optimized for tight binding of
lac repressor."
regulatory 3690..3721
/label=T4
/note="T4"
/regulatory_class="ribosome_binding_site"
RBS 3708..3716
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
CDS 3722..4342
/label=TetR
/note="tetracycline repressor TetR"
regulatory 4388..4462
/label=Ptrc-2
/note="Ptrc-2"
/regulatory_class="promoter"
promoter 4401..4430
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 4438..4454
/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)."
prim_transcript 4438
/label=transcription start
/note="transcription start"
regulatory 4470..4496
/label=M2
/note="M2"
/regulatory_class="ribosome_binding_site"
RBS 4483..4491
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
CDS 4497..5204
/label=mCherry
/note="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
terminator 5267..5353
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
misc_feature 5422..5821
/label=similar to lacZ (downstream)
/note="similar to lacZ (downstream)"
promoter complement(5840..5858)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(5879..5895)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 5903..5919
/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)."
promoter complement(5927..5957)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5972..5993)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(6697..7065)
/label=traJ
/note="oriT-recognizing protein"
oriT complement(7098..7207)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
promoter 8374..8819
/label=sacB promoter
/note="sacB promoter and control region"
CDS 8820..10238
/label=SacB
/note="secreted levansucrase that renders bacterial growth
sensitive to sucrose"
CDS complement(11811..12668)
/label=AmpR
/note="beta-lactamase"
promoter complement(12669..12773)
/label=AmpR promoter
rep_origin complement(12883..13271)
/direction=LEFT
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
This page is informational only.