LOCUS       sequence-146160-        4704 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 4704)
  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 4704)
  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: pTet01
FEATURES             Location/Qualifiers
     source          1..4704
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..35
                     /label=BBa_J23118
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.56"
     CDS             61..681
                     /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(134..153)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             682..714
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag (LVA)
                     /note="mutant LVA variant that confers accelerated 
                     degradation under some conditions (Andersen et al., 1998)"
                     /translation="AANDENYALVA"
     terminator      762..833
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      849..876
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    891..909
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    916..934
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             951..962
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             969..1646
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1691..1762
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1778..1805
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1837..1931
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        1979..2081
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2082..2741
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2148..2167)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     oriT            2890..2998
                     /note="incP origin of transfer"
     CDS             complement(3009..3671)
                     /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      3672..4443
                     /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(4558..4644)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4659..4681
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    4683..4704
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ttgacggcta gctcagtcct aggtattgtg ctagctacta gagtcacaca ggactactag
       61 atgtccagat tagataaaag taaagtgatt aacagcgcat tagagctgct taatgaggtc
      121 ggaatcgaag gtttaacaac ccgtaaactc gcccagaagc taggtgtaga gcagcctaca
      181 ttgtattggc atgtaaaaaa taagcgggct ttgctcgacg ccttagccat tgagatgtta
      241 gataggcacc atactcactt ttgcccttta gaaggggaaa gctggcaaga ttttttacgt
      301 aataacgcta aaagttttag atgtgcttta ctaagtcatc gcgatggagc aaaagtacat
      361 ttaggtacac ggcctacaga aaaacagtat gaaactctcg aaaatcaatt agccttttta
      421 tgccaacaag gtttttcact agagaatgca ttatatgcac tcagcgctgt ggggcatttt
      481 actttaggtt gcgtattgga agatcaagag catcaagtcg ctaaagaaga aagggaaaca
      541 cctactactg atagtatgcc gccattatta cgacaagcta tcgaattatt tgatcaccaa
      601 ggtgcagagc cagccttctt attcggcctt gaattgatca tatgcggatt agaaaaacaa
      661 cttaaatgtg aaagtgggtc cgctgcaaac gacgaaaact acgctttagt agcttaataa
      721 cactgatagt gctagtgtag atcactacta gagccaggca tcaaataaaa cgaaaggctc
      781 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga
      841 gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta tatactagag tccctatcag
      901 tgatagagat tgacatccct atcagtgata gagatactga gcactactag aaagaggaga
      961 aatactagat ggcttcctcc gaagacgtta tcaaagagtt catgcgtttc aaagttcgta
     1021 tggaaggttc cgttaacggt cacgagttcg aaatcgaagg tgaaggtgaa ggtcgtccgt
     1081 acgaaggtac ccagaccgct aaactgaaag ttaccaaagg tggtccgctg ccgttcgctt
     1141 gggacatcct gtccccgcag ttccagtacg gttccaaagc ttacgttaaa cacccggctg
     1201 acatcccgga ctacctgaaa ctgtccttcc cggaaggttt caaatgggaa cgtgttatga
     1261 acttcgaaga cggtggtgtt gttaccgtta cccaggactc ctccctgcaa gacggtgagt
     1321 tcatctacaa agttaaactg cgtggtacca acttcccgtc cgacggtccg gttatgcaga
     1381 aaaaaaccat gggttgggaa gcttccaccg aacgtatgta cccggaagac ggtgctctga
