LOCUS       sequence-359557-        3890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone for D1.1 cloning, contains sfGFP dropout.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3890)
  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 3890)
  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_136
FEATURES             Location/Qualifiers
     source          1..3890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      239..1010
                     /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(1125..1211)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             1397..1408
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1415..2128
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2140..2211
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2227..2254
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      2294..2388
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        2436..2538
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2539..3198
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2605..2624)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     oriT            3347..3455
                     /note="incP origin of transfer"
     CDS             complement(join(3466..3890,1..238))
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
ORIGIN
        1 tgactgcgta ttggaccgtc cgcaaagaac gtccgatgag cttggaaagt gttttctggc
       61 tgaccaccac ggcgttctgg tggcccatct gcgccacgag gtgatgcagc agcattgccg
      121 ccgtgggttt cctcgcaata agcccggccc acgcctcatg cgctttgcgt tccgtttgca
      181 cccagtgacc gggcttgttc ttggcttgaa tgccgatttc tctggactgc gtggccatgc
      241 ttatctccat gcggtagggg tgccgcacgg ttgcggcacc atgcgcaatc agctgcaact
      301 tttcggcagc gcgacaacaa ttatgcgttg cgtaaaagtg gcagtcaatt acagattttc
      361 tttaacctac gcaatgagct attgcggggg gtgccgcaat gagctgttgc gtacccccct
      421 tttttaagtt gttgattttt aagtctttcg catttcgccc tatatctagt tctttggtgc
      481 ccaaagaagg gcacccctgc ggggttcccc cacgccttcg gcgcggctcc ccctccggca
      541 aaaagtggcc cctccggggc ttgttgatcg actgcgcggc cttcggcctt gcccaaggtg
      601 gcgctgcccc cttggaaccc ccgcactcgc cgccgtgagg ctcggggggc aggcgggcgg
      661 gcttcgccct tcgactgccc ccactcgcat aggcttgggt cgttccaggc gcgtcaaggc
      721 caagccgctg cgcggtcgct gcgcgagcct tgacccgcct tccacttggt gtccaaccgg
      781 caagcgaagc gcgcaggccg caggccggag gcttttcccc agagaaaatt aaaaaaattg
      841 atggggcaag gccgcaggcc gcgcagttgg agccggtggg tatgtggtcg aaggctgggt
      901 agccggtggg caatccctgt ggtcaagctc gtgggcaggc gcagcctgtc catcagcttg
      961 tccagcaggg ttgtccacgg gccgagcgaa gcgagccagc cggtggccgc tcgcggccat
     1021 cgtccacata tccacgggct ggcaagggag cgcagcgacc gcgcagggcg aagcccggag
     1081 agcaagcccg tagggggggc gcgcccagct gtctagggcg gcggatttgt cctactcagg
     1141 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt
     1201 cgttttattt gatgccttta attaagaaga cggcgacgac ctgagaccga aagtgaaacg
     1261 tgatttcatg cgtcattttg aacattttgt aaatcttatt taataatgtg tgcggcaatt
     1321 cacatttaat ttatgaatgt tttcttaaca tcgcggcaac tcaagaaacg gcaggttcgg
     1381 atcttagcta ctagagaaag aggagaaata ctagatgcgt aaaggcgaag agctgttcac
     1441 tggtgtcgtc cctattctgg tggaactgga tggtgatgtc aacggtcata agttttccgt
     1501 gcgtggcgag ggtgaaggtg acgcaactaa tggtaaactg acgctgaagt tcatctgtac
     1561 tactggtaaa ctgccggttc cttggccgac tctggtaacg acgctgactt atggtgttca
     1621 gtgctttgct cgttatccgg accatatgaa gcagcatgac ttcttcaagt ccgccatgcc
     1681 ggaaggctat gtgcaggaac gcacgatttc ctttaaggat gacggcacgt acaaaacgcg
     1741 tgcggaagtg aaatttgaag gcgataccct ggtaaaccgc attgagctga aaggcattga
     1801 ctttaaagag gacggcaata tcctgggcca taagctggaa tacaatttta acagccacaa
     1861 tgtttacatc accgccgata aacaaaaaaa tggcattaaa gcgaatttta aaattcgcca
     1921 caacgtggag gatggcagcg tgcagctggc tgatcactac cagcaaaaca ctccaatcgg
     1981 tgatggtcct gttctgctgc cagacaatca ctatctgagc acgcaaagcg ttctgtctaa
     2041 agatccgaac gagaaacgcg atcatatggt tctgctggag ttcgtaaccg cagcgggcat
     2101 cacgcatggt atggatgaac tgtacaaatg accaggcatc aaataaaacg aaaggctcag
     2161 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt
     2221 cacactggct caccttcggg tgggcctttc tgcgtttata ggtctcaaac gccctgggtc
     2281 ttcactagtc ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat
     2341 ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aggggtcccc
     2401 aataattacg atttaaattg gcgaaaatga gacgttgatc ggcacgtaag aggttccaac
     2461 tttcaccata atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca
     2521 ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc
     2581 caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac
     2641 cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag
     2701 ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc ggaatttcgt
     2761 atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt
     2821 ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg
     2881 cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc
     2941 cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc
     3001 agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc
     3061 aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc
     3121 gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag
     3181 tggcagggcg gggcgtaatt tgacttttgt ccttttccgc tgcataaccc tgcttcgggg
     3241 tcattatagc gattttttcg gtatatccat cctttttcgc acgatataca ggattttgcc
     3301 aaagggttcg tgtagacttt ccttggtgta tccaacggcg tcagccgggc aggataggtg
     3361 aagtaggccc acccgcgagc gggtgttcct tcttcactgt cccttattcg cacctggcgg
     3421 tgctcaacgg gaatcctgct ctgcgaggct ggccgtaggc cggccctacc ggcgcggcag
     3481 cgttacccgt gtcggcggct ccaacggctc gccatcgtcc agaaaacacg gctcatcggg
     3541 catcggcagg cgctgctgcc cgcgccgttc ccattcctcc gtttcggtca aggctggcag
     3601 gtctggttcc atgcccggaa tgccgggctg gctgggcggc tcctcgccgg ggccggtcgg
     3661 tagttgctgc tcgcccggat acagggtcgg gatgcggcgc aggtcgccat gccccaacag
     3721 cgattcgtcc tggtcgtcgt gatcaaccac cacggcggca ctgaacaccg acaggcgcaa
     3781 ctggtcgcgg ggctggcccc acgccacgcg gtcattgacc acgtaggccg acacggtgcc
     3841 ggggccgttg agcttcacga cggagatcca gcgctcggcc accaagtcct
//
