LOCUS       Exported                4892 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pWB101, complete sequence.
ACCESSION   EF216318
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4892)
  AUTHORS   Bourn WR, Jansen Y, Stutz H, Warren RM, Williamson AL, van Helden 
            PD.
  TITLE     Creation and characterisation of a high-copy-number version of the 
            pAL5000 mycobacterial replicon
  JOURNAL   Tuberculosis (Edinb) 87 (6), 481-488 (2007)
  PUBMED    17888739
REFERENCE   2  (bases 1 to 4892)
  AUTHORS   Bourn WR.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-JAN-2007) Medical Virology and Institute of Infectious
            Disease and Molecular Medicine, University of Cape Town, Anzio Road 
            Observatory, Cape Town 7925, South Africa
REFERENCE   3  (bases 1 to 4892)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4892
                     /organism="Shuttle vector pWB101"
                     /mol_type="other DNA"
                     /label=mycobacterial-Escherichia coli shuttle vector
                     /note="mycobacterial-Escherichia coli shuttle vector"
                     /db_xref="taxon:423239"
     CDS             120..935
                     /codon_start=1
                     /gene="aph"
                     /product="aminoglycoside-3-phosphotransferase"
                     /label=aph
                     /note="kanamycin resistance"
                     /protein_id="ABN14089.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            120..935
                     /gene="aph"
                     /label=aph
     rep_origin      1222..2009
                     /note="derived from Escherichia coli; OriE"
     rep_origin      1267..1855
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      2014..3909
                     /note="mycobacterial origin; OriM"
     regulatory      3936..4022
                     /regulatory_class="promoter"
                     /note="BCG hsp60"
     regulatory      4046..4052
                     /regulatory_class="ribosome_binding_site"
                     /note="strong synthetic Shine Delgarno sequence; RBSA"
     CDS             4062..4778
                     /codon_start=1
                     /gene="gfp"
                     /product="green fluorescent protein"
                     /label=gfp
                     /note="fluorescent reporter"
                     /protein_id="ABN14090.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     gene            4062..4778
                     /gene="gfp"
                     /label=gfp
     terminator      4810..4856
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      4813..4856
                     /regulatory_class="terminator"
                     /note="rrnBT1"