     1441 aaggtgaaat caaaatgcgt ctgaaactga aagacggtgg tcactacgac gctgaagtta
     1501 aaaccaccta catggctaaa aaaccggttc agctgccggg tgcttacaaa accgacatca
     1561 aactggacat cacctcccac aacgaagact acaccatcgt tgaacagtac gaacgtgctg
     1621 aaggtcgtca ctccaccggt gcttaataac gctgatagtg ctagtgtaga tcgctactag
     1681 agccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     1741 ttgtttgtcg gtgaacgctc tctactagag tcacactggc tcaccttcgg gtgggccttt
     1801 ctgcgtttat atactagaag cggccgctgc agcttggact cctgttgata gatccagtaa
     1861 tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta
     1921 ttggtgagaa tccaggggtc cccaataatt acgatttaaa ttggcgaaaa tgagacgttg
     1981 atcggcacgt aagaggttcc aactttcacc ataatgaaat aagatcacta ccgggcgtat
     2041 tttttgagtt atcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg
     2101 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt
     2161 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga
     2221 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga
     2281 tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg atatgggata
     2341 gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga
     2401 gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt
     2461 acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag
     2521 ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct
     2581 tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc
     2641 tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg
     2701 aattacaaca gtactgcgat gagtggcagg gcggggcgta atttgacttt tgtccttttc
     2761 cgctgcataa ccctgcttcg gggtcattat agcgattttt tcggtatatc catccttttt
     2821 cgcacgatat acaggatttt gccaaagggt tcgtgtagac tttccttggt gtatccaacg
     2881 gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt ccttcttcac
     2941 tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag gctggccgta
     3001 ggccggccct accggcgcgg cagcgttacc cgtgtcggcg gctccaacgg ctcgccatcg
     3061 tccagaaaac acggctcatc gggcatcggc aggcgctgct gcccgcgccg ttcccattcc
     3121 tccgtttcgg tcaaggctgg caggtctggt tccatgcccg gaatgccggg ctggctgggc
     3181 ggctcctcgc cggggccggt cggtagttgc tgctcgcccg gatacagggt cgggatgcgg
     3241 cgcaggtcgc catgccccaa cagcgattcg tcctggtcgt cgtgatcaac caccacggcg
     3301 gcactgaaca ccgacaggcg caactggtcg cggggctggc cccacgccac gcggtcattg
     3361 accacgtagg ccgacacggt gccggggccg ttgagcttca cgacggagat ccagcgctcg
     3421 gccaccaagt ccttgactgc gtattggacc gtccgcaaag aacgtccgat gagcttggaa
     3481 agtgtcttct ggctgaccac cacggcgttc tggtggccca tctgcgccac gaggtgatgc
     3541 agcagcattg ccgccgtggg tttcctcgca ataagcccgg cccacgcctc atgcgctttg
     3601 cgttccgttt gcacccagtg accgggcttg ttcttggctt gaatgccgat ttctctggac
     3661 tgcgtggcca tgcttatctc catgcggtag gggtgccgca cggttgcggc accatgcgca
     3721 atcagctgca acttttcggc agcgcgacaa caattatgcg ttgcgtaaaa gtggcagtca
     3781 attacagatt ttctttaacc tacgcaatga gctattgcgg ggggtgccgc aatgagctgt
     3841 tgcgtacccc ccttttttaa gttgttgatt tttaagtctt tcgcatttcg ccctatatct
     3901 agttctttgg tgcccaaaga agggcacccc tgcggggttc ccccacgcct tcggcgcggc
     3961 tccccctccg gcaaaaagtg gcccctccgg ggcttgttga tcgactgcgc ggccttcggc
     4021 cttgcccaag gtggcgctgc ccccttggaa cccccgcact cgccgccgtg aggctcgggg
     4081 ggcaggcggg cgggcttcgc ccttcgactg cccccactcg cataggcttg ggtcgttcca
     4141 ggcgcgtcaa ggccaagccg ctgcgcggtc gctgcgcgag ccttgacccg ccttccactt
     4201 ggtgtccaac cggcaagcga agcgcgcagg ccgcaggccg gaggcttttc cccagagaaa
     4261 attaaaaaaa ttgatggggc aaggccgcag gccgcgcagt tggagccggt gggtatgtgg
     4321 tcgaaggctg ggtagccggt gggcaatccc tgtggtcaag ctcgtgggca ggcgcagcct
     4381 gtccatcagc ttgtccagca gggttgtcca cgggccgagc gaagcgagcc agccggtggc
     4441 cgctcgcggc catcgtccac atatccacgg gctggcaagg gagcgcagcg accgcgcagg
     4501 gcgaagcccg gagagcaagc ccgtaggggg ggcgcgccca gctgtctagg gcggcggatt
     4561 tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttt
     4621 cgactgagcc tttcgtttta tttgatgcct ttaattaaag cggataacaa tttcacacag
     4681 gagaattcgc ggccgcttct agag
//
