LOCUS       Exported                5313 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pHIGH100, complete sequence.
ACCESSION   EF216316
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5313)
  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 5313)
  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 5313)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5313
                     /organism="Shuttle vector pHIGH100"
                     /mol_type="other DNA"
                     /note="high copy number mycobacterial-Escherichia coli 
                     shuttle vector"
                     /db_xref="taxon:423235"
     rep_origin      205..992
                     /note="derived from Escherichia coli; OriE"
     rep_origin      250..838
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    1126..1147
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1162..1192
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1200..1216
                     /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     1224..1240
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1328..1344)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1374..3958
                     /note="mycobacterial origin; OriM"
     CDS             complement(4196..5194)
                     /codon_start=1
                     /gene="hygA"
                     /product="hygromycin-B kinase"
                     /label=hygA
                     /note="phosphotransferase; hygromycin resistance; similar 
                     to sequence in GenBank Accession number CAF31839"
                     /protein_id="ABN14087.1"
                     /translation="MTQESLLLLDRIDSDDSYASLRNDQEFWEPLARRALEELGLPVPP
                     VLRVPGESTNPVLVGEPDPVIKLFGEHWCGPESLASESEAYAVLADAPVPVPRLLGRGE
                     LRPGTGAWPWPYLVMSRMTGTTWRSAMDGTTDRNALLALARELGRVLGRLHRVPLTGNT
                     VLTPHSEVFPELLRERRAATVEDHRGWGYLSPRLLDRLEDWLPDVDTLLAGREPRFVHG
                     DLHGTNIFVDLAATEVTGIVDFTDVYAGDSRYSLVQLHLNAFRGDREILAALLDGAQWK
                     RTEDFARELLAFTFLHDFEVFEETPLDLSGFTDPEELAQFLWGPPDTAPGA"
     gene            complement(4196..5194)
                     /gene="hygA"
                     /label=hygA