LOCUS       sequence-380008-        6421 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A pBBR-1 based vector with IncP group plasmid compatibility for 
            expression in Methylorubrum extorquens using rhamnose dependent 
            induction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6421)
  AUTHORS   Poschel L, Gehr E, Buchhaupt M
  TITLE     A pBBR1-based vector with IncP group plasmid compatibility for 
            Methylorubrum extorquens.
  JOURNAL   Microbiologyopen. 2022 Oct;11(5):e1325. doi: 10.1002/mbo3.1325.
  PUBMED    36314759
REFERENCE   2  (bases 1 to 6421)
  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: 
            "Microbiologyopen."; date: "2022-10"; volume: "11(5):e1325. doi"; 
            pages: " 10.1002/mbo3.1325"
COMMENT     Sequence Label: pMis1_1B
FEATURES             Location/Qualifiers
     source          1..6421
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1078..1849
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1850..2512
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     promoter        complement(2904..3022)
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     CDS             3168..4004
                     /codon_start=1
                     /gene="E. coli rhaS"
                     /product="positive regulator of the rhaB promoter
                     "
                     /label=rhaS
                     /note="activated by L-rhamnose"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     CDS             4078..4926
                     /codon_start=1
                     /gene="E. coli rhaR"
                     /product="transcriptional activator of rhaR and rhaS"
                     /label=rhaR
                     /note="needed for rhamnose-dependent activation of the rhaB
                     promoter"
                     /translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
                     RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
                     PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
                     YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
                     QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
                     D"
     CDS             complement(5225..6019)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5336..5355)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5946..5965
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tcccgattca acggccccag ggcgtccaga acgggcttca ggcgctcccg aaggtctcgg
       61 gccgtctctt gggcttgatc ggccttcttg cgcatctcac gcgctcctgc ggcggcctgt
      121 agggcaggct catacccctg ccgaaccgct tttgtcagcc ggtcggccac ggcttccggc
      181 gtctcaacgc gctttgagat tcccagcttt tcggccaatc cctgcggtgc ataggcgcgt
      241 ggctcgaccg cttgcgggct gatggtgacg tggcccactg gtggccgctc cagggcctcg
      301 tagaacgcct gaatgcgcgt gtgacgtgcc ttgctgccct cgatgccccg ttgcagccct
      361 agatcggcca cagcggccgc aaacgtggtc tggtcgcggg tcatctgcgc tttgttgccg
      421 atgaactcct tggccgacag cctgccgtcc tgcgtcagcg gcaccacgaa cgcggtcatg
      481 tgcgggctgg tttcgtcacg gtggatgctg gccgtcacga tgcgatccgc cccgtacttg
      541 tccgccagcc acttgtgcgc cttctcgaag aacgccgcct gctgttcttg gctggccgac
      601 ttccaccatt ccgggctggc cgtcatgacg tactcgaccg ccaacacagc gtccttgcgc
      661 cgcttctctg gcagcaactc gcgcagtcgg cccatcgctt catcggtgct gctggccgcc
      721 cagtgctcgt tctctggcgt cctgctggcg tcagcgttgg gcgtctcgcg ctcgcggtag
      781 gcgtgcttga gactggccgc cacgttgccc attttcgcca gcttcttgca tcgcatgatc
