LOCUS       Exported               11973 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHR50, complete sequence.
ACCESSION   AY237648
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11973)
  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   2  (bases 1 to 11973)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11973
                     /organism="Cloning vector pHR50"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:227048"
     misc_feature    1..2642
                     /note="mob and rep regions from broad-host-range vector 
                     pBBR1-GFP deposited in GeneBank 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"
     primer_bind     2614..2630
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2982..3776)
                     /codon_start=1
                     /gene="aph"
                     /gene_synonym="nptII"
                     /product="aminoglycoside-3-O-phosphotransferase"
                     /label=aph
                     /note="NPTII kanamycin resistance gene"
                     /protein_id="AAP04589.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(2982..3776)
                     /gene="aph"
                     /gene_synonym="nptII"
                     /label=aph
                     /note="derived from pWM6"
     CDS             complement(4706..6517)
                     /codon_start=1
                     /gene="uidA"
                     /product="beta-glucuronidase"
                     /label=uidA
                     /protein_id="AAP04590.1"
                     /translation="MVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     gene            complement(4706..6517)
                     /gene="uidA"
                     /label=uidA
                     /note="derived from pWM6; expressed from gentamycin 
                     resistance aacC1 promoter"
     primer_bind     complement(6555..6571)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(6610..7326)
                     /codon_start=1
                     /gene="gfp"
                     /product="green florescent protein"
                     /label=gfp
                     /note="modified form mut3; derived from Aequorea victoria 
                     GFP deposited in GenBank Accession number M62653"
                     /protein_id="AAP04593.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     gene            complement(6610..7326)
                     /gene="gfp"
                     /label=gfp
                     /note="dervied from pBBR1-GFP; expressed from gentamycin 
                     resistance aacC1 promoter"
     CDS             complement(7376..8950)
                     /codon_start=1
                     /gene="nifA"
                     /product="nif genes transcriptional activator"
                     /label=nifA
                     /protein_id="AAP04591.1"
                     /translation="MIHKSDSDTTVRRFDLSQQFTAMQRISVVLSRATEASKTLQEVLS
                     VLHNDAFMQHGMICLYDSQQEILSIEALQQTEDQTLPGSTQIRYRPGEGLVGTVLAQGQ
                     SLVLPRVADDQRFLDRLSLYDYDLPFIAVPLMGPHSRPIGVLAAHAMARQEERLPACTR
                     FLETVANLIAQTIRLMILPTSAAQAPQQSPRIERPRACTPSRGFGLENMVGKSPAMRQI
                     MDIIRQVSRWDTTVLVRGESGTGKELIANAIHHNSPRAAAAFVKFNCAALPDNLLESEL
                     FGHEKGAFTGAVRQRKGRFELADGGTLFLDEIGESSASFQAKLLRILQEGEMERVGGDE
                     TLRVNVRIIAATNRHLEEEVRLGHFREDLYYRLNVMPIALPPLRERQEDIAELAHFLVR
                     KIAHSQGRTLRISDGAIRLLMEYSWPGNVRELENCLERSAVLSESGLIDRDVILFNHRD
                     NPPKALASSGPAEDGWLDNSLDERQRLIAALEKAGWVQAKAARLLGMTPRQVAYRIQIM
                     DITMPRL"
     gene            complement(7376..8950)
                     /gene="nifA"
                     /label=nifA
                     /note="derived from Klebsiella pneumoniae nifA gene of 
                     pNH11; expressed from gentamycin resistance aacC1 promoter"
     misc_feature    8951..10298
                     /label=derived from pJQ200KS
                     /note="derived from pJQ200KS"
     regulatory      complement(9520..9525)
                     /regulatory_class="minus_10_signal"
                     /note="derived from gentamycin resistance aacC1 gene of 
                     pJQ200KS"
     promoter        complement(9525..9553)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     regulatory      complement(9548..9553)
                     /regulatory_class="minus_35_signal"
                     /note="derived from gentamycin resistance aacC1 gene of 
                     pJQ200KS"
     protein_bind    10062..10083
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        10098..10128
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10136..10152
                     /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)."
     primer_bind     10160..10176
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        10197..10215
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     10245..10261
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(10902..11762)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AAP04592.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(10902..11762)
                     /gene="bla"
                     /label=bla
                     /note="derived from pBBR1-GFP"