LOCUS       Exported                6207 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pNIT-1, complete sequence.
ACCESSION   FJ173069
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6207)
  AUTHORS   Pandey AK, Raman S, Proff R, Joshi S, Kang CM, Rubin EJ, Husson RN, 
            Sassetti CM.
  TITLE     Nitrile-inducible gene expression in mycobacteria
  JOURNAL   Tuberculosis (Edinb) 89 (1), 12-16 (2009)
  PUBMED    18801704
REFERENCE   2  (bases 1 to 6207)
  AUTHORS   Pandey AK, Proff R, Joshi S, Sassetti CM.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-SEP-2008) Molecular Genetics and Microbiology, 
            University of Massachusetts Medical School, 55 Lake Avenue North. 
            S6-141, Worcester, MA 01655, USA
REFERENCE   3  (bases 1 to 6207)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6207
                     /organism="Expression vector pNIT-1"
                     /mol_type="other DNA"
                     /db_xref="taxon:562926"
     CDS             141..956
                     /codon_start=1
                     /gene="APH"
                     /product="kanamycin 3-phosphotransferase"
                     /label=APH
                     /note="confers resistance to kanamycin"
                     /protein_id="ACI22355.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            141..956
                     /gene="APH"
                     /label=APH
     rep_origin      1265..2030
                     /label=pUC E. coli origin of replication
                     /note="pUC E. coli origin of replication"
     rep_origin      1288..1876
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      2035..3930
                     /label=pAL5000 mycobacterial origin of replication
                     /note="pAL5000 mycobacterial origin of replication"
     terminator      3974..4021
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      4059..4173
                     /regulatory_class="promoter"
                     /note="PnitA"
     misc_feature    4174..4222
                     /label=multiple cloning site
                     /note="multiple cloning site"
     regulatory      4223..4269
                     /regulatory_class="terminator"
                     /note="fd terminator"
     terminator      4229..4268
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     regulatory      4276..4390
                     /regulatory_class="promoter"
                     /note="PnitA"
     CDS             4391..5350
                     /codon_start=1
                     /gene="nitR"
                     /product="nitrile-inducible regulatory protein"
                     /label=nitR
                     /protein_id="ACI22356.1"
                     /translation="MNTFFSSDQVSAPDRVALWHDVICRSYVPLNITLTSEQPFIGTVS
                     TGNLGTVRIATSSSLPQQITRTRRLIRQDEREYLMVGVQSAGHALVQQHGRTARVGRGG
                     LVFWDTRHPYDILFPTDWRMSVFQFPRYSFGFTEDFIGRMTAVNVGGDRGIGRVVSSFM
                     TSINDATDAGDLAEVASLHNSAVDLLSAAIRTELADQAAASDGLLECVLAYIRQNLADP
                     NLCASQIAAEHNVSVRTLHRLFSATGQGVAEHIRNLRLERIKTELADPTSRRYTISALA
                     RKWGFLDPSTFSRAFKDAYGITAREWAASASASPTEVS"
     gene            4391..5350
                     /gene="nitR"
                     /label=nitR
     terminator      5388..5415
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      5507..5593
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      5531..5598
                     /regulatory_class="terminator"
                     /note="rrn terminator"