LOCUS       sequence-138907-        4045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid containing kanamycin resistance gene cassette flanked by 
            multiple restriction sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4045)
  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 4045)
  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-KmR
FEATURES             Location/Qualifiers
     source          1..4045
                     /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(466..1260)
                     /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(577..596)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1187..1206
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(1662..1678)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(2226..2814)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2306..2325)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2985..3776,3777..3845))
                     /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     3608..3627
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3846..3950)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4018..4036
                     /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 gcttgggcga accccagagt cccgctcaga agaactcgtc
      481 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag
      541 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat
      601 gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc
      661 attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc
      721 gtcgggcatc cgcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc
      781 ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat
      841 gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg
      901 cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc
      961 ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag
     1021 cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg
     1081 gagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc
     1141 tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc
     1201 gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat
     1261 gcgaaacgat cctcatcctg tctcttgatc agatcttgat cccctgcgcc atcagatcct
     1321 tggcggcaag aaagccatcc agtttacttt gcagggcttc ccaaccttac cagagggcgc
     1381 cccagctggc aattccggtt cgcttgctgt ccataaaacc gcccagtcta gctatcgcca
     1441 tgtaagccca ctgcaagcta cctgctttct ctttgcgctt gcgttttccc ttgtccagat
     1501 agcccagtag ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctacgt
     1561 gttccgcttc ctttagcagc ccttgcgccc tgagtgcttg cggcagcgtg aagctagatc
     1621 ttcgaacgat actagtcata tgaattcggt accagcttgg cactggccgt cgttttacaa
     1681 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     1741 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     1801 agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     1861 tcacaccgca taactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     1921 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     1981 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     2041 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     2101 cgccagccga actgttcgcc aggctcaagg cgcggatgcc cgacggcgag gatctcgtcg
     2161 tgacccatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2221 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2281 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2341 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2401 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2461 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2521 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2581 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2641 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     2701 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2761 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2821 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2881 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2941 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3001 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3061 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3121 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3181 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3241 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3301 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3361 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3421 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3481 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3541 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3601 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3661 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3721 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3781 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3841 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3901 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3961 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     4021 aggcgtatca cgaggccctt tcgtc
//
