pBAC-SC vector (V009332)

Basic Vector Information

Vector Name:
pBAC-SC
Antibiotic Resistance:
Tetracycline
Length:
10512 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-SC vector Map

pBAC-SC10512 bp5001000150020002500300035004000450050005500600065007000750080008500900095001000010500transcription startsopBsopAincCrepEori2oriVcat promoterCmRlambda t0 terminatoryeGFPPLtetOlacIPLtetO-1 promotertrc promoterlac operatorTetRPtrc-2mCherryrrnB T1 terminator

pBAC-SC vector Sequence

LOCUS       40924_5539       10512 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBAC-SC, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10512)
  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 10512)
  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 10512)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10512)
  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..10512
                     /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(6455..6549)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6612..7325)
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
     regulatory      complement(7377..7450)
                     /label=PLtetO
                     /note="PLtetO"
                     /regulatory_class="promoter"
     promoter        complement(7377..7450)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"
     misc_feature    7396
                     /label=transcription start
                     /note="transcription start"
     protein_bind    7407..7425
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7432..7450
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             complement(7496..8575)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(8612..8685)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator
                     sites (Lutz and Bujard, 1997)"
     promoter        8717..8746
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     prim_transcript 8752
                     /label=transcription start
                     /note="transcription start"
     protein_bind    8754..8770
                     /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)."
     CDS             8817..9437
                     /label=TetR
                     /note="tetracycline repressor TetR"
     regulatory      9483..9557
                     /label=Ptrc-2
                     /note="Ptrc-2"
                     /regulatory_class="promoter"
     promoter        9496..9525
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    9533..9549
                     /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 9533
                     /label=transcription start
                     /note="transcription start"
     CDS             9600..10307
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     terminator      10370..10456
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"

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