LOCUS       Exported                4444 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pMWO-034, complete sequence.
ACCESSION   JQ283944
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4444)
  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 4444)
  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 4444)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: DNASTAR Lasergene SeqMan Pro v. Version:
                                     7.2.1 (1) Intel
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4444
                     /organism="Expression vector pMWO-034"
                     /mol_type="other DNA"
                     /db_xref="taxon:1166514"
     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    335..2280
                     /label=region containing pSC101 origin of replication
                     /note="region containing pSC101 origin of replication"
     primer_bind     complement(398..420)
                     /note="MWO-rv-1; reverse sequencing primer"
     CDS             complement(736..1686)
                     /codon_start=1
                     /gene="repA"
                     /product="replication initiation protein"
                     /label=repA
                     /protein_id="AFF57847.1"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     gene            complement(736..1686)
                     /gene="repA"
                     /label=repA
     rep_origin      1734..1956
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      2360..2446
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      2400..2443
                     /regulatory_class="terminator"
                     /note="rrnB T1"
     CDS             complement(2497..3120)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor protein class B"
                     /label=tetR
                     /note="from transposon Tn10"
                     /protein_id="AFF57848.1"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNKAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     gene            complement(2497..3120)
                     /gene="tetR"
                     /label=tetR
     terminator      3373..3459
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      3413..3456
                     /regulatory_class="terminator"
                     /note="rrnB T1"
     CDS             complement(3548..4363)
                     /codon_start=1
                     /gene="aphA1"
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=aphA1
                     /protein_id="AFF57849.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(3548..4363)
                     /gene="aphA1"
                     /label=aphA1