Basic Vector Information
- Vector Name:
- pBAC-L6-PT3-T4Ei
- Antibiotic Resistance:
- Tetracycline
- Length:
- 10980 bp
- Type:
- Cloning vector
- Replication origin:
- ori2
- Source/Author:
- Lee JW, Gyorgy A, Cameron DE, Pyenson N, Choi KR, Way JC, Silver PA, Del Vecchio D, Collins JJ.
pBAC-L6-PT3-T4Ei vector Vector Map
pBAC-L6-PT3-T4Ei vector Sequence
LOCUS V009335 10980 bp DNA circular SYN 17-DEC-2018
DEFINITION Exported.
ACCESSION V009335
VERSION V009335
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 10980)
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 10980)
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 10980)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10980)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Cell";
date: "2016"; volume: "63"; issue: "2"; pages: "329-336"
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"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10980
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(105..1073)
/label="sopB"
/note="partitioning protein for the bacterial F plasmid"
CDS complement(1076..2248)
/label="sopA"
/note="partitioning protein for the bacterial F plasmid"
misc_feature complement(2574..2824)
/label="incC"
/note="incompatibility region of the bacterial F plasmid"
CDS complement(2830..3582)
/label="repE"
/note="replication initiation protein for the bacterial F
plasmid"
rep_origin complement(3673..3892)
/direction=LEFT
/label="ori2"
/note="secondary origin of replication for the bacterial F
plasmid; also known as oriS"
rep_origin complement(3968..4582)
/direction=LEFT
/label="oriV"
/note="origin of replication for the bacterial F plasmid"
misc_feature 5425..5472
/label="FRT site"
/note="FRT site"
protein_bind complement(5425..5472)
/label="FRT"
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
promoter 5496..5598
/label="cat promoter"
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 5599..6255
/label="CmR"
/note="chloramphenicol acetyltransferase"
protein_bind complement(6384..6417)
/label="FRT (minimal)"
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
protein_bind complement(6437..6663)
/label="HK022 attP"
/note="attachment site of phage HK022"
terminator complement(6717..6811)
/label="lambda t0 terminator"
/note="transcription terminator from phage lambda"
CDS complement(6862..7692)
/gene="ecoRIR"
/label="Type II restriction enzyme EcoRI"
/note="Type II restriction enzyme EcoRI from Escherichia
coli. Accession#: P00642"
regulatory complement(7720..7793)
/label="PLtetO"
/note="PLtetO"
/regulatory_class="promoter"
promoter complement(7720..7793)
/label="PLtetO-1 promoter"
/note="modified phage lambda PL promoter with tet operator
sites (Lutz and Bujard, 1997)"
prim_transcript 7739
/label="transcription start"
/note="transcription start"
protein_bind 7775..7793
/gene="tetO"
/label="tet operator"
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
misc_feature 7836..7919
/label="mfLon-specific tag pdt2"
/note="mfLon-specific tag pdt2"
CDS complement(7920..8999)
/label="lacI"
/note="lac repressor"
RBS 9005..9013
/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(9045..9118)
/label="PLtetO-1 promoter"
/note="modified phage lambda PL promoter with tet operator
sites (Lutz and Bujard, 1997)"
protein_bind 9137..9156
/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 9173..9192
/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 9193..9222
/label="trc promoter"
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind complement(9229..9248)
/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 9261..9292
/label="T4"
/note="T4"
/regulatory_class="ribosome_binding_site"
RBS 9279..9287
/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 9293..9913
/label="TetR"
/note="tetracycline repressor TetR"
promoter 9972..10001
/label="trc promoter"
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 10009..10025
/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)."
regulatory 10041..10067
/label="M2"
/note="M2"
/regulatory_class="ribosome_binding_site"
RBS 10054..10062
/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 10068..10775
/label="mCherry"
/note="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
terminator 10838..10924
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
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