LOCUS       sequence-358835-        3068 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Spacer sequence in 2.2b format to allow for level 3 cloning with 
            D2.2a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3068)
  AUTHORS   Goosens VJ, Walker KT, Aragon SM, Singh A, Senthivel VR, Dekker L, 
            Caro-Astorga J, Buat MLA, Song W, Lee KY, Ellis T
  TITLE     Komagataeibacter Tool Kit (KTK): A Modular Cloning System for 
            Multigene Constructs and Programmed Protein Secretion from Cellulose
            Producing Bacteria.
  JOURNAL   ACS Synth Biol. 2021 Dec 17;10(12):3422-3434. doi: 
            10.1021/acssynbio.1c00358. Epub 2021 Nov 12.
  PUBMED    34767345
REFERENCE   2  (bases 1 to 3068)
  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.1c00358"; journalName: "ACS Synth Biol"; date: 
            "2021-12-17- 17"; volume: "10"; issue: "12"; pages: "3422-3434"
COMMENT     Sequence Label: KTK_324
FEATURES             Location/Qualifiers
     source          1..3068
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(466..552)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      607..701
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(733..1524)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     oriT            1866..1974
                     /note="incP origin of transfer"
     CDS             complement(1985..2647)
                     /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(2648..3068,1..351)
                     /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 ggcttcgccc ttcgactgcc cccactcgca taggcttggg tcgttccagg cgcgtcaagg
       61 ccaagccgct gcgcggtcgc tgcgcgagcc ttgacccgcc ttccacttgg tgtccaaccg
      121 gcaagcgaag cgcgcaggcc gcaggccgga ggcttttccc cagagaaaat taaaaaaatt
      181 gatggggcaa ggccgcaggc cgcgcagttg gagccggtgg gtatgtggtc gaaggctggg
      241 tagccggtgg gcaatccctg tggtcaagct cgtgggcagg cgcagcctgt ccatcagctt
      301 gtccagcagg gttgtccacg ggccgagcga agcgagccag ccggtggccg ctcgcggcca
      361 tcgtccacat atccacgggc tggcaaggga gcgcagcgac cgcgcagggc gaagcccgga
      421 gagcaagccc gtaggggggg cgcgcccagc tgtctagggc ggcggatttg tcctactcag
      481 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtctttcg actgagcctt
      541 tcgttttatt tgatgccttt aattaaggtc tcgtatgcga cccgaaacgt gagaccacta
      601 gtcttggact cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca
      661 gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaggggtc cccaataatt
      721 acgatttttg tcttatttgc cgactacctt ggtgatctcg cctttcacgt agtggacaaa
      781 ttcttccaac tgatctgcgc gcgaggccaa gcgatcttct tcttgtccaa gataagcctg
      841 tctagcttca agtatgacgg gctgatactg ggccggcagg cgctccattg cccagtcggc
      901 agcgacatcc ttcggcgcga ttttgccggt tactgcgctg taccaaatgc gggacaacgt
      961 aagcactaca tttcgctcat cgccagccca gtcgggcggc gagttccata gcgttaaggt
     1021 ttcatttagc gcctcaaata gatcctgttc aggaaccgga tcaaagagtt cctccgccgc
     1081 tggacctacc aaggcaacgc tatgttctct tgcttttgtc agcaagatag ccagatcaat
     1141 gtcgatcgtg gctggctcga agatacctgc aagaatgtca ttgcgctgcc attctccaaa
     1201 ttgcagttcg cgcttagctg gataacgcca cggaatgatg tcgtcgtgca caacaatggt
     1261 gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt ccaaaaggtc
     1321 gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa ccagcaaatc
     1381 aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat gtacggccag
     1441 caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt gagtcgatac
     1501 ttcggcgatc accgcttccc tcatgatgtt taactttgtt ttagggcgac tgccctgctg
     1561 cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc acggcgtaac gcgcttgctg
     1621 cttggatgcc cgaggcatag actgtacccc aaaaaaacag tcataacaag ccatgaaaac
     1681 cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt catttttgtc cttttccgct
     1741 gcataaccct gcttcggggt cattatagcg attttttcgg tatatccatc ctttttcgca
     1801 cgatatacag gattttgcca aagggttcgt gtagactttc cttggtgtat ccaacggcgt
     1861 cagccgggca ggataggtga agtaggccca cccgcgagcg ggtgttcctt cttcactgtc
     1921 ccttattcgc acctggcggt gctcaacggg aatcctgctc tgcgaggctg gccgtaggcc
     1981 ggccctaccg gcgcggcagc gttacccgtg tcggcggctc caacggctcg ccatcgtcca
     2041 gaaaacacgg ctcatcgggc atcggcaggc gctgctgccc gcgccgttcc cattcctccg
     2101 tttcggtcaa ggctggcagg tctggttcca tgcccggaat gccgggctgg ctgggcggct
     2161 cctcgccggg gccggtcggt agttgctgct cgcccggata cagggtcggg atgcggcgca
     2221 ggtcgccatg ccccaacagc gattcgtcct ggtcgtcgtg atcaaccacc acggcggcac
     2281 tgaacaccga caggcgcaac tggtcgcggg gctggcccca cgccacgcgg tcattgacca
     2341 cgtaggccga cacggtgccg gggccgttga gcttcacgac ggagatccag cgctcggcca
     2401 ccaagtcctt gactgcgtat tggaccgtcc gcaaagaacg tccgatgagc ttggaaagtg
     2461 ttttctggct gaccaccacg gcgttctggt ggcccatctg cgccacgagg tgatgcagca
     2521 gcattgccgc cgtgggtttc ctcgcaataa gcccggccca cgcctcatgc gctttgcgtt
     2581 ccgtttgcac ccagtgaccg ggcttgttct tggcttgaat gccgatttct ctggactgcg
     2641 tggccatgct tatctccatg cggtaggggt gccgcacggt tgcggcacca tgcgcaatca
     2701 gctgcaactt ttcggcagcg cgacaacaat tatgcgttgc gtaaaagtgg cagtcaatta
     2761 cagattttct ttaacctacg caatgagcta ttgcgggggg tgccgcaatg agctgttgcg
     2821 tacccccctt ttttaagttg ttgattttta agtctttcgc atttcgccct atatctagtt
     2881 ctttggtgcc caaagaaggg cacccctgcg gggttccccc acgccttcgg cgcggctccc
     2941 cctccggcaa aaagtggccc ctccggggct tgttgatcga ctgcgcggcc ttcggccttg
     3001 cccaaggtgg cgctgccccc ttggaacccc cgcactcgcc gccgtgaggc tcggggggca
     3061 ggcgggcg
//
