pBAC-L6-PT3-T4 vector (V009337)

Basic Vector Information

Vector Name:
pBAC-L6-PT3-T4
Antibiotic Resistance:
Tetracycline
Length:
10509 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-T4 vector Map

pBAC-L6-PT3-T410509 bp5001000150020002500300035004000450050005500600065007000750080008500900095001000010500transcription startsopBsopAincCrepEori2oriVcat promoterCmRlambda t0 terminatoryeGFPG5PLtetOmfLon-specific tag pdt2lacIRBSPLtetO-1 promoterlac operator (symmetric)lac operator (symmetric)trc promoterlac operator (symmetric)T4TetRPtrc-2M2mCherryrrnB T1 terminator

pBAC-L6-PT3-T4 vector Sequence

LOCUS       40924_5514       10509 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBAC-L6-PT3-T4, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10509)
  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 10509)
  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 10509)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10509)
  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..10509
                     /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"
     promoter        5440..5542
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             5543..6199
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     terminator      complement(6337..6431)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6494..7207)
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
     regulatory      complement(7208..7231)
                     /label=G5
                     /note="G5"
                     /regulatory_class="ribosome_binding_site"
     RBS             7213..7221
                     /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)"
     regulatory      complement(7249..7322)
                     /label=PLtetO
                     /note="PLtetO"
                     /regulatory_class="promoter"
     promoter        complement(7249..7322)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"
     prim_transcript 7268
                     /label=transcription start
                     /note="transcription start"
     protein_bind    7304..7322
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    complement(7365..7448)
                     /label=mfLon-specific tag pdt2
                     /note="mfLon-specific tag pdt2"
     CDS             complement(7449..8528)
                     /label=lacI
                     /note="lac repressor"
     RBS             8534..8542
                     /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(8574..8647)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"
     protein_bind    8666..8685
                     /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    8702..8721
                     /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        8722..8751
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    complement(8758..8777)
                     /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      8790..8821
                     /label=T4
                     /note="T4"
                     /regulatory_class="ribosome_binding_site"
     RBS             8808..8816
                     /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             8822..9442
                     /label=TetR
                     /note="tetracycline repressor TetR"
     regulatory      9488..9562
                     /label=Ptrc-2
                     /note="Ptrc-2"
                     /regulatory_class="promoter"
     promoter        9501..9530
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    9538..9554
                     /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 9538
                     /label=transcription start
                     /note="transcription start"
     regulatory      9570..9596
                     /label=M2
                     /note="M2"
                     /regulatory_class="ribosome_binding_site"
     RBS             9583..9591
                     /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             9597..10304
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     terminator      10367..10453
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"

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