LOCUS       sequence-330131-        2969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  backbone vector for clone construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2969)
  AUTHORS   Wang L, Xiao Y, Wei X, Pan J, Duanmu D
  TITLE     Highly Efficient CRISPR-Mediated Base Editing in Sinorhizobium 
            meliloti.
  JOURNAL   Front Microbiol. 2021 Jun 18;12:686008. doi: 
            10.3389/fmicb.2021.686008. eCollection 2021.
  PUBMED    34220774
REFERENCE   2  (bases 1 to 2969)
  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: "Front 
            Microbiol."; date: "2021-06-18"; volume: "12:686008. doi"; pages: " 
            10.3389/fmicb.2021.686008. eCollection 2021"
COMMENT     Sequence Label: pKm
FEATURES             Location/Qualifiers
     source          1..2969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      30..124
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(223..242)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             234..1049
                     /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"
     oriT            1200..1308
                     /note="incP origin of transfer"
     CDS             complement(1319..1981)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      1982..2753
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     terminator      complement(2858..2944)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tctagaggat ccccgggtac ccgctcttgg actcctgttg atagatccag taatgacctc
       61 agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga
      121 gaatccaggc aattgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
      181 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
      241 atattcagcg tgaaacgagc tgtagccgtc cgcgtctgaa cagcaacatg gatgcggatc
      301 tgtatggcta taaatgggcg cgtgataacg tgggtcagag cggcgcgacc atttatcgtc
      361 tgtatggcaa accggatgcg ccggaactgt ttctgaaaca tggcaaaggc agcgtggcga
      421 acgatgtgac cgatgaaatg gtgcgtctga actggctgac cgaatttatg ccgctgccga
      481 ccattaaaca ttttattcgc accccggatg atgcgtggct gctgaccacc gcgattccgg
      541 gcaaaaccgc gtttcaggtg ctggaagaat atccggatag cggcgaaaac attgtggatg
      601 cgctggccgt gtttctgcgt cgtctgcata gcattccggt gtgcaactgc ccgtttaaca
      661 gcgatcgtgt gtttcgtctg gcccaggcgc agagccgtat gaacaacggc ctggtggatg
      721 cgagcgattt tgatgatgaa cgtaacggct ggccggtgga acaggtgtgg aaagaaatgc
      781 ataaactgct gccgtttagc ccggatagcg tggtgaccca cggcgatttt agcctggata
      841 acctgatttt cgatgaaggc aaactgattg gctgcattga tgtgggccgt gtgggcattg
      901 cggatcgtta tcaggatctg gccattctgt ggaactgcct gggcgaattt agcccgagcc
      961 tgcaaaaacg tctgtttcag aaatatggca ttgataatcc ggatatgaac aaactgcaat
     1021 ttcatctgat gctggatgaa tttttctaat aattaattga actacttttc cgctgcataa
     1081 ccctgcttcg gggtcattat agcgattttt tcggtatatc catccttttt cgcacgatat
     1141 acaggatttt gccaaagggt tcgtgtagac tttccttggt gtatccaacg gcgtcagccg
     1201 ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt ccttcttcac tgtcccttat
     1261 tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag gctggccgta ggccggccct
     1321 accggcgcgg cagcgttacc cgtgtcggcg gctccaacgg ctcgccatcg tccagaaaac
     1381 acggctcatc gggcatcggc aggcgctgct gcccgcgccg ttcccattcc tccgtttcgg
     1441 tcaaggctgg caggtctggt tccatgcccg gaatgccggg ctggctgggc ggctcctcgc
     1501 cggggccggt cggtagttgc tgctcgcccg gatacagggt cgggatgcgg cgcaggtcgc
     1561 catgccccaa cagcgattcg tcctggtcgt cgtgatcaac caccacggcg gcactgaaca
     1621 ccgacaggcg caactggtcg cggggctggc cccacgccac gcggtcattg accacgtagg
     1681 ccgacacggt gccggggccg ttgagcttca cgacggagat ccagcgctcg gccaccaagt
     1741 ccttgactgc gtattggacc gtccgcaaag aacgtccgat gagcttggaa agagtctttt
     1801 ggctgaccac cacggcgttc tggtggccca tctgcgcgac gaggtgatgc agcagcattg
     1861 ccgccgtggg tttcctcgca ataagcccgg cccacgcctc atgcgctttg cgttccgttt
     1921 ggacccagtg accgggcttg ttcttggctt gaatgccgat ttctctggac tgcgtggcca
     1981 tgcttatctc catgcggtag gggtgccgca cggttgcggc accatgcgca atcagctgca
     2041 acttttcggc agcgcgacaa caattatgcg ttgcgtaaaa gtggcagtca attacagatt
     2101 ttctttaacc tacgcaatga gctattgcgg ggggtgccgc aatgagctgt tgcgtacccc
     2161 ccttttttaa gttgttgatt tttaagtctt tcgcatttcg ccctatatct agttctttgg
     2221 tgcccaaaga agggcacccc tgcggggttc ccccacgcct tcggcgcggc tccccctccg
     2281 gcaaaaagtg gcccctccgg ggcttgttga tcgactgcgc ggccttcggc cttgcccaag
     2341 gtggcgctgc ccccttggaa cccccgcact cgccgccgtg aggctcgggg ggcaggcggg
     2401 cgggcttcgc ccttcgactg cccccactcg cataggcttg ggtcgttcca ggcgcgtcaa
     2461 ggccaagccg ctgcgcggtc gctgcgcgag ccttgacccg ccttcctctt ggtgtccaac
     2521 cggcaagcga agcgcgcagg ccgcaggccg gaggcttttc cccagagaaa attaaaaaaa
     2581 ttgatggggc aaggccgcag gccgcgcagt tggagccggt gggtatgtgg tcgaaggctg
     2641 ggtagccggt gggcaatccc tgtggtcaag ctcgtgggca ggcgcagcct gtccatcagc
     2701 ttgtccagca gggttgtcca cgggccgagc gaagcgagcc agccggtggc cgctcgcggc
     2761 catcgtccac atatccacgg gctggcaagg gagcgcagcg accgcgcagg gcgaagcccg
     2821 gagagcaagc ccgtaggggg ttactctagg gcggcggatt tgtcctactc aggagagcgt
     2881 tcaccgacaa acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta
     2941 tttgatgcct gccgaattcg gatccggag
//
