LOCUS       sequence-325159-        6553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IncQ BHR plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6553)
  AUTHORS   Tapscott T, Guarnieri MT, Henard CA
  TITLE     Development of a CRISPR/Cas9 System for Methylococcus capsulatus In 
            Vivo Gene Editing.
  JOURNAL   Appl Environ Microbiol. 2019 May 16;85(11). pii: AEM.00340-19. doi: 
            10.1128/AEM.00340-19. Print 2019 Jun 1.
  PUBMED    30926729
REFERENCE   2  (bases 1 to 6553)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl 
            Environ Microbiol."; date: "2019-05-16"; volume: "85(11). pii: 
            AEM.00340-19. doi"; pages: " 10.1128/AEM.00340-19. Print 2019 Jun 1"
COMMENT     Sequence Label: pQCH
FEATURES             Location/Qualifiers
     source          1..6553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            1..88
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             1327..2298
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     CDS             2809..3648
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             3635..4486
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     primer_bind     complement(4734..4753)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4745..5560
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             4745..5560
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4818..4837)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5818..6212
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
ORIGIN
        1 gtaggctatc atggaggcac agcggcggca atcccgaccc tactttgtag gggagggcgc
       61 acttaccggt ttctcttcga gaaactggcc taacggccac ccttcgggcg gtgcgctctc
      121 cgagggccat tgcatggagc cgaaaagcaa aagcaacagc gaggcagcat ggcgatttat
      181 caccttacgg cgaaaaccgg cagcaggtcg ggcggccaat cggccagggc caaggccgac
      241 tacatccagc gcgaaggcaa gtatgcccgc gacatggatg aagtcttgca cgccgaatcc
      301 gggcacatgc cggagttcgt cgagcggccc gccgactact gggatgctgc cgacctgtat
      361 gaacgcgcca atgggcggct gttcaaggag gtcgaatttg ccctgccggt cgagctgacc
      421 ctcgaccagc agaaggcgct ggcgtccgag ttcgcccagc acctgaccgg tgccgagcgc
      481 ctgccgtata cgctggccat ccatgccggt ggcggcgaga acccgcactg ccacctgatg
      541 atctccgagc ggatcaatga cggcatcgag cggcccgccg ctcagtggtt caagcggtac
      601 aacggcaaga ccccggagaa gggcggggca cagaagaccg aagcgctcaa gcccaaggca
      661 tggcttgagc agacccgcga ggcatgggcc gaccatgcca accgggcatt agagcgggct
      721 ggccacgacg cccgcattga ccacagaaca cttgaggcgc agggcatcga gcgcctgccc
      781 ggtgttcacc tggggccgaa cgtggtggag atggaaggcc ggggcatccg caccgaccgg
      841 gcagacgtgg ccctgaacat cgacaccgcc aacgcccaga tcatcgactt acaggaatac
      901 cgggaggcaa tagaccatga acgcaatcga cagagtgaag aaatccagag gcatcaacga
      961 gttagcggag cagatcgaac cgctggccca gagcatggcg acactggccg acgaagcccg
     1021 gcaggtcatg agccagaccc agcaggccag cgaggcgcag gcggcggagt ggctgaaagc
     1081 ccagcgccag acaggggcgg catgggtgga gctggccaaa gagttgcggg aggtagccgc
     1141 cgaggtgagc agcgccgcgc agagcgcccg gagcgcgtcg cgggggtggc actggaagct
     1201 atggctaacc gtgatgctgg cttccatgat gcctacggtg gtgctgctga tcgcatcgtt
     1261 gctcttgctc gacctgacgc cactgacaac cgaggacggc tcgatctggc tgcgcttggt
