LOCUS       Exported                3329 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pMWO-077, complete sequence.
ACCESSION   JQ283949
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3329)
  AUTHORS   Obrist MW, Miller VL.
  TITLE     Low copy expression vectors for use in Yersinia sp. and related 
            organisms
  JOURNAL   Plasmid 68 (1), 33-42 (2012)
  PUBMED    22445322
REFERENCE   2  (bases 1 to 3329)
  AUTHORS   Obrist MW, Miller VL.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-DEC-2011) Microbiology and Immunology, University of 
            North Carolina at Chapel Hill, 116 Manning Drive, 505 Mary Ellen 
            Jones, CB# 7290, Chapel Hill, NC 27599, USA
REFERENCE   3  (bases 1 to 3329)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: DNASTAR Lasergene SeqMan Pro v. 7.2.1 (1)
                                     Intel
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3329
                     /organism="Expression vector pMWO-077"
                     /mol_type="other DNA"
                     /db_xref="taxon:1166519"
     primer_bind     60..77
                     /note="MWO-fw; forward sequencing primer"
     promoter        75..148
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    75..93
                     /label=tet operator 2 binding site
                     /bound_moiety="tet operator 2"
                     /note="operator 2 from Tn10; tet repressor binding site"
     regulatory      94..99
                     /regulatory_class="minus_35_signal"
     protein_bind    100..118
                     /label=tet operator 2 binding site
                     /bound_moiety="tet operator 2"
                     /note="operator 2 from Tn10; tet repressor binding site"
     regulatory      117..122
                     /regulatory_class="minus_10_signal"
     misc_feature    155..297
                     /label=multiple cloning site from pBluescript II KS+
                     /note="multiple cloning site from pBluescript II KS+"
     misc_feature    155..262
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     172..188
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(222..238)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(271..289)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    298..1202
                     /label=region containing p15A origin of replication
                     /note="region containing p15A origin of replication"
     primer_bind     complement(384..405)
                     /note="MWO-rv-2; reverse sequencing primer"
     rep_origin      complement(557..1102)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     terminator      1245..1331
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      1285..1328
                     /regulatory_class="terminator"
                     /note="rrnB T1"
     CDS             complement(1382..2005)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor protein class B"
                     /label=tetR
                     /note="from transposon Tn10"
                     /protein_id="AFF57860.1"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNKAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     gene            complement(1382..2005)
                     /gene="tetR"
                     /label=tetR
     terminator      2258..2344
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      2298..2341
                     /regulatory_class="terminator"
                     /note="rrnB T1"
     CDS             complement(2433..3248)
                     /codon_start=1
                     /gene="aphA1"
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=aphA1
                     /protein_id="AFF57861.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(2433..3248)
                     /gene="aphA1"
                     /label=aphA1