LOCUS       sequence-146161-        4650 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mRFP1 expressed behind an ATc-inducible construct.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4650)
  AUTHORS   Florea M, Hagemann H, Santosa G, Abbott J, Micklem CN, 
            Spencer-Milnes X, de Arroyo Garcia L, Paschou D, Lazenbatt C, Kong 
            D, Chughtai H, Jensen K, Freemont PS, Kitney R, Reeve B, Ellis T
  TITLE     Engineering control of bacterial cellulose production using a 
            genetic toolkit and a new cellulose-producing strain.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Jun 14;113(24):E3431-40. doi: 
            10.1073/pnas.1522985113. Epub 2016 May 31.
  PUBMED    27247386
REFERENCE   2  (bases 1 to 4650)
  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: 
            "10.1073/pnas.1522985113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-06-14- 14"; volume: "113"; issue: "24"; pages: 
            "E3431-40"
COMMENT     Sequence Label: pTet02
FEATURES             Location/Qualifiers
     source          1..4650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             45..56
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             63..686
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(136..155)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      706..777
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      793..820
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    835..853
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    860..878
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             897..908
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             915..1592
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1637..1708
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1724..1751
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1783..1877
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        1925..2027
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2028..2687
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2094..2113)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     oriT            2836..2944
                     /note="incP origin of transfer"
     CDS             complement(2955..3617)
                     /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      3618..4389
                     /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(4504..4590)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4605..4627
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    4629..4650
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 tggtgcaaaa cctttgcggt atggcatgat agcgcctact agagaaagag gagaaatact
       61 agatgtccag attagataaa agtaaagtga ttaacagcgc attagagctg cttaatgagg
      121 tcggaatcga aggtttaaca acccgtaaac tcgcccagaa gctaggtgta gagcagccta
      181 cattgtattg gcatgtaaaa aataagcggg ctttgctcga cgccttagcc attgagatgt
      241 tagataggca ccatactcac ttttgccctt tagaagggga aagctggcaa gattttttac
      301 gtaataacgc taaaagtttt agatgtgctt tactaagtca tcgcgatgga gcaaaagtac
      361 atttaggtac acggcctaca gaaaaacagt atgaaactct cgaaaatcaa ttagcctttt
      421 tatgccaaca aggtttttca ctagagaatg cattatatgc actcagcgct gtggggcatt
      481 ttactttagg ttgcgtattg gaagatcaag agcatcaagt cgctaaagaa gaaagggaaa
      541 cacctactac tgatagtatg ccgccattat tacgacaagc tatcgaatta tttgatcacc
      601 aaggtgcaga gccagccttc ttattcggcc ttgaattgat catatgcgga ttagaaaaac
      661 aacttaaatg tgaaagtggg tcctaataat actagagcca ggcatcaaat aaaacgaaag
      721 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac
      781 tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttatatact agagtcccta
      841 tcagtgatag agattgacat ccctatcagt gatagagata ctgagcacta ctagtgaaag
      901 aggagaaata ctagatggct tcctccgaag acgttatcaa agagttcatg cgtttcaaag
      961 ttcgtatgga aggttccgtt aacggtcacg agttcgaaat cgaaggtgaa ggtgaaggtc
     1021 gtccgtacga aggtacccag accgctaaac tgaaagttac caaaggtggt ccgctgccgt
     1081 tcgcttggga catcctgtcc ccgcagttcc agtacggttc caaagcttac gttaaacacc
     1141 cggctgacat cccggactac ctgaaactgt ccttcccgga aggtttcaaa tgggaacgtg
     1201 ttatgaactt cgaagacggt ggtgttgtta ccgttaccca ggactcctcc ctgcaagacg