     1321 ggcccgatga agaacgacag gactttgcag gccataggcc gacagctcaa ggccatgggc
     1381 tgtgagcgct tcgatatcgg cgtcagggac gccaccaccg gccagatgat gaaccgggaa
     1441 tggtcagccg ccgaagtgct ccagaacacg ccatggctca agcggatgaa tgcccagggc
     1501 aatgacgtgt atatcaggcc cgccgagcag gagcggcatg gtctggtgct ggtggacgac
     1561 ctcagcgagt ttgacctgga tgacatgaaa gccgagggcc gggagcctgc cctggtagtg
     1621 gaaaccagcc cgaagaacta tcaggcatgg gtcaaggtgg ccgacgccgc aggcggtgaa
     1681 cttcgggggc agattgcccg gacgctggcc agcgagtacg acgccgaccc ggccagcgcc
     1741 gacagccgcc actatggccg cttggcgggc ttcaccaacc gcaaggacaa gcacaccacc
     1801 cgcgccggtt atcagccgtg ggtgctgctg cgtgaatcca agggcaagac cgccaccgct
     1861 ggcccggcgc tggtgcagca ggctggccag cagatcgagc aggcccagcg gcagcaggag
     1921 aaggcccgca ggctggccag cctcgaactg cccgagcggc agcttagccg ccaccggcgc
     1981 acggcgctgg acgagtaccg cagcgagatg gccgggctgg tcaagcgctt cggtgatgac
     2041 ctcagcaagt gcgactttat cgccgcgcag aagctggcca gccggggccg cagtgccgag
     2101 gaaatcggca aggccatggc cgaggccagc ccagcgctgg cagagcgcaa gcccggccac
     2161 gaagcggatt acatcgagcg caccgtcagc aaggtcatgg gtctgcccag cgtccagctt
     2221 gcgcgggccg agctggcacg ggcaccggca ccccgccagc gaggcatgga caggggcggg
     2281 ccagatttca gcatgtagtg cttgcgttgg tactcacgcc tgttatacta tgagtactca
     2341 cgcacagaag ggggttttat ggaatacgaa aaaagcgctt cagggtcggt ctacctgatc
     2401 aaaagtgaca agggctattg gttgcccggt ggctttggtt atacgtcaaa caaggccgag
     2461 gctggccgct tttcagtcgc tgatatggcc agccttaacc ttgacggctg caccttgtcc
     2521 ttgttccgcg aagacaagcc tttcggcccc ggcaagtttc tcggtgactg atatgaaaga
     2581 ccaaaaggac aagcagaccg gcgacctgct ggccagccct gacgctgtac gccaagcgcg
     2641 atatgccgag cgcatgaagg ccaaagggat gcgtcagcgc aagttctggc tgaccgacga
     2701 cgaatacgag gcgctgcgcg agtgcctgga agaactcaga gcggcgcagg gcgggggtag
     2761 tgaccccgcc agcgcctaac caccaactgc ctgcaaagga ggcaatcaat ggctacccat
     2821 aagcctatca atattctgga ggcgttcgca gcagcgccgc caccgctgga ctacgttttg
     2881 cccaacatgg tggccggtac ggtcggggcg ctggtgtcgc ccggtggtgc cggtaaatcc
     2941 atgctggccc tgcaactggc cgcacagatt gcaggcgggc cggatctgct ggaggtgggc
     3001 gaactgccca ccggcccggt gatctacctg cccgccgaag acccgcccac cgccattcat
     3061 caccgcctgc acgcccttgg ggcgcacctc agcgccgagg aacggcaagc cgtggctgac
     3121 ggcctgctga tccagccgct gatcggcagc ctgcccaaca tcatggcccc ggagtggttc
     3181 gacggcctca agcgcgccgc cgagggccgc cgcctgatgg tgctggacac gctgcgccgg
     3241 ttccacatcg aggaagaaaa cgccagcggc cccatggccc aggtcatcgg tcgcatggag
     3301 gccatcgccg ccgataccgg gtgctctatc gtgttcctgc accatgccag caagggcgcg
     3361 gccatgatgg gcgcaggcga ccagcagcag gccagccggg gcagctcggt actggtcgat
     3421 aacatccgct ggcagtccta cctgtcgagc atgaccagcg ccgaggccga ggaatggggt
     3481 gtggacgacg accagcgccg gttcttcgtc cgcttcggtg tgagcaaggc caactatggc
     3541 gcaccgttcg ctgatcggtg gttcaggcgg catgacggcg gggtgctcaa gcccgccgtg
     3601 ctggagaggc agcgcaagag caagggggtg ccccgtggtg aagcctaaga acaagcacag
     3661 cctcagccac gtccggcacg acccggcgca ctgtctggcc cccggcctgt tccgtgccct
     3721 caagcggggc gagcgcaagc gcagcaagct ggacgtgacg tatgactacg gcgacggcaa
     3781 gcggatcgag ttcagcggcc cggagccgct gggcgctgat gatctgcgca tcctgcaagg
     3841 gctggtggcc atggctgggc ctaatggcct agtgcttggc ccggaaccca agaccgaagg
