pHR28A10 vector (V005508)

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LOCUS       40924_24902       10706 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHR28A10, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10706)
  AUTHORS   Ramos HJ, Souza EM, Soares-Ramos JR, Pedrosa FO.
  TITLE     A new system to control the barnase expression by a NifA-dependent 
            promoter
  JOURNAL   J. Biotechnol. 118 (1), 9-16 (2005)
  PUBMED    15908028
REFERENCE   2  (bases 1 to 10706)
  AUTHORS   Ramos HJO., Soares-Ramos JRL., Souza EM, Pedrosa FO.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-FEB-2003) Department of Biochemistry and Molecular 
            Biology, Universidade Federal do Parana - UFPR, Centro Politecnico, 
            Curitiba, Parana 81531-990, Brazil
REFERENCE   3  (bases 1 to 10706)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10706)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Biotechnol."; date: "2005"; volume: "118"; issue: "1"; pages: "9-16"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (14-FEB-2003) Department of Biochemistry and Molecular Biology, 
            Universidade Federal do Parana - UFPR, Centro Politecnico, Curitiba,
            Parana 81531-990, Brazil"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1023..1792
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1793..2452
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     CDS             complement(3047..3835)
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     regulatory      4744..4776
                     /label=ptac
                     /note="ptac"
                     /regulatory_class="promoter"
     promoter        4746..4773
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    4781..4797
                     /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             4863..6671
                     /codon_start=1
                     /gene="uidA"
                     /product="beta-glucuronidase"
                     /label=uidA
                     /note="expression regulated by ptac; derived from pMC110"
                     /protein_id="AAO92243.1"
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGQQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQVHGNISPLAEATRKLDPTRPITCVNVMFCDAHTDTISDL
                     FDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMYT
                     DMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRKP
                     KSAAFLLQKRWTGMNFGEKPQQGGKQ"
     gene            4863..6671
                     /gene="uidA"
                     /label=uidA
     regulatory      6735..6763
                     /label=trpA
                     /note="trpA"
                     /regulatory_class="terminator"
     primer_bind     6798..6814
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(6842..6860)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6879..6895)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6903..6919
                     /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)."
     promoter        complement(6927..6957)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6972..6993)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(7009..8088)
                     /label=lacI
                     /note="lac repressor"
     CDS             complement(8165..8437)
                     /codon_start=1
                     /gene="barstar"
                     /product="barnase inhibitor"
                     /label=barstar
                     /note="expression regulated by pnifH"
                     /protein_id="AAO92245.1"
                     /translation="MKKAVINGEQIRSISDLHQTLKKELALPEYYGENLDALWDCLTGW
                     VEYPLVLEWRQFEQSKQLTENGAESVLQVFREAKAEGCDITIILS"
     gene            complement(8165..8437)
                     /gene="barstar"
                     /label=barstar
                     /note="promoterless barstar gene derived from pMT316
                     deposited in GenBank Accession Number X12871"
     regulatory      complement(8608..8757)
                     /note="Sinorhizobium meliloti nifH -24/-12 type promoter
                     derived from pMCK28"
                     /regulatory_class="promoter"
     misc_feature    complement(8761..9053)
                     /label=derived from pMCK28
                     /note="derived from pMCK28"
     CDS             complement(9638..10495)
                     /label=AmpR
                     /note="beta-lactamase"