LOCUS       sequence-138668-        5348 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mCherry-encoding vector for marker replacement mutagenesis in 
            Bradyrhizobium diazoefficiens and other Alphaproteobacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5348)
  AUTHORS   Ledermann R, Strebel S, Kampik C, Fischer HM
  TITLE     Versatile vectors for efficient mutagenesis of Bradyrhizobium 
            diazoefficiens and other Alphaproteobacteria.
  JOURNAL   Appl Environ Microbiol. 2016 Feb 26. pii: AEM.04085-15.
  PUBMED    26921431
REFERENCE   2  (bases 1 to 5348)
  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. 2016 Feb 26. pii: AEM.04085-15."
COMMENT     Sequence Label: pREDSIX
FEATURES             Location/Qualifiers
     source          1..5348
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(73..90)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(244..832)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(324..343)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1757..2128)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            2161..2270
                     /note="incP origin of transfer"
     primer_bind     complement(3216..3235)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(join(3372..4163,4164..4232))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3995..4014
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4233..4337)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4620..5327
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4765..4783)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5033..5052
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 ctgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
       61 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
      121 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      181 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      241 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      301 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      361 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      421 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      481 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      541 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      601 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      661 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      721 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      781 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      841 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      901 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      961 taaatcaatc taaagtatat atgagtaaac ttggtctgac aggtttaaac tctagacaat
     1021 tgcctaggat gcatgctagc cttaagattt aaatgttaat taatccggtt tattgactac
     1081 cggaagcagt gtgaccgtgt gcttctcaaa tgcctgaggc cagtttgctc aggctctccc
     1141 cgtggaggta ataattgacg atatgatcca gccgaccagg ctttccacgc ccgcgtgccg
     1201 ctccatgtcg ttcgcgcggt tctcggaaac gcgctgccgc gtttcgtgat tgtcacgctc
     1261 aagcccgtag tcccgttcga gcgtcgcgca gaggtcagcg agggcgcggt aggcccgata
     1321 cggctcatgg atggtgtttc gggtcgggtg aatcttgttg atggcgatat ggatgtgcag
     1381 gttgtcggtg tcgtgatgca cggcactgac gcgctgatgc tcggcgaagc caagcccagc
     1441 gcagatgcgg tcctcaatcg cgcgcaacgt ctccgcgtcg ggcttctctc ccgcgcggaa
     1501 gctaaccagc aggtgatagg tcttgtcggc ctcggaacgg gtgttgccgt gctgggtcgc
     1561 catcacctcg gccatgacag cgggcagggt gtttgcctcg cagttcgtga cgcgcacgtg
     1621 acccaggcgc tcggtcttgc cttgctcgtc ggtgatgtac ttcaccagct ccgcgaagtc
     1681 gctcttcttg atggagcgca tggggacgtg cttggcaatc acgcgcaccc cccggccgtt
     1741 ttagcggcta aaaaagtcat ggctctgccc tcgggcggac cacgcccatc atgaccttgc
     1801 caagctcgtc ctgcttctct tcgatcttcg ccagcagggc gaggatcgtg gcatcaccga
     1861 accgcgccgt gcgcgggtcg tcggtgagcc agagtttcag caggccgccc aggcggccca
     1921 ggtcgccatt gatgcgggcc agctcgcgga cgtgctcata gtccacgacg cccgtgattt
     1981 tgtagccctg gccgacggcc agcaggtagg ccgacaggct catgccggcc gccgccgcct
     2041 tttcctcaat cgctcttcgt tcgtctggaa ggcagtacac cttgataggt gggctgccct
     2101 tcctggttgg cttggtttca tcagccatcc gcttgccctc atctgttacg ccggcggtag
     2161 ccggccagcc tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa taagggacag
     2221 tgaagaagga acacccgctc gcgggtgggc ctacttcacc tatcctgccc ggctgacgcc
     2281 gttggataca ccaaggaaag tctacacgaa ccctttggca aaatcctgta tatcgtgcga
     2341 aaaaggatgg atataccgaa aaaatcgcta taatgacccc gaagcagggt tatgcagcgg
     2401 aaaagcgctg cttccctgct gttttgtgga atatctaccg actggaaaca ggcaaatgca
     2461 ggaaattact gaactgaggg gacaggcgag agacgatgcc aaagagctac accgacgagc
     2521 tggccgagtg ggttgaatcc cgcgcggcca agaagcgccg gcgtgatgag gctgcggttg
     2581 cgttcctggc ggtgagggcg gatgtcgagg cggcgttagc gtccggctat gcgctcgtca
     2641 ccatttggga gcacatgcgg gaaacgggga aggtcaagtt ctcctacgag acgttccgct
     2701 cgcacgccag gcggcacatc aaggccaagc ccgccgatgt gcccgcaccg caggccaagg
     2761 ctgcggaacc cgcgccggca cccaagacgc cggagccacg gcggccgaag caggggggca
     2821 aggctgaaaa gccggccccc gctgcggccc cgaccggctt caccttcaac ccaacaccgg
     2881 acaaaaagga tctactgtaa tggcgaaaat tcacatggtt ttgcagggca agggcggggt
     2941 cggcaagtcg gccatcgccg cgatcattgc gcagtacaag atggacaagg ggcagacacc
     3001 cttgtgcatc gacaccgacc cggtgaacgc gacgttcgag ggctacaagg ccctgaacgt
     3061 ccgccggctg aacatcatgg ccggcgacga aattaactcg cgcaacttcg acaccctggt
     3121 cgagctgatt gcgccgacca aggatgacgt ggtgatcctc tacgccggac gcatcgtggc
     3181 cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg
     3241 ggaagatcct tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg
     3301 caacgcgggc atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca
     3361 gcctcgcgtc gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3421 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3481 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3541 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3601 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3661 agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt
     3721 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3781 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3841 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     3901 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     3961 cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact
     4021 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4081 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4141 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4201 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4261 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4321 caaatagggg ttccgcgcac atttccaatt cgcccttagt gaagacaaag gtaagcttgc
     4381 tcgatctcgg cgagcgccgc ggcggcgccc cggaggcgca gcggctctac gagcaggtcg
     4441 gcgaggccgc gtcctgaccc ctcattcggc gcgatccgcc ctgccggcaa acctgccgct
     4501 tgatcatcgt cgcacaagcc tctaatcggt ccgcgccttt acgcccggct tatggccggt
     4561 cccgcggtag gattgcacga agcttagatc ttgactagaa ataagaagga gatataatta
     4621 tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg
     4681 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     4741 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
     4801 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc
     4861 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
     4921 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
     4981 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
     5041 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg
     5101 ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg
     5161 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca
     5221 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
     5281 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaaggaa ttgaagctta
     5341 gatcttga
//
