LOCUS       Exported               10706 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHR28A10, complete sequence.
ACCESSION   AY237650
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10706
                     /organism="Cloning vector pHR28A10"
                     /mol_type="other DNA"
                     /label=broad-host-range vector
                     /note="broad-host-range vector"
                     /db_xref="taxon:226579"
     misc_feature    1..2642
                     /note="mob and rep regions derived from broad-host-range 
                     vector pBBR1-GFP deposited in GenBank Accession Number 
                     AF110459"
     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..2455
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     CDS             complement(3044..3835)
                     /codon_start=1
                     /gene="aadA"
                     /product="streptomycin and spectinomycin 
                     adenylyltransferase"
                     /label=aadA
                     /note="derived from pMC110"
                     /protein_id="AAO92242.1"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     gene            complement(3044..3835)
                     /gene="aadA"
                     /label=aadA
     regulatory      4744..4776
                     /regulatory_class="promoter"
                     /note="ptac"
     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
                     /regulatory_class="terminator"
                     /note="trpA"
     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"
     gene            complement(6937..8088)
                     /gene="lacIQ"
                     /label=lacIQ
                     /note="promoterless lacIQ gene derived from pSPORT-2"
     regulatory      complement(6937..7005)
                     /regulatory_class="terminator"
                     /gene="lacIQ"
     protein_bind    6972..6993
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(7006..8088)
                     /codon_start=1
                     /gene="lacIQ"
                     /product="LacIQ"
                     /label=lacIQ
                     /note="plac and ptac promoters repressor; expression 
                     regulated by pnifH"
                     /protein_id="AAO92244.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     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)
                     /regulatory_class="promoter"
                     /note="Sinorhizobium meliloti nifH -24/-12 type promoter 
                     derived from pMCK28"
     misc_feature    complement(8761..9053)
                     /label=derived from pMCK28
                     /note="derived from pMCK28"
     CDS             complement(9635..10495)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="derived from pBBR1-GFP deposited in GenBank 
                     Accession Number AF110459"
                     /protein_id="AAO92246.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(9635..10495)
                     /gene="bla"
                     /label=bla