      841 gcgtatgccg ccatgcctgc ccctcccttt tggtgtccaa ccggctcgac gggggcagcg
      901 caaggcggtg cctccggcgg gccactcaat gcttgagtat actcactaga ctttgcttcg
      961 caaagtcgtg accgcctacg gcggctgcgg cgccctacgg gcttgctctc cgggcttcgc
     1021 cctgcgcggt cgctgcgctc ccttgccagc ccgtggatat gtggacgatg gccgcgagcg
     1081 gccaccggct ggctcgcttc gctcggcccg tggacaaccc tgctggacaa gctgatggac
     1141 aggctgcgcc tgcccacgag cttgaccaca gggattgccc accggctacc cagccttcga
     1201 ccacataccc accggctcca actgcgcggc ctgcggcctt gccccatcaa tttttttaat
     1261 tttctctggg gaaaagcctc cggcctgcgg cctgcgcgct tcgcttgccg gttggacacc
     1321 aagtggaagg cgggtcaagg ctcgcgcagc gaccgcgcag cggcttggcc ttgacgcgcc
     1381 tggaacgacc caagcctatg cgagtggggg cagtcgaagg gcgaagcccg cccgcctgcc
     1441 ccccgagcct cacggcggcg agtgcggggg ttccaagggg gcagcgccac cttgggcaag
     1501 gccgaaggcc gcgcagtcga tcaacaagcc ccggaggggc cactttttgc cggaggggga
     1561 gccgcgccga aggcgtgggg gaaccccgca ggggtgccct tctttgggca ccaaagaact
     1621 agatataggg cgaaatgcga aagacttaaa aatcaacaac ttaaaaaagg ggggtacgca
     1681 acagctcatt gcggcacccc ccgcaatagc tcattgcgta ggttaaagaa aatctgtaat
     1741 tgactgccac ttttacgcaa cgcataattg ttgtcgcgct gccgaaaagt tgcagctgat
     1801 tgcgcatggt gccgcaaccg tgcggcaccc ctaccgcatg gagataagca tggccacgca
     1861 gtccagagaa atcggcattc aagccaagaa caagcccggt cactgggtgc aaacggaacg
     1921 caaagcgcat gaggcgtggg ccgggcttat tgcgaggaaa cccacggcgg caatgctgct
     1981 gcatcacctc gtggcgcaga tgggccacca gaacgccgtg gtggtcagcc agaagacact
     2041 ttccaagctc atcggacgtt ctttgcggac ggtccaatac gcagtcaagg acttggtggc
     2101 cgagcgctgg atctccgtcg tgaagctcaa cggccccggc accgtgtcgg cctacgtggt
     2161 caatgaccgc gtggcgtggg gccagccccg cgaccagttg cgcctgtcgg tgttcagtgc
     2221 cgccgtggtg gttgatcacg acgaccagga cgaatcgctg ttggggcatg gcgacctgcg
     2281 ccgcatcccg accctgtatc cgggcgagca gcaactaccg accggccccg gcgaggagcc
     2341 gcccagccag cccggcattc cgggcatgga accagacctg ccagccttga ccgaaacgga
     2401 ggaatgggaa cggcgcgggc agcagcgcct gccgatgccc gatgagccgt gttttctgga
     2461 cgatggcgag ccgttggagc cgccgacacg ggtcacgctg ccgcgccggt agcacttggg
     2521 ttgcgcagca acccgtaagt gcgctgttcc agactatcgg ctgtagccgc ctcgccgccc
     2581 tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct
     2641 gaatggaagc cggcggcacc tcgctaacgg attcaccgtt tttatcaggc tctgggaggc
     2701 agaataaatg atcatatcgt caattattac ctccacgggg agagcctgag caaactggcc
     2761 tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaagctt
     2821 ggatcctgta cactcgagga gctcgtcgac ggtaccgttt aaactccttc ttatacttaa
     2881 ctaatatact tctagaggtt cattacgacc agtctaaaaa gcgcctgaat tcgcgacctt
     2941 ctcgttactg acaggaaaat gggccattgg caaccaggga aagatgaacg tgatgatgtt
     3001 cacaatttgc tgaattgtgg tgatgtgatg ctcaccgcat ttcctgaaaa ttcacgctgt
     3061 atcttgaaaa atcgacgttt tttacgtggt tttccgtcga aaatttaagg taagaacctg
     3121 acctcgtgat tactatttcg ccgtgttgac gacatcagga ggccagtatg accgtattac
     3181 atagtgtgga tttttttccg tctggtaacg cgtccgtggc gatagaaccc cggctcccgc
     3241 aggcggattt tcctgaacat catcatgatt ttcatgaaat tgtgattgtc gaacatggca
     3301 cgggtattca tgtgtttaat gggcagccct ataccatcac cggtggcacg gtctgtttcg
     3361 tacgcgatca tgatcggcat ctgtatgaac ataccgataa tctgtgtctg accaatgtgc
     3421 tgtatcgctc gccggatcga tttcagtttc tcgccgggct gaatcagttg ctgccacaag
     3481 agctggatgg gcagtatccg tctcactggc gcgttaacca cagcgtattg cagcaggtgc
     3541 gacagctggt tgcacagatg gaacagcagg aaggggaaaa tgatttaccc tcgaccgcca
