LOCUS       sequence-472826-        3475 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for disruption of the phaC1 locus of Cupriavidus necator via
            homologous recombination.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3475)
  AUTHORS   Arhar S, Pirchner J, Stolterfoht-Stock H, Reicher K, Kourist R, 
            Emmerstorfer-Augustin A
  TITLE     CnRed: Efficient, Marker-free Genome Engineering of Cupriavidus 
            necator H16 by Adapted Lambda Red Recombineering.
  JOURNAL   ACS Synth Biol. 2025 Feb 24. doi: 10.1021/acssynbio.4c00757.
  PUBMED    39989320
REFERENCE   2  (bases 1 to 3475)
  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: "ACS Synth 
            Biol. 2025 Feb 24. doi: 10.1021/acssynbio.4c00757."
COMMENT     Sequence Label: pHRep-HKH1
FEATURES             Location/Qualifiers
     source          1..3475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(880..899)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             891..1706
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             891..1706
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(964..983)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(2756..3344)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2836..2855)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggtcccgcc agtggcaggg cactgaaggc aacggccacg cggccgcgtc cggcattccg
       61 ggcctggatg cgctggcagg cgtcaagatc gcgccggcgc agctgggtga tatccagcag
      121 cgctacatga aggacttctc agcgctgtgg caggccatgg ccgagggcaa ggccgaggcc
      181 accggtccgc tgcacgaccg gcgcttcgcc ggcgacgcat ggcgcaccaa cctcccatat
      241 cgcttcgctg ccgcgttcta cctgctcaat gcgcgcgcct tgaccgagct ggccgatgcc
      301 gtcgaggccg atgccaagac ccgccagcgc atccgcttcg cgatctcgca atgggtcgat
      361 gcgatgtcgc ccgccaactt ccttgccacc aatcccgagg cgcagcgcct gctgatcgag
      421 tcgggcggcg aatcgctgcg tgccggcgtg cgcaacatga tggaagacct gacacgcggc
      481 aagatctcgc agaccgacga gagcgcgttt gaggtcggcc gcaatgtcgc ggtgaccgaa
      541 ggcgccgtgg tcttcgagaa cgagtacttc cagctgttgc agtacaagcc gctgaccgac
      601 aaggtgcacg cgcgcccgct gctgatggtg ccgccgtgca tcaacaagta ctacatcctg
      661 gacctgcagc cggagagctc gctggtgcgc catgtggtgg agcagggaca tacggtgttt
      721 ctggtgtcgt ggcgcaatcc ggacgccagc atggccggca gcacctggga cgactgaatt
      781 ctttgcgttt ctacaaactc ttttgtttat ttttctaaat acattcaata tgtatccgct
      841 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
      901 ttcaacggga aacgtcttgc tctaggccgc gattaaattc caacatggat gctgatttat
      961 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt
     1021 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     1081 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     1141 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggga
     1201 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     1261 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     1321 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     1381 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     1441 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     1501 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     1561 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     1621 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     1681 atttgatgct cgatgagttt ttctaagaat taattcatga ccaaaatccc ttaacgtgag
     1741 ttttcgttcc actgagcgtc agacaggcct cctaggacgg atggccttgg atccacatcg
     1801 agcacgcggc catccgcgcc atcgaagtcg cgcgcgacat cagcggccag gacaagatca
     1861 acgtgctcgg cttctgcgtg ggcggcacca ttgtctcgac cgcgctggcg gtgctggccg
     1921 cgcgcggcga gcacccggcc gccagcgtca cgctgctgac cacgctgctg gactttgccg
     1981 acacgggcat cctcgacgtc tttgtcgacg agggccatgt gcagttgcgc gaggccacgc
     2041 tgggcggcgg cgccggcgcg ccgtgcgcgc tgctgcgcgg ccttgagctg gccaatacct
     2101 tctcgttctt gcgcccgaac gacctggtgt ggaactacgt ggtcgacaac tacctgaagg
     2161 gcaacacgcc ggtgccgttc gacctgctgt tctggaacgg cgacgccacc aacctgccgg
     2221 ggccgtggta ctgctggtac ctgcgccaca cctacctgca gaacgagctc aaggtaccgg
     2281 gcaagctgac cgtgtgcggc gtgccggtgg acctggccag catcgacgtg ccgacctata
     2341 tctacggctc gcgcgaagac catatcgtgc cgtggaccgc ggcctatgcc tcgaccgcgc
     2401 tgctggcgaa caagctgcgc ttcgtgctgg gtgcgtcggg ccatatcgcc ggtgtgatca
     2461 acccgccggc caagaacaag cgcagccact ggactaacga tgcgctgccg gagtcgccgc
     2521 agcaatggct ggccggcgcc atcgagcatc acggcagctg gtggccggac tggaccgcat
     2581 ggctggccgg gcaggccggc gcgaaacgcg ccgcgcccgc caactatggc aatgcgcgct
     2641 atcgcgcaat cgaacccgcg cctgggcgat acgtcaaagc caaggcatga ctcgagatgt
     2701 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2761 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2821 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2881 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2941 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3001 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3061 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3121 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3181 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3241 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3301 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3361 tttctacggg tctagaatgg cgaccggcaa aggcgcggca gcttccacgc aggaaggcaa
     3421 gtcccaacca ttcaaggtca cgccggggcc attcgatcca gccacatggc tggaa
//
