LOCUS       Exported                3109 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pNS2Trc, complete sequence.
ACCESSION   EU600162
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3109)
  AUTHORS   Gillaspie D, Perkins I, Larsen K, McCord A, Pangonis S, Sweger D, 
            Seleem MN, Sriranganathan N, Anderson BE.
  TITLE     Plasmid-based system for high-level gene expression and antisense 
            gene knockdown in Bartonella henselae
  JOURNAL   Appl. Environ. Microbiol. 75 (16), 5434-5436 (2009)
  PUBMED    19542333
REFERENCE   2  (bases 1 to 3109)
  AUTHORS   Seleem MN, Anderson B, Sriranganathan N.
  TITLE     Improved Expression Vectors for Bartonella
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 3109)
  AUTHORS   Seleem MN, Anderson B, Sriranganathan N.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-MAR-2008) ICTAS, Virginia Tech, 1410 Prices Fork Rd, 
            Blacksburg, VA 24061, USA
REFERENCE   4  (bases 1 to 3109)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3109
                     /organism="Expression vector pNS2Trc"
                     /mol_type="other DNA"
                     /db_xref="taxon:520152"
     promoter        23..52
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    60..76
                     /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)."
     misc_feature    93..162
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region 
                     (Berg et al., 1989)"
     CDS             213..239
                     /codon_start=1
                     /product="synthetic cistron containing a ribosome binding 
                     site (Shine-Dalgarno sequence), for enhancing the bacterial
                     expression of a downstream cistron (Schoner, 1997)"
                     /label=minicistron
                     /note="This first cistron should terminate 3 bp upstream of
                     the ATG for the second cistron."
                     /translation="MYRLNKEE"
     CDS             246..263
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1454..2116)
                     /codon_start=1
                     /gene="rep"
                     /product="Rep"
                     /label=rep
                     /note="required for plasmid replication"
                     /protein_id="ACC63221.1"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     gene            complement(1454..2116)
                     /gene="rep"
                     /label=rep
     rep_origin      2117..2886
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"