LOCUS       Exported                6984 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pDL276, complete sequence.
ACCESSION   AF216803
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6984)
  AUTHORS   Dunny GM, Lee LN, LeBlanc DJ.
  TITLE     Improved electroporation and cloning vector system for gram-positive
            bacteria
  JOURNAL   Appl. Environ. Microbiol. 57 (4), 1194-1201 (1991)
  PUBMED    1905518
REFERENCE   2  (bases 1 to 6984)
  AUTHORS   Dunny GM, Bae T.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-DEC-1999) Microbiology, University of Minnesota, Box 
            196 UMHC, 1460 Mayo Building, 420 Delaware St. SE, Minneapolis, MN 
            55455, USA
REFERENCE   3  (bases 1 to 6984)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6984
                     /organism="Shuttle vector pDL276"
                     /mol_type="genomic DNA"
                     /label=Gram positive-Gram negative shuttle vector
                     /note="Gram positive-Gram negative shuttle vector"
                     /db_xref="taxon:111474"
     rep_origin      complement(237..825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             1746..2540
                     /codon_start=1
                     /product="aminoglycoside transferase"
                     /label=aminoglycoside transferase
                     /protein_id="AAG13816.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     terminator      5406..5433
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      5525..5611
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5904..5931
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      6023..6109
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(6275..6673)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGECRELFTTPRIGANQFLRRTVNAQTNPWQ
                     NISIASAISSSRTRRISAVLADERRFSA"
     primer_bind     6583..6599
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    6600..6656
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(6669..6685)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6693..6709
                     /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)."
     promoter        complement(6717..6747)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6762..6783
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."