LOCUS       sequence-337560-        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      complement(564..650)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      705..799
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(898..917)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             909..1724
                     /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            1875..1983
                     /note="incP origin of transfer"
     CDS             complement(1994..2656)
                     /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      join(2657..2969,1..459)
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 ctccggggct tgttgatcga ctgcgcggcc ttcggccttg cccaaggtgg cgctgccccc
       61 ttggaacccc cgcactcgcc gccgtgaggc tcggggggca ggcgggcggg cttcgccctt
      121 cgactgcccc cactcgcata ggcttgggtc gttccaggcg cgtcaaggcc aagccgctgc
      181 gcggtcgctg cgcgagcctt gacccgcctt cctcttggtg tccaaccggc aagcgaagcg
      241 cgcaggccgc aggccggagg cttttcccca gagaaaatta aaaaaattga tggggcaagg
      301 ccgcaggccg cgcagttgga gccggtgggt atgtggtcga aggctgggta gccggtgggc
      361 aatccctgtg gtcaagctcg tgggcaggcg cagcctgtcc atcagcttgt ccagcagggt
      421 tgtccacggg ccgagcgaag cgagccagcc ggtggccgct cgcggccatc gtccacatat
      481 ccacgggctg gcaagggagc gcagcgaccg cgcagggcga agcccggaga gcaagcccgt
      541 agggggttac tctagggcgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa
      601 cagataaaac gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctgccg
      661 aattcggatc cggagtctag aggatccccg ggtacccgct cttggactcc tgttgataga
      721 tccagtaatg acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc
      781 gttttttatt ggtgagaatc caggcaattg tgtctcaaaa tctctgatgt tacattgcac
      841 aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa
      901 ggggtgttat gagccatatt cagcgtgaaa cgagctgtag ccgtccgcgt ctgaacagca
      961 acatggatgc ggatctgtat ggctataaat gggcgcgtga taacgtgggt cagagcggcg
     1021 cgaccattta tcgtctgtat ggcaaaccgg atgcgccgga actgtttctg aaacatggca
     1081 aaggcagcgt ggcgaacgat gtgaccgatg aaatggtgcg tctgaactgg ctgaccgaat
     1141 ttatgccgct gccgaccatt aaacatttta ttcgcacccc ggatgatgcg tggctgctga
     1201 ccaccgcgat tccgggcaaa accgcgtttc aggtgctgga agaatatccg gatagcggcg
     1261 aaaacattgt ggatgcgctg gccgtgtttc tgcgtcgtct gcatagcatt ccggtgtgca
     1321 actgcccgtt taacagcgat cgtgtgtttc gtctggccca ggcgcagagc cgtatgaaca
     1381 acggcctggt ggatgcgagc gattttgatg atgaacgtaa cggctggccg gtggaacagg
     1441 tgtggaaaga aatgcataaa ctgctgccgt ttagcccgga tagcgtggtg acccacggcg
     1501 attttagcct ggataacctg attttcgatg aaggcaaact gattggctgc attgatgtgg
     1561 gccgtgtggg cattgcggat cgttatcagg atctggccat tctgtggaac tgcctgggcg
     1621 aatttagccc gagcctgcaa aaacgtctgt ttcagaaata tggcattgat aatccggata
     1681 tgaacaaact gcaatttcat ctgatgctgg atgaattttt ctaataatta attgaactac
     1741 ttttccgctg cataaccctg cttcggggtc attatagcga ttttttcggt atatccatcc
     1801 tttttcgcac gatatacagg attttgccaa agggttcgtg tagactttcc ttggtgtatc
     1861 caacggcgtc agccgggcag gataggtgaa gtaggcccac ccgcgagcgg gtgttccttc
     1921 ttcactgtcc cttattcgca cctggcggtg ctcaacggga atcctgctct gcgaggctgg
     1981 ccgtaggccg gccctaccgg cgcggcagcg ttacccgtgt cggcggctcc aacggctcgc
     2041 catcgtccag aaaacacggc tcatcgggca tcggcaggcg ctgctgcccg cgccgttccc
     2101 attcctccgt ttcggtcaag gctggcaggt ctggttccat gcccggaatg ccgggctggc
     2161 tgggcggctc ctcgccgggg ccggtcggta gttgctgctc gcccggatac agggtcggga
     2221 tgcggcgcag gtcgccatgc cccaacagcg attcgtcctg gtcgtcgtga tcaaccacca
     2281 cggcggcact gaacaccgac aggcgcaact ggtcgcgggg ctggccccac gccacgcggt
     2341 cattgaccac gtaggccgac acggtgccgg ggccgttgag cttcacgacg gagatccagc
     2401 gctcggccac caagtccttg actgcgtatt ggaccgtccg caaagaacgt ccgatgagct
     2461 tggaaagagt cttttggctg accaccacgg cgttctggtg gcccatctgc gcgacgaggt
     2521 gatgcagcag cattgccgcc gtgggtttcc tcgcaataag cccggcccac gcctcatgcg
     2581 ctttgcgttc cgtttggacc cagtgaccgg gcttgttctt ggcttgaatg ccgatttctc
     2641 tggactgcgt ggccatgctt atctccatgc ggtaggggtg ccgcacggtt gcggcaccat
     2701 gcgcaatcag ctgcaacttt tcggcagcgc gacaacaatt atgcgttgcg taaaagtggc
     2761 agtcaattac agattttctt taacctacgc aatgagctat tgcggggggt gccgcaatga
     2821 gctgttgcgt accccccttt tttaagttgt tgatttttaa gtctttcgca tttcgcccta
     2881 tatctagttc tttggtgccc aaagaagggc acccctgcgg ggttccccca cgccttcggc
     2941 gcggctcccc ctccggcaaa aagtggccc
//
