LOCUS       Exported                6762 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHR3-km, complete sequence.
ACCESSION   AY237649
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6762)
  AUTHORS   Ramos HJ, Roncato-Maccari LD, Souza EM, Soares-Ramos JR, Hungria M, 
            Pedrosa FO.
  TITLE     Monitoring Azospirillum-wheat interactions using the gfp and gusA 
            genes constitutively expressed from a new broad-host range vector
  JOURNAL   J. Biotechnol. 97 (3), 243-252 (2002)
  PUBMED    12084480
REFERENCE   2  (bases 1 to 6762)
  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 6762)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6762
                     /organism="Cloning vector pHR3-km"
                     /mol_type="genomic DNA"
                     /label=broad-host-range vector
                     /note="broad-host-range vector"
                     /db_xref="taxon:226578"
     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"
     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="kanamycin resistance gene; expression regulated by 
                     aacC1 promoter"
                     /protein_id="AAO92240.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(2982..3776)
                     /gene="aph"
                     /gene_synonym="NptII"
                     /label=aph
                     /note="derived from Cloning vector pWM6"
     misc_feature    complement(3813..4311)
                     /label=derived from pJQ200KS
                     /note="derived from pJQ200KS"
     regulatory      complement(4314..4342)
                     /regulatory_class="promoter"
                     /note="contains -35 and -10 promoter regions derived from 
                     gentamycin resistance aacC1 gene in pJQ200KS"
     promoter        complement(4314..4342)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    4851..4872
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4887..4917
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4925..4941
                     /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     4949..4965
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4986..5004
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    complement(5017..5088)
                     /note="multiple cloning site; derived from pBluescript-II"
     primer_bind     5034..5050
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(5087..5690)
                     /label=derived from pJQ200KS
                     /note="derived from pJQ200KS"
     CDS             complement(5691..6551)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="derived from pBBR1-GFP deposited in GenBank 
                     Accession Number AF110459"
                     /protein_id="AAO92241.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(5691..6551)
                     /gene="bla"
                     /label=bla