LOCUS       Exported                8072 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFUN, complete sequence.
ACCESSION   AF038666
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8072)
  AUTHORS   Poquet I, Ehrlich SD, Gruss A.
  TITLE     An export-specific reporter designed for gram-positive bacteria: 
            application to Lactococcus lactis
  JOURNAL   J. Bacteriol. 180 (7), 1904-1912 (1998)
  PUBMED    9537391
REFERENCE   2  (bases 1 to 8072)
  AUTHORS   Poquet I, Gruss A.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-DEC-1997) Laboratoire de Genetique Appliquee-URLGA, 
            Institut National de la Recherche Agronomique, CRJ, Jouy en Josas 
            78352, France
REFERENCE   3  (bases 1 to 8072)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8072
                     /organism="Cloning vector pFUN"
                     /mol_type="genomic DNA"
                     /note="plasmid vector that creates translational fusions 
                     with a nuclease reporter without a signal peptide"
                     /db_xref="taxon:74538"
     misc_feature    1..4580
                     /note="pIL252 moiety; similar to sequence deposited under 
                     GenBank Accession Number AF039139"
     misc_feature    4581..7472
                     /label=similar to BlueScript
                     /note="similar to BlueScript"
     promoter        complement(4599..4617)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4624..4640)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(4781..5236)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5263..5367
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5368..6228
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6399..6987
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    7275..7296
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7311..7341
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7349..7365
                     /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)."
     primer_bind     7373..7389
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7410..7428
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     7458..7474
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      7473..7507
                     /regulatory_class="terminator"
                     /note="trpA terminator"
     misc_feature    7512..7547
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             7543..8010
                     /codon_start=1
                     /gene="delta-nuc"
                     /product="deltaSP-Nuc"
                     /label=delta-nuc
                     /note="nuclease from Staphylococcus aureus devoid of its 
                     signal peptide and the thirteen amino acids at the 
                     N-terminus of mature protein"
                     /protein_id="AAC14660.1"
                     /translation="DPTVYSATSTKKLHKEPATLIKAIDGDTVKLMYKGQPMTFRLLLV
                     DTPETKHPKKGVEKYGPEASAFTKKMVENAKKIEVEFDKGQRTDKYGRGLAYIYADGKM
                     VNEALVRQGLAKVAYVYKPNNTHEQHLRKSEAQAKKEKLNIWSEDNADSGQ"
     gene            7543..8010
                     /gene="delta-nuc"
                     /label=delta-nuc
                     /note="truncated nuc gene potentially encoding deltaSP-Nuc 
                     ORF"
     primer_bind     complement(8061..5)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"