LOCUS       Exported                3562 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pIJ12738, complete sequence.
ACCESSION   KU523901
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3562)
  AUTHORS   Fernandez-Martinez LT, Bibb MJ.
  TITLE     Use of the meganuclease I-SceI of Saccharomyces cerevisiae to select
            for gene deletions in actinomycetes
  JOURNAL   Sci Rep 4, 7100 (2014)
  PUBMED    25403842
REFERENCE   2  (bases 1 to 3562)
  AUTHORS   Fernandez-Martinez LT, Bibb MJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-JAN-2016) Biology, Edge Hill University, St Helens 
            Road, Ormskirk L39 4QP, UK
REFERENCE   3  (bases 1 to 3562)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3562
                     /organism="Cloning vector pIJ12738"
                     /mol_type="other DNA"
                     /db_xref="taxon:1825501"
     misc_feature    complement(163..479)
                     /label=fragment of lacZ alpha peptide
                     /note="fragment of lacZ alpha peptide"
     primer_bind     423..439
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     490..506
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(609..622)
                     /label=fragment of lacZ alpha peptide
                     /note="fragment of lacZ alpha peptide"
     primer_bind     complement(623..639)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    647..663
                     /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(671..701)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    716..737
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1025..1613)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             1817..2620
                     /codon_start=1
                     /gene="aacIV"
                     /product="apramycin resistance protein"
                     /label=aacIV
                     /protein_id="AMV91697.1"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     gene            1817..2620
                     /gene="aacIV"
                     /label=aacIV
     CDS             1817..2617
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     rep_origin      3038..3149
     oriT            3040..3149
                     /note="incP origin of transfer"