LOCUS       sequence-404757-        1782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning plasmid containing the kanamycin resistance gene insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1782)
  AUTHORS   van Schaik J, Li Z, Cheadle J, Crook N
  TITLE     Engineering the Maize Root Microbiome: A Rapid MoClo Toolkit and 
            Identification of Potential Bacterial Chassis for Studying 
            Plant-Microbe Interactions.
  JOURNAL   ACS Synth Biol. 2023 Oct 20;12(10):3030-3040. doi: 
            10.1021/acssynbio.3c00371. Epub 2023 Sep 15.
  PUBMED    37712562
REFERENCE   2  (bases 1 to 1782)
  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"; doi: 
            "10.1021/acssynbio.3c00371"; journalName: "ACS Synth Biol"; date: 
            "2023-10-20- 20"; volume: "12"; issue: "10"; pages: "3030-3040"
COMMENT     Sequence Label: pMRMTK-clo6
FEATURES             Location/Qualifiers
     source          1..1782
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      329..356
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(380..1195)
                     /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             complement(380..1195)
                     /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     1103..1122
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1187..1206
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     oriT            1476..1585
                     /note="incP origin of transfer"
     rep_origin      join(1700..1782,1..306)
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 ctaagctctc atgtttaacg aactaaaccc tcatggctaa cgtactaagc tctcatggct
       61 aacgtactaa gctctcatgt ttcacgtact aagctctcat gtttgaacaa taaaattaat
      121 ataaatcagc aacttaaata gcctctaagg ttttaagttt tataagaaaa aaaagaatat
      181 ataaggcttt taaagctttt aaggtttaac ggttgtggac aacaagccag ggatgtaacg
      241 cactgagaag cccttagagc ctctcaaagc aattttgagt gacacaggaa cacttaacgg
      301 ctgacatggg ggtctcgctc ctcacactgg ctcaccttcg ggtgggcctt tctgcgttta
      361 tacctaggag tcaaaattat tagaaaaact catcgagcat caaatgaaac tgcaatttat
      421 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
      481 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
      541 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
      601 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagcttatgc atttctttcc
      661 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
      721 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
      781 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
      841 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
      901 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
      961 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
     1021 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
     1081 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
     1141 tgttggaatt taatcgcggc ctcgagcaag acgtttcccg ttgaatatgg ctcataacac
     1201 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat
     1261 cttgtgcaat gtaacatcag agattttgag acacaacgtg gctttagcgc gagaccgtca
     1321 attcgagggg gatcaattcc gttttccgct gcataaccct gcttcggggt cattatagcg
     1381 attttttcgg tatatccatc ctttttcgca cgatatacag gattttgcca aagggttcgt
     1441 gtagactttc cttggtgtat ccaacggcgt cagccgggca ggataggtga agtaggccca
     1501 cccgcgagcg ggtgttcctt cttcactgtc ccttattcgc acctggcggt gctcaacggg
     1561 aatcctgctc tgcgaggctg gccggctacc gccggcgtaa cagatgaggg caagcggatg
     1621 gctgatgaaa ccaagccaac caggaagggc agcccaccta tcacggaatt gatccggcca
     1681 cgatgcgtcc ggcgtagagg atctgaagat cagcagttca acctgttgat agtacgtact
     1741 aagctctcat gtttcacgta ctaagctctc atgtttaacg ta
//
