LOCUS       Exported                3751 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMC1-EPS-RtGFP, complete sequence.
ACCESSION   KC989925
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3751)
  AUTHORS   Madhaiyan M, Peng N, Ngoh ST, Cheng H, Cai L, Fu L, Reddy C, Hong Y,
            Ji L.
  TITLE     Synthetic Vector pMC1-EPS-RtGFP
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3751)
  AUTHORS   Madhaiyan M, Peng N, Ngoh ST, Cheng H, Cai L, Fu L, Reddy C, Hong Y,
            Ji L.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-APR-2013) Biomaterials & Biocatalysts Group, Temasek 
            Life Sciences Laboratory, 1 Research Link, National University of 
            Singapore, Singapore 117604, Singapore
REFERENCE   3  (bases 1 to 3751)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3751
                     /organism="Cloning vector pMC1-EPS-RtGFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1389800"
     promoter        complement(6..50)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    14..30
                     /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)."
     regulatory      complement(54..3725)
                     /regulatory_class="promoter"
                     /note="tac promoter"
     regulatory      58..753
                     /regulatory_class="promoter"
                     /note="eps promoter"
     CDS             754..1473
                     /codon_start=1
                     /product="eGFP"
                     /label=eGFP
                     /protein_id="AGV01071.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     rep_origin      complement(1866..2534)
                     /direction=LEFT
                     /label=ColE1
                     /note="ColE1"
     rep_origin      1889..2477
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(2800..2833)
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     polyA_signal    2926..3047
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3055..3696)
                     /codon_start=1
                     /product="CatII"
                     /label=CatII
                     /protein_id="AGV01070.1"
                     /translation="MNFTRIDLNTWNRREHFALYRQQIKCGFSLTTKLDITALRTALAE
                     TGYKFYPLMIYLISRAVNQFPEFRMALKDNELIYWDQSDPVFTVFHKETETFSALSCRY
                     FPDLSEFMAGYNAVTAEYQHDTRLFPQGNLPENHLNISSLPWVSFDGFNLNITGNDDYF
                     APVFTMAKFQQEGDRVLLPVSVQVHHAVCDGFHAARFINTLQLMCDNILK"
     RBS             3703..3714
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    3733..3749
                     /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)."