LOCUS       sequence-373326-        2879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Spacer sequence in D1.1 format. Allows for Level 2 cloning with 
            D1.2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2879)
  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 2879)
  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_139
FEATURES             Location/Qualifiers
     source          1..2879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        114..216
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             217..876
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(283..302)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     oriT            1025..1133
                     /note="incP origin of transfer"
     CDS             complement(1144..1806)
                     /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      1807..2578
                     /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(2693..2779)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(2851..2879,1..66)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt
       61 gagaatccag gggtccccaa taattacgat ttaaattggc gaaaatgaga cgttgatcgg
      121 cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt
      181 gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat
      241 accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt
      301 gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta
      361 aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat
      421 gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt
      481 cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa
      541 taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt
      601 gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat
      661 ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc
      721 cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg
      781 attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta
      841 caacagtact gcgatgagtg gcagggcggg gcgtaatttg acttttgtcc ttttccgctg
      901 cataaccctg cttcggggtc attatagcga ttttttcggt atatccatcc tttttcgcac
      961 gatatacagg attttgccaa agggttcgtg tagactttcc ttggtgtatc caacggcgtc
     1021 agccgggcag gataggtgaa gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc
     1081 cttattcgca cctggcggtg ctcaacggga atcctgctct gcgaggctgg ccgtaggccg
     1141 gccctaccgg cgcggcagcg ttacccgtgt cggcggctcc aacggctcgc catcgtccag
     1201 aaaacacggc tcatcgggca tcggcaggcg ctgctgcccg cgccgttccc attcctccgt
     1261 ttcggtcaag gctggcaggt ctggttccat gcccggaatg ccgggctggc tgggcggctc
     1321 ctcgccgggg ccggtcggta gttgctgctc gcccggatac agggtcggga tgcggcgcag
     1381 gtcgccatgc cccaacagcg attcgtcctg gtcgtcgtga tcaaccacca cggcggcact
     1441 gaacaccgac aggcgcaact ggtcgcgggg ctggccccac gccacgcggt cattgaccac
     1501 gtaggccgac acggtgccgg ggccgttgag cttcacgacg gagatccagc gctcggccac
     1561 caagtccttg actgcgtatt ggaccgtccg caaagaacgt ccgatgagct tggaaagtgt
     1621 tttctggctg accaccacgg cgttctggtg gcccatctgc gccacgaggt gatgcagcag
     1681 cattgccgcc gtgggtttcc tcgcaataag cccggcccac gcctcatgcg ctttgcgttc
     1741 cgtttgcacc cagtgaccgg gcttgttctt ggcttgaatg ccgatttctc tggactgcgt
     1801 ggccatgctt atctccatgc ggtaggggtg ccgcacggtt gcggcaccat gcgcaatcag
     1861 ctgcaacttt tcggcagcgc gacaacaatt atgcgttgcg taaaagtggc agtcaattac
     1921 agattttctt taacctacgc aatgagctat tgcggggggt gccgcaatga gctgttgcgt
     1981 accccccttt tttaagttgt tgatttttaa gtctttcgca tttcgcccta tatctagttc
     2041 tttggtgccc aaagaagggc acccctgcgg ggttccccca cgccttcggc gcggctcccc
     2101 ctccggcaaa aagtggcccc tccggggctt gttgatcgac tgcgcggcct tcggccttgc
     2161 ccaaggtggc gctgccccct tggaaccccc gcactcgccg ccgtgaggct cggggggcag
     2221 gcgggcgggc ttcgcccttc gactgccccc actcgcatag gcttgggtcg ttccaggcgc
     2281 gtcaaggcca agccgctgcg cggtcgctgc gcgagccttg acccgccttc cacttggtgt
     2341 ccaaccggca agcgaagcgc gcaggccgca ggccggaggc ttttccccag agaaaattaa
     2401 aaaaattgat ggggcaaggc cgcaggccgc gcagttggag ccggtgggta tgtggtcgaa
     2461 ggctgggtag ccggtgggca atccctgtgg tcaagctcgt gggcaggcgc agcctgtcca
     2521 tcagcttgtc cagcagggtt gtccacgggc cgagcgaagc gagccagccg gtggccgctc
     2581 gcggccatcg tccacatatc cacgggctgg caagggagcg cagcgaccgc gcagggcgaa
     2641 gcccggagag caagcccgta gggggggcgc gcccagctgt ctagggcggc ggatttgtcc
     2701 tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact
     2761 gagcctttcg ttttatttga tgcctttaat taagaagacg gcgacgaccc cgcgggccgc
     2821 cggcaacgcc ctgggtcttc actagtcttg gactcctgtt gatagatcca gtaatgacc
//