     3601 gtcgcgagat cttgtttatg caattactgc tcttgctgcg taaaagcagt ttgcaggaga
     3661 acctggaaaa cagcgcatca cgtctcaact tgcttctggc ctggctggag gaccattttg
     3721 ccgatgaggt gaattgggat gccgtggcgg atcaattttc tctttcactg cgtacgctac
     3781 atcggcagct taagcagcaa acgggactga cgcctcagcg atacctgaac cgcctgcgac
     3841 tgatgaaagc ccgacatctg ctacgccaca gcgaggccag cgttactgac atcgcctatc
     3901 gctgtggatt cagcgacagt aaccactttt cgacgctttt tcgccgagag tttaactggt
     3961 caccgcgtga tattcgccag ggacgggatg gctttctgca ataacgcgaa tcttctcaac
     4021 gtatttgtac gccatattgc gaataatcaa cttcgttctc tggccgaggt agccacggtg
     4081 gcgcatcagt taaaacttct caaagatgat ttttttgcca gcgaccagca ggcagtcgct
     4141 gtggctgacc gttatccgca agatgtcttt gctgaacata cacatgattt ttgtgagctg
     4201 gtgattgtct ggcgcggtaa tggcctgcat gtactcaacg atcgccctta tcgcattacc
     4261 cgtggcgatc tcttttacat tcatgctgac gataaacact cctacgcttc cgttaacgat
     4321 ctggttttgc agaatattat ttattgcccg gagcgtctga agctgaatct tgactggcag
     4381 ggggcgattc cgggatttaa cgccagcgca gggcaaccac actggcgctt aggtagcatg
     4441 gggatggcgc aggcgcggca ggttatcggt cagcttgagc atgaaagtag tcagcatgtg
     4501 ccgtttgcta acgaaatggc tgagttgctg ttcgggcagt tggtgatgtt gctgaatcgc
     4561 catcgttaca ccagtgattc gttgccgcca acatccagcg aaacgttgct ggataagctg
     4621 attacccggc tggcggctag cctgaaaagt ccctttgcgc tggataaatt ttgtgatgag
     4681 gcatcgtgca gtgagcgcgt tttgcgtcag caatttcgcc agcagactgg aatgaccatc
     4741 aatcaatatc tgcgacaggt cagagtgtgt catgcgcaat atcttctcca gcatagccgc
     4801 ctgttaatca gtgatatttc gaccgaatgt ggctttgaag atagtaacta tttttcggtg
     4861 gtgtttaccc gggaaaccgg gatgacgccc agccagtggc gtcatctcaa ttcgcagaaa
     4921 gattaatgca taaaaactgt tgtaattcat taagcattct gccgacatgg aagccatcac
     4981 aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat
     5041 atttgcccat gggggtgggc gaagaactcc agcatgagat ccccgcgctg gaggatcatc
     5101 cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc ggcggtggaa
     5161 tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc
     5221 ccgctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg
     5281 cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat
     5341 cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga
     5401 tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg
     5461 tcacgacgag atcctcgccg tcgggcatgc gcgccttgag cctggcgaac agttcggctg
     5521 gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc
     5581 gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat
     5641 caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa
     5701 ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg
     5761 cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata
     5821 gccgcgctgc ctcgtcctgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa
     5881 gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct
     5941 gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca
     6001 atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatca gatcttgatc
     6061 ccctgcgcca tcagatcctt ggcggcaaga aagccatcca gtttactttg cagggcttcc
     6121 caaccttacc agagggcgcc ccagctggca attccggttc gcttgctgtc cataaaaccg
     6181 cccagtctag ctatcgccat gtaagcccac tgcaagctac ctgctttctc tttgcgcttg
     6241 cgttttccct tgtccagata gcccagtagc tgacattcat cccaggtggc acttttcggg
     6301 gaaatgtgcg cgcccgcgtt cctgctggcg ctgggcctgt ttctggcgct ggacttcccg
     6361 ctgttccgtc agcagctttt cgcccacggc cttgatgatc gcggcggcct tggcctgcat
     6421 a
//