     3901 cggacggcag ctccggctgt tcctggaacc caagtgggag gccgtcaccg ctgatgccat
     3961 ggtggtcaaa ggtagctatc gggcgctggc aaaggaaatc ggggcagagg tcgatagtgg
     4021 tggggcgctc aagcacatac aggactgcat cgagcgcctt tggaaggtat ccatcatcgc
     4081 ccagaatggc cgcaagcggc aggggtttcg gctgctgtcg gagtacgcca gcgacgaggc
     4141 ggacgggcgc ctgtacgtgg ccctgaaccc cttgatcgcg caggccgtca tgggtggcgg
     4201 ccagcatgtg cgcatcagca tggacgaggt gcgggcgctg gacagcgaaa ccgcccgcct
     4261 gctgcaccag cggctgtgtg gctggatcga ccccggcaaa accggcaagg cttccataga
     4321 taccttgtgc ggctatgtct ggccgtcaga ggccagtggt tcgaccatgc gcaagcgccg
     4381 ccagcgggtg cgcgaggcgt tgccggagct ggtcgcgctg ggctggacgg taaccgagtt
     4441 cgcggcgggc aagtacgaca tcacccggcc caaggcggca ggctgacccc ccccactcta
     4501 ttgtaaacaa gacattttta tcttttatat tcaatggctt atttgctcgg gacgcacggc
     4561 gcgctctacg aactgccgat aaacagagga ttaaaattga caattctagg gcgcgtatag
     4621 cttgccggaa gtcgaaagcc acgttgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     4681 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     4741 tgttatgagc catattcaac gggaaacgtc ttgctcgagg ccgcgattaa attccaacat
     4801 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     4861 aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     4921 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     4981 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     5041 tgcgatcccc gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     5101 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     5161 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     5221 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     5281 gaaagaaatg cataagcttt tgccattctc accggattca gtcgtcactc atggtgattt
     5341 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     5401 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     5461 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     5521 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     5581 gtaacactgg cagagcatta cgctgacttg acgggacggc ggctttgttg aataaatcga
     5641 acttttgctg agttgaagga tcagatcacg cgccacatga ggcatgttcc gcgtccttgc
     5701 aatactgtgt ttacatacag tctatcgctt agcggaaagt tcttttaccc tcagccgaaa
     5761 tgcctgccgt tgctagacat tgccagccag tgcccgtcac tcccgtacta actgtcacga
     5821 acccctgcaa taactgtcac gcccccctgc aataactgtc acgaacccct gcaataactg
     5881 tcacgccccc aaacctgcaa acccagcagg ggcgggggct ggcggggtgt tggaaaaatc
     5941 catccatgat tatctaagaa taatccacta ggcgcggtta tcagcgccct tgtggggcgc
     6001 tgctgccctt gcccaatatg cccggccaga ggccggatag ctggtctatt cgctgcgcta
     6061 ggctacacac cgccccaccg ctgcgcggca gggggaaagg cgggcaaagc ccgctaaacc
     6121 ccacaccaaa ccccgcagaa atacgctgga gcgcttttag ccgctttagc ggcctttccc
     6181 cctacccgaa gggtgggggc gcgtgtgcag ccccgcaggg cctgtctcgg tcgatcattc
     6241 agcccggctc atccttctgg cgtggcggca gaccgaacaa ggcgcggtcg tggtcgcgtt
     6301 caaggtacgc atccattgcc gccatgagcc gatcctccgg ccactcgctg ctgttcacct
     6361 tggccaaaat catggccccc accagcacct tgcgccttgt ttcgttcttg cgctcttgct
     6421 gctgttccct tgcccgcacc cgctgaattt cggcattgat tcgcgctcgt tgttcttcga
     6481 gcttggccag ccgatccgcc gccttgttgc tccccttaac catcttgaca ccccattgtt
     6541 aatgtgctgt ctc
//
