LOCUS       Exported                9037 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHRE1-km, complete sequence.
ACCESSION   AY286001
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9037)
  AUTHORS   Ramos HJO., Soares-Ramos JRL., Souza EM, Pedrosa FO.
  TITLE     Broad-host-range cloning vector pHRE1-km for promoter trapping
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9037)
  AUTHORS   Ramos HJO., Soares-Ramos JRL., Souza EM, Pedrosa FO.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-APR-2003) Department of Biochemistry and Molecular 
            Biology, Universidade Federal do Parana-UFPR, Centro Politecnico, 
            Curitiba, Parana 81531-990, Brazil
REFERENCE   3  (bases 1 to 9037)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9037
                     /organism="Cloning vector pHRE1-km"
                     /focus
                     /mol_type="other DNA"
                     /note="broad host-range vector used to make transcriptional
                     fusions to both gfp and uidA"
                     /db_xref="taxon:229474"
     source          6591..2642
                     /organism="Cloning vector pBBR1-GFP"
                     /mol_type="other DNA"
                     /note="from sequence found in GenBank Accession Number 
                     AF110459"
                     /db_xref="taxon:92689"
     misc_feature    1..2642
                     /label=mob and rep region
                     /note="mob and rep region"
     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"
                     /product="aminoglycoside-3-O-phosphotransferase"
                     /label=aph
                     /note="NPTII; from vector pWM6"
                     /protein_id="AAP31912.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(2982..3776)
                     /gene="aph"
                     /label=aph
     regulatory      complement(3865..3914)
                     /regulatory_class="promoter"
                     /note="promoter region of neomycin and kanamycin resistance
                     aph (NPTII) gene"
     CDS             complement(4706..6517)
                     /codon_start=1
                     /gene="uidA"
                     /product="beta-glucuronidase"
                     /label=uidA
                     /protein_id="AAP31913.1"
                     /translation="MVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     gene            complement(4706..6517)
                     /gene="uidA"
                     /label=uidA
                     /note="promoterless gene from pWM6"
     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 fluorescent protein"
                     /label=gfp
                     /note="modified form mut3 from Aequorea victoria GFP found 
                     in GenBank Accession number M62653"
                     /protein_id="AAP31914.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     gene            complement(6610..7326)
                     /gene="gfp"
                     /label=gfp
                     /note="promoterless gene"
     CDS             complement(7966..8826)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AAP31915.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(7966..8826)
                     /gene="bla"
                     /label=bla