LOCUS       sequence-359535-        3890 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone for D1.2 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_137
FEATURES             Location/Qualifiers
     source          1..3890
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(290..376)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             562..573
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             580..1293
                     /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      1305..1376
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1392..1419
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1459..1553
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        1601..1703
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1704..2363
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1770..1789)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     oriT            2512..2620
                     /note="incP origin of transfer"
     CDS             complement(2631..3293)
                     /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(3294..3890,1..175)
                     /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 aattgatggg gcaaggccgc aggccgcgca gttggagccg gtgggtatgt ggtcgaaggc
       61 tgggtagccg gtgggcaatc cctgtggtca agctcgtggg caggcgcagc ctgtccatca
      121 gcttgtccag cagggttgtc cacgggccga gcgaagcgag ccagccggtg gccgctcgcg
      181 gccatcgtcc acatatccac gggctggcaa gggagcgcag cgaccgcgca gggcgaagcc
      241 cggagagcaa gcccgtaggg ggggcgcgcc cagctgtcta gggcggcgga tttgtcctac
      301 tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct ttcgactgag
      361 cctttcgttt tatttgatgc ctttaattaa gaagacggcc ctgacctgag accgaaagtg
      421 aaacgtgatt tcatgcgtca ttttgaacat tttgtaaatc ttatttaata atgtgtgcgg
      481 caattcacat ttaatttatg aatgttttct taacatcgcg gcaactcaag aaacggcagg
      541 ttcggatctt agctactaga gaaagaggag aaatactaga tgcgtaaagg cgaagagctg
      601 ttcactggtg tcgtccctat tctggtggaa ctggatggtg atgtcaacgg tcataagttt
      661 tccgtgcgtg gcgagggtga aggtgacgca actaatggta aactgacgct gaagttcatc
      721 tgtactactg gtaaactgcc ggttccttgg ccgactctgg taacgacgct gacttatggt
      781 gttcagtgct ttgctcgtta tccggaccat atgaagcagc atgacttctt caagtccgcc
      841 atgccggaag gctatgtgca ggaacgcacg atttccttta aggatgacgg cacgtacaaa
      901 acgcgtgcgg aagtgaaatt tgaaggcgat accctggtaa accgcattga gctgaaaggc
      961 attgacttta aagaggacgg caatatcctg ggccataagc tggaatacaa ttttaacagc
     1021 cacaatgttt acatcaccgc cgataaacaa aaaaatggca ttaaagcgaa ttttaaaatt
     1081 cgccacaacg tggaggatgg cagcgtgcag ctggctgatc actaccagca aaacactcca
     1141 atcggtgatg gtcctgttct gctgccagac aatcactatc tgagcacgca aagcgttctg
     1201 tctaaagatc cgaacgagaa acgcgatcat atggttctgc tggagttcgt aaccgcagcg
     1261 ggcatcacgc atggtatgga tgaactgtac aaatgaccag gcatcaaata aaacgaaagg
     1321 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact
     1381 agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttataggtct caaacgccga
     1441 gggtcttcac tagtcttgga ctcctgttga tagatccagt aatgacctca gaactccatc
     1501 tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccagggg
     1561 tccccaataa ttacgattta aattggcgaa aatgagacgt tgatcggcac gtaagaggtt
     1621 ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat
     1681 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
     1741 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
     1801 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
     1861 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
     1921 ttcgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
     1981 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
     2041 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
     2101 atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt
     2161 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
     2221 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
     2281 atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
     2341 atgagtggca gggcggggcg taatttgact tttgtccttt tccgctgcat aaccctgctt
     2401 cggggtcatt atagcgattt tttcggtata tccatccttt ttcgcacgat atacaggatt
     2461 ttgccaaagg gttcgtgtag actttccttg gtgtatccaa cggcgtcagc cgggcaggat
     2521 aggtgaagta ggcccacccg cgagcgggtg ttccttcttc actgtccctt attcgcacct
     2581 ggcggtgctc aacgggaatc ctgctctgcg aggctggccg taggccggcc ctaccggcgc
     2641 ggcagcgtta cccgtgtcgg cggctccaac ggctcgccat cgtccagaaa acacggctca
     2701 tcgggcatcg gcaggcgctg ctgcccgcgc cgttcccatt cctccgtttc ggtcaaggct
     2761 ggcaggtctg gttccatgcc cggaatgccg ggctggctgg gcggctcctc gccggggccg
     2821 gtcggtagtt gctgctcgcc cggatacagg gtcgggatgc ggcgcaggtc gccatgcccc
     2881 aacagcgatt cgtcctggtc gtcgtgatca accaccacgg cggcactgaa caccgacagg
     2941 cgcaactggt cgcggggctg gccccacgcc acgcggtcat tgaccacgta ggccgacacg
     3001 gtgccggggc cgttgagctt cacgacggag atccagcgct cggccaccaa gtccttgact
     3061 gcgtattgga ccgtccgcaa agaacgtccg atgagcttgg aaagtgtttt ctggctgacc
     3121 accacggcgt tctggtggcc catctgcgcc acgaggtgat gcagcagcat tgccgccgtg
     3181 ggtttcctcg caataagccc ggcccacgcc tcatgcgctt tgcgttccgt ttgcacccag
     3241 tgaccgggct tgttcttggc ttgaatgccg atttctctgg actgcgtggc catgcttatc
     3301 tccatgcggt aggggtgccg cacggttgcg gcaccatgcg caatcagctg caacttttcg
     3361 gcagcgcgac aacaattatg cgttgcgtaa aagtggcagt caattacaga ttttctttaa
     3421 cctacgcaat gagctattgc ggggggtgcc gcaatgagct gttgcgtacc cccctttttt
     3481 aagttgttga tttttaagtc tttcgcattt cgccctatat ctagttcttt ggtgcccaaa
     3541 gaagggcacc cctgcggggt tcccccacgc cttcggcgcg gctccccctc cggcaaaaag
     3601 tggcccctcc ggggcttgtt gatcgactgc gcggccttcg gccttgccca aggtggcgct
     3661 gcccccttgg aacccccgca ctcgccgccg tgaggctcgg ggggcaggcg ggcgggcttc
     3721 gcccttcgac tgcccccact cgcataggct tgggtcgttc caggcgcgtc aaggccaagc
     3781 cgctgcgcgg tcgctgcgcg agccttgacc cgccttccac ttggtgtcca accggcaagc
     3841 gaagcgcgca ggccgcaggc cggaggcttt tccccagaga aaattaaaaa
//
