Basic Vector Information
- Vector Name:
- pBAC-L6-PT3-T4Ci
- Antibiotic Resistance:
- Tetracycline
- Length:
- 10452 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-T4Ci vector Vector Map
pBAC-L6-PT3-T4Ci vector Sequence
LOCUS 40924_5519 10452 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pBAC-L6-PT3-T4Ci, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10452)
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 10452)
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 10452)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10452)
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..10452
/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..7164)
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
regulatory complement(7171..7185)
/label=RBS
/note="RBS"
/regulatory_class="ribosome_binding_site"
regulatory complement(7192..7265)
/label=PLtetO
/note="PLtetO"
/regulatory_class="promoter"
promoter complement(7192..7265)
/label=PLtetO-1 promoter
/note="modified phage lambda PL promoter with tet operator
sites (Lutz and Bujard, 1997)"
prim_transcript 7211
/label=transcription start
/note="transcription start"
protein_bind 7247..7265
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
misc_feature complement(7308..7391)
/label=mfLon-specific tag pdt2
/note="mfLon-specific tag pdt2"
CDS complement(7392..8471)
/label=lacI
/note="lac repressor"
RBS 8477..8485
/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(8517..8590)
/label=PLtetO-1 promoter
/note="modified phage lambda PL promoter with tet operator
sites (Lutz and Bujard, 1997)"
protein_bind 8609..8628
/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 8645..8664
/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 8665..8694
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind complement(8701..8720)
/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 8733..8764
/label=T4
/note="T4"
/regulatory_class="ribosome_binding_site"
RBS 8751..8759
/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 8765..9385
/label=TetR
/note="tetracycline repressor TetR"
regulatory 9431..9505
/label=Ptrc-2
/note="Ptrc-2"
/regulatory_class="promoter"
promoter 9444..9473
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 9481..9497
/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 9481
/label=transcription start
/note="transcription start"
regulatory 9513..9539
/label=M2
/note="M2"
/regulatory_class="ribosome_binding_site"
RBS 9526..9534
/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 9540..10247
/label=mCherry
/note="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
terminator 10310..10396
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
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