LOCUS       sequence-358969-        2888 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone for creation of entry parts for the KTK, contains 
            sfGFP dropout.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2888)
  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 2888)
  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_001
FEATURES             Location/Qualifiers
     source          1..2888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     107..126
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     RBS             467..478
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             485..1198
                     /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      1210..1281
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1297..1324
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(1438..2229,2230..2298))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2061..2080
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2299..2403)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(2506..2888,1..206)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
       61 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
      121 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
      181 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
      241 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
      301 accgtagtcg gcgagacgga aagtgaaacg tgatttcatg cgtcattttg aacattttgt
      361 aaatcttatt taataatgtg tgcggcaatt cacatttaat ttatgaatgt tttcttaaca
      421 tcgcggcaac tcaagaaacg gcaggttcgg atcttagcta ctagagaaag aggagaaata
      481 ctagatgcgt aaaggcgaag agctgttcac tggtgtcgtc cctattctgg tggaactgga
      541 tggtgatgtc aacggtcata agttttccgt gcgtggcgag ggtgaaggtg acgcaactaa
      601 tggtaaactg acgctgaagt tcatctgtac tactggtaaa ctgccggttc cttggccgac
      661 tctggtaacg acgctgactt atggtgttca gtgctttgct cgttatccgg accatatgaa
      721 gcagcatgac ttcttcaagt ccgccatgcc ggaaggctat gtgcaggaac gcacgatttc
      781 ctttaaggat gacggcacgt acaaaacgcg tgcggaagtg aaatttgaag gcgataccct
      841 ggtaaaccgc attgagctga aaggcattga ctttaaagag gacggcaata tcctgggcca
      901 taagctggaa tacaatttta acagccacaa tgtttacatc accgccgata aacaaaaaaa
      961 tggcattaaa gcgaatttta aaattcgcca caacgtggag gatggcagcg tgcagctggc
     1021 tgatcactac cagcaaaaca ctccaatcgg tgatggtcct gttctgctgc cagacaatca
     1081 ctatctgagc acgcaaagcg ttctgtctaa agatccgaac gagaaacgcg atcatatggt
     1141 tctgctggag ttcgtaaccg cagcgggcat cacgcatggt atggatgaac tgtacaaatg
     1201 accaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     1261 tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc
     1321 tgcgtttata cgtctctgac caagattatc aaaaaggatc ttcacctaga tccttttaaa
     1381 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1441 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1501 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1561 tgctgcaatg ataccgcggg acccacgctc accggctcca gatttatcag caataaacca
     1621 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1681 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1741 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1801 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1861 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1921 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1981 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2041 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2101 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2161 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2221 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2281 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2341 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2401 gcgcacattt ccccgaaaag tgccacctgt catgaccaaa atcccttaac gtgagttttc
     2461 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     2521 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     2581 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2641 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     2701 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     2761 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     2821 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2881 atacctac
//
