LOCUS       sequence-138908-        4885 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid containing streptomycin resistance gene cassette flanked by 
            multiple restriction sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4885)
  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 4885)
  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: pRGD-SmR
FEATURES             Location/Qualifiers
     source          1..4885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(815..1606)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     primer_bind     complement(2502..2518)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(3066..3654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3146..3165)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3825..4616,4617..4685))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4448..4467
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4686..4790)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4858..4876
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttcgaaca tatggtacca
      421 tcgatgattc actagtgata gctttatgct tgtaaaccgt tttgtgaaaa aatttttaaa
      481 ataaaaaagg ggacctctag ggtccccaat taattagtaa tataatctat taaaggtcat
      541 tcaaaaggtc atccaccgga tcagcttagt aaagccctcg ctagatttta atgcggatgt
      601 tgcgattact tcgccaacta ttgcgataac aagaaaaagc cagcctttca tgatatatct
      661 cccaatttgt gtagggctta ttatgcacgc ttaaaaataa taaaagcaga cttgacctga
      721 tagtttggct gtgagcaatt atgtgcttag tgcatctaac gcttgagtta agccgcgccg
      781 cgaagcggcg tcggcttgaa cgaattgtta gacattattt gccgactacc ttggtgatct
      841 cgcctttcac gtagtggaca aattcttcca actgatctgc gcgcgaggcc aagcgatctt
      901 cttcttgtcc aagataagcc tgtctagctt caagtatgac gggctgatac tgggccggca
      961 ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc gattttgccg gttactgcgc
     1021 tgtaccaaat gcgggacaac gtaagcacta catttcgctc atcgccagcc cagtcgggcg
     1081 gcgagttcca tagcgttaag gtttcattta gcgcctcaaa tagatcctgt tcaggaaccg
     1141 gatcaaagag ttcctccgcc gctggaccta ccaaggcaac gctatgttct cttgcttttg
     1201 tcagcaagat agccagatca atgtcgatcg tggctggctc gaagatacct gcaagaatgt
     1261 cattgcgctg ccattctcca aattgcagtt cgcgcttagc tggataacgc cacggaatga
     1321 tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag aatctcgctc tctccagggg
     1381 aagccgaagt ttccaaaagg tcgttgatca aagctcgccg cgttgtttca tcaagcctta
     1441 cggtcaccgt aaccagcaaa tcaatatcac tgtgtggctt caggccgcca tccactgcgg
     1501 agccgtacaa atgtacggcc agcaacgtcg gttcgagatg gcgctcgatg acgccaacta
     1561 cctctgatag ttgagtcgat acttcggcga tcaccgcttc cctcatgatg tttaactttg
     1621 ttttagggcg actgccctgc tgcgtaacat cgttgctgct ccataacatc aaacatcgac
     1681 ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtacc ccaaaaaaac
     1741 agtcataaca agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg
     1801 ttctggacca gttgcgtgag cgcatacgct acttgcatta cagcttacga accgaacagg
     1861 cttatgtcca ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct
     1921 tgggcagcag cgaagtcgag gcatttctgt cctggctggc gaacgagcgc aaggtttcgg
     1981 tctccacgca tcgtcaggca ttggcggcct tgctgttctt ctacggcaag gtgctgtgca
     2041 cggatctgcc ctggcttcag gagatcggaa gacctcggcc gtcgcggcgc ttgccggtgg
     2101 tgctgacccc ggatgaagtg gttcgcatcc tcggttttct ggaaggcgag catcgtttgt
     2161 tcgcccagct tctgtatgga acgggcatgc ggatcagtga gggtttgcaa ctgcgggtca
     2221 aggatctgga tttcgatcac ggcacgatca tcgtgcggga gggcaagggc tccaaggatc
     2281 gggccttgat gttacccgag agcttggcac ccagcctgcg cgagcagggg aattgatccg
     2341 gtggatgacc ttttgaatga cctttaatag attatattac taattaattg gggaccctag
     2401 aggtcccctt ttttatttta aaaatttttt cacaaaacgg tttacaagca taaagctatc
     2461 ttcgaacgat actagtcata tgaattcggt accagcttgg cactggccgt cgttttacaa
     2521 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     2581 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     2641 agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     2701 tcacaccgca taactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     2761 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     2821 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     2881 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     2941 cgccagccga actgttcgcc aggctcaagg cgcggatgcc cgacggcgag gatctcgtcg
     3001 tgacccatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3061 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3121 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3181 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3241 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3301 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3361 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3421 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3481 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     3541 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3601 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3661 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3721 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3781 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3841 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3901 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3961 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4021 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4081 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4141 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4201 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4261 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4321 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4381 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4441 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4501 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4561 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4621 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4681 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4741 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4801 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     4861 aggcgtatca cgaggccctt tcgtc
//
