pKEK1140 vector (V005247)

LOCUS       V005247                 8419 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005247
VERSION     V005247
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8419)
  AUTHORS   Rodriguez SA, Yu JJ, Davis G, Arulanandam BP, Klose KE.
  TITLE     Targeted inactivation of francisella tularensis genes by group II
            introns
  JOURNAL   Appl. Environ. Microbiol. 74 (9), 2619-2626 (2008)
   PUBMED   18310413
REFERENCE   2  (bases 1 to 8419)
  AUTHORS   Rodriguez SA, Petrosino JF, Klose KE.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-FEB-2008) Biology, South Texas Center for Emerging
            Infectious Diseases, The University of Texas at San Antonio, One
            UTSA Circle, San Antonio, TX 78249, USA
REFERENCE   3  (bases 1 to 8419)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8419)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
            Environ. Microbiol."; date: "2008"; volume: "74"; issue: "9"; pages:
            "2619-2626"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (18-FEB-2008) Biology, South Texas Center for Emerging Infectious
            Diseases, The University of Texas at San Antonio, One UTSA Circle,
            San Antonio, TX 78249, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8419
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        142..172
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..195
                     /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)."
     CDS             216..389
                     /label="lacZ-alpha"
                     /note="LacZ-alpha fragment of beta-galactosidase"
     intron          770..1372
                     /note="Lactococcus lactis LtrB group II intron"
     RBS             1405..1427
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1619..3415
                     /gene="ltrA"
                     /label="Group II intron-encoded protein LtrA"
                     /note="Group II intron-encoded protein LtrA from
                     Lactococcus lactis subsp. cremoris (strain MG1363).
                     Accession#: P0A3U1"
     terminator      3610..3696
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3788..3815
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     rep_origin      4036..4581
                     /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."
     misc_feature    complement(4912..6807)
                     /note="corresponds to region within Francisella tularensis
                     subsp. novicida plasmid, pFNL10, which encodes origin of
                     replication"
     CDS             complement(5273..6292)
                     /codon_start=1
                     /gene="repA"
                     /product="temperature-sensitive RepA"
                     /label="repA"
                     /note="contains amino acid mutation, methionine to
                     isoleucine M120I"
                     /protein_id="ACA63832.1"
                     /translation="MKEVKNVNAGKEIAMSNTLVAGKYSLTKEEQNLIFLVASMVKRED
                     KEFHRYKISLSDLEKATGVKHNRVRLKQLMHSIMSKPVWLNKEQTKIANWFAYIEADPK
                     SSALICEFHWSLMPHIIQLQEYFTKAERQLLFSFKSKYSSRLYLLLKSKLGEQAGYANI
                     VDCVLYVDDMINDFDLPKSYSNRYSNFKNKFLLPALEEINQLSDIFVTYDDKDHHRKHG
                     RKITNIKFTVSKVAETEEEFKQKLLDTKNKEDYIPIEASERLREVVLSDELDLDLYYVR
                     SIFRHHHLHDIERVCDDTWRNWDNKKIKDRKRVFIANIKKLEKKPEENHKLPFGFEEV"
     gene            complement(5273..6292)
                     /gene="repA"
                     /label="repA"
     regulatory      7163..7322
                     /note="region encodes promoter of FTN1451 of Francisella
                     tularensis subsp. novicida"
                     /regulatory_class="promoter"
     CDS             7326..8138
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     regulatory      8181..8419
                     /note="region encodes groELp promoter of Francisella
                     tularensis subsp. holarctica strain, LVS"
                     /regulatory_class="promoter"