     1261 gtgagttcat ctacaaagtt aaactgcgtg gtaccaactt cccgtccgac ggtccggtta
     1321 tgcagaaaaa aaccatgggt tgggaagctt ccaccgaacg tatgtacccg gaagacggtg
     1381 ctctgaaagg tgaaatcaaa atgcgtctga aactgaaaga cggtggtcac tacgacgctg
     1441 aagttaaaac cacctacatg gctaaaaaac cggttcagct gccgggtgct tacaaaaccg
     1501 acatcaaact ggacatcacc tcccacaacg aagactacac catcgttgaa cagtacgaac
     1561 gtgctgaagg tcgtcactcc accggtgctt aataacgctg atagtgctag tgtagatcgc
     1621 tactagagcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     1681 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
     1741 gcctttctgc gtttatatac tagaagcggc cgctgcagct tggactcctg ttgatagatc
     1801 cagtaatgac ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt
     1861 tttttattgg tgagaatcca ggggtcccca ataattacga tttaaattgg cgaaaatgag
     1921 acgttgatcg gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg
     1981 gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
     2041 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
     2101 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
     2161 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
     2221 gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag ctggtgatat
     2281 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc
     2341 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg
     2401 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg
     2461 tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca
     2521 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga
     2581 tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc
     2641 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt gacttttgtc
     2701 cttttccgct gcataaccct gcttcggggt cattatagcg attttttcgg tatatccatc
     2761 ctttttcgca cgatatacag gattttgcca aagggttcgt gtagactttc cttggtgtat
     2821 ccaacggcgt cagccgggca ggataggtga agtaggccca cccgcgagcg ggtgttcctt
     2881 cttcactgtc ccttattcgc acctggcggt gctcaacggg aatcctgctc tgcgaggctg
     2941 gccgtaggcc ggccctaccg gcgcggcagc gttacccgtg tcggcggctc caacggctcg
     3001 ccatcgtcca gaaaacacgg ctcatcgggc atcggcaggc gctgctgccc gcgccgttcc
     3061 cattcctccg tttcggtcaa ggctggcagg tctggttcca tgcccggaat gccgggctgg
     3121 ctgggcggct cctcgccggg gccggtcggt agttgctgct cgcccggata cagggtcggg
     3181 atgcggcgca ggtcgccatg ccccaacagc gattcgtcct ggtcgtcgtg atcaaccacc
     3241 acggcggcac tgaacaccga caggcgcaac tggtcgcggg gctggcccca cgccacgcgg
     3301 tcattgacca cgtaggccga cacggtgccg gggccgttga gcttcacgac ggagatccag
     3361 cgctcggcca ccaagtcctt gactgcgtat tggaccgtcc gcaaagaacg tccgatgagc
     3421 ttggaaagtg tcttctggct gaccaccacg gcgttctggt ggcccatctg cgccacgagg
     3481 tgatgcagca gcattgccgc cgtgggtttc ctcgcaataa gcccggccca cgcctcatgc
     3541 gctttgcgtt ccgtttgcac ccagtgaccg ggcttgttct tggcttgaat gccgatttct
     3601 ctggactgcg tggccatgct tatctccatg cggtaggggt gccgcacggt tgcggcacca
     3661 tgcgcaatca gctgcaactt ttcggcagcg cgacaacaat tatgcgttgc gtaaaagtgg
     3721 cagtcaatta cagattttct ttaacctacg caatgagcta ttgcgggggg tgccgcaatg
     3781 agctgttgcg tacccccctt ttttaagttg ttgattttta agtctttcgc atttcgccct
     3841 atatctagtt ctttggtgcc caaagaaggg cacccctgcg gggttccccc acgccttcgg
     3901 cgcggctccc cctccggcaa aaagtggccc ctccggggct tgttgatcga ctgcgcggcc
     3961 ttcggccttg cccaaggtgg cgctgccccc ttggaacccc cgcactcgcc gccgtgaggc
     4021 tcggggggca ggcgggcggg cttcgccctt cgactgcccc cactcgcata ggcttgggtc
     4081 gttccaggcg cgtcaaggcc aagccgctgc gcggtcgctg cgcgagcctt gacccgcctt
     4141 ccacttggtg tccaaccggc aagcgaagcg cgcaggccgc aggccggagg cttttcccca
     4201 gagaaaatta aaaaaattga tggggcaagg ccgcaggccg cgcagttgga gccggtgggt
     4261 atgtggtcga aggctgggta gccggtgggc aatccctgtg gtcaagctcg tgggcaggcg
     4321 cagcctgtcc atcagcttgt ccagcagggt tgtccacggg ccgagcgaag cgagccagcc
     4381 ggtggccgct cgcggccatc gtccacatat ccacgggctg gcaagggagc gcagcgaccg
     4441 cgcagggcga agcccggaga gcaagcccgt agggggggcg cgcccagctg tctagggcgg
     4501 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     4561 gtctttcgac tgagcctttc gttttatttg atgcctttaa ttaaagcgga taacaatttc
     4621 acacaggaga attcgcggcc gcttctagag
//
