LOCUS       40924_24907        6762 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHR3-km, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6762)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Biotechnol."; date: "2002"; volume: "97"; issue: "3"; pages: 
            "243-252"
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..6762
                     /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
                     /codon_start=1
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     primer_bind     2614..2630
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2985..3776)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    complement(3813..4311)
                     /label=derived from pJQ200KS
                     /note="derived from pJQ200KS"
     promoter        complement(4314..4342)
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    4851..4872
                     /label=CAP binding site
                     /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
                     /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"
     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(5694..6551)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg
       61 cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt
      121 ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg
      181 cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg
      241 cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca
      301 gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt
      361 tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg
      421 tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt
      481 acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg
      541 ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct
      601 tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg
      661 ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc
      721 ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca
      781 tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg
      841 cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg
      901 cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc
      961 ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc
     1021 gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga
     1081 tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc
     1141 ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt
     1201 ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg
     1261 acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac
     1321 gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaaggcgaag cccgcccgcc
     1381 tgccccccga gcctcacggc ggcgagtgcg ggggttccaa gggggcagcg ccaccttggg
     1441 caaggccgaa ggccgcgcag tcgatcaaca agccccggag gggccacttt ttgccggagg
     1501 gggagccgcg ccgaaggcgt gggggaaccc cgcaggggtg cccttctttg ggcaccaaag
     1561 aactagatat agggcgaaat gcgaaagact taaaaatcaa caacttaaaa aaggggggta
     1621 cgcaacagct cattgcggca ccccccgcaa tagctcattg cgtaggttaa agaaaatctg
     1681 taattgactg ccacttttac gcaacgcata attgttgtcg cgctgccgaa aagttgcagc
     1741 tgattgcgca tggtgccgca accgtgcggc accctaccgc atggagataa gcatggccac
     1801 gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga
     1861 acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct
     1921 gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac
     1981 actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt
     2041 ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt
     2101 ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag
     2161 tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct
     2221 gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga
     2281 gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac
     2341 ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct
     2401 ggacgatggc gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcactt
     2461 gggttgcgca gcaacccgta agtgcgctgt tccagactat cggctgtagc cgcctcgccg
     2521 ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga
     2581 cctgaatgga agccccaccg cggtggcggc cgctctagaa ctagtggatc ctctagagtc
     2641 gaccaaagcg gccatcgtgc ctccccactc ctgcagttcg ggggcatgga tgcgcggata
     2701 gccgctgctg gtttcctgga tgccgacgga tttgcactgc cggtagaact ccgcgaggtc
     2761 gtccagcctc aggcagcagc tgaaccaact cgcgagggga tcgagcccgg ggtgggcgaa
     2821 gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat
     2881 tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt
     2941 gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga
     3001 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
     3061 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
     3121 tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
     3181 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg
     3241 ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
     3301 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
     3361 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
     3421 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
     3481 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
     3541 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt
     3601 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
     3661 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
     3721 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga
     3781 aacgatcctc atcctgtctc ttgatcagat ctcactacgc ggctgctcaa acctgggcag
     3841 aacgtaagcc gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa
     3901 tgtcttacta cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt
     3961 ggctacgtct ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg
     4021 gtcagggccg agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt
     4081 agggcgactg ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata
     4141 acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact
     4201 gtacaaaaaa acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc
     4261 gttcggtcaa ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac
     4321 gaaccgaaca ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc
     4381 acccggcaac cttgggcagc agcgaagtcg aggcatttct gtcctggctg gcgaacgagc
     4441 gcaaggtttc ggtctccacg catcgtcagg cattggcggc cttgctgttc ttctacggca
     4501 aggtgctgtg cacggatctg ccctggcttc aggagatcgg aagacctcgg ccgtcgcggc
     4561 gcttgccggt ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg
     4621 agcatcgttt gttcgcccag cttctgtatg gaacgggcat gcggatcagt gagggtttgc
     4681 aactgcgggt caaggatctg gatttcgatc acggcacgat catcgtgcgg gagggcaagg
     4741 gctccaagga tcgggccttg atgttacccg agagcttggc acccacgctg cgcgagcccc
     4801 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     4861 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     4921 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     4981 cgcgcaatta accctcacta aagggaacaa aagctgggta ccgggccccc cctcgaggtc
     5041 gacggtatcg ataagcttga tatcgaattc ctgcagcccg ggggatctgg taccgggccc
     5101 cccctcgagg gggggcccgt ttctcaaagc tcacgctgta ggtatctcag ttcggtgtag
     5161 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5221 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5281 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5341 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5401 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5461 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5521 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5581 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5641 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5701 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5761 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5821 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5881 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5941 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6001 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6061 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6121 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6181 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6241 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6301 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6361 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6421 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6481 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6541 tgaatactca tactcttcct ttttcaatca ggtggcactt ttcggggaaa tgtgcgcgcc
     6601 cgcgttcctg ctggcgctgg gcctgtttct ggcgctggac ttcccgctgt tccgtcagca
     6661 gcttttcgcc cacggccttg atgatcgcgg cggccttggc ctgcatatcc cgattcaacg
     6721 gccccagggc gtccagaacg ggcttcaggc gctcccgaag gt
//
