LOCUS       sequence-304718-        4271 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid containing hygromycin B resistance gene cassette flanked by 
            multiple restriction sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4271)
  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 4271)
  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-HmR
FEATURES             Location/Qualifiers
     source          1..4271
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(17..33)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(242..261)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     361..383
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(421..439)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        507..611
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(612..680,681..1472)
                     /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     complement(830..849)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1643..2231
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2132..2151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2779..2795
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2872..3897)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFGSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEVMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDPLYEVANIFFWRPWLACMEQQARYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
ORIGIN
        1 atgttcgaag cttggcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
       61 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
      121 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga
      181 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
      241 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
      301 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
      361 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
      421 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
      481 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
      541 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      601 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
      661 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
      721 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
      781 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
      841 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
      901 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
      961 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1021 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     1081 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     1141 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     1201 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     1261 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     1321 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     1381 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     1441 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     1501 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     1561 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1621 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1681 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1741 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     1801 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1861 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1921 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1981 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2041 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2101 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2161 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     2221 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     2281 ttgctcacat gggtcacgac gagatcctcg ccgtcgggca tccgcgcctt gagcctggcg
     2341 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
     2401 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
     2461 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
     2521 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
     2581 cagttatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     2641 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     2701 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     2761 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctggtaccg aattcatatg
     2821 actagtatcg ttcgaagatc gggtgagcac cgaacccacc atgatttccg tctattcctt
     2881 tgccctcgga cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc
     2941 atcggtccag acggccgcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc
     3001 ggatcggacg attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc
     3061 aaccaagctc tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagccgcgg
     3121 cgatcctgca agctccggat gcctccgctc gaagtagcgc gcctgctgct ccatacaagc
     3181 caaccacggc ctccagaaga agatgttggc gacctcgtat aggggatccc cgaacatcgc
     3241 ctcgctccag tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc
     3301 gaaatccgcg tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc
     3361 atcgagagcc tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata
     3421 cacatgggga tcagcaatcg cgcagatgaa atcacgccat gtagtgtatt gaccgattcc
     3481 ttgcggtccg aatgggccga acccgctcgt ctggctaaga tcggccgcag cgatcgcatc
     3541 cataacctcc gcgaccggtt gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa
     3601 cgtgacaccc tgtgcacggc gggagatgca ataggtcagg ctctcgctaa attccccaat
     3661 gtcaagcact tccggaatcg ggagcgcggc cgatgcaaag tgccgataaa cataacgatc
     3721 tttgtagaaa ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc
     3781 gaagctgaaa gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgcc
     3841 gaacttttcg atcagaaact tctcaacaga cgtcgcggtg agttcaggct ttttcatact
     3901 agtatcctgt ctcttgatca gatcttgatc ccctgcgcca tcagatcctt ggcggcaaga
     3961 aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca
     4021 attccggttc gcttgctgtc cataaaaccg cccagtctag ctatcgccat gtaagcccac
     4081 tgcaagctac ctgctttctc tttgcgcttg cgttttccct tgtccagata gcccagtagc
     4141 tgacattcat ccggggtcag caccgtttct gcggactggc tttctacgtg ttccgcttcc
     4201 tttagcagcc cttgcgccct gagtgcttgc ggcagcgtgc gagctcgaat tccgtatatc
     4261 gatggtacca t
//
