pBAC-L6-PT3-T4Ei vector (V009335)

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"