LOCUS       sequence-146148-        3797 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mRFP1 behind constitutive promoter J23101 in pSEVA331Bb backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3797)
  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 3797)
  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: J23101-mRFP1-331Bb
FEATURES             Location/Qualifiers
     source          1..3797
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..35
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     RBS             44..55
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             62..739
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      784..855
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      871..898
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      930..1024
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        1072..1174
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1175..1834
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1241..1260)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     oriT            1983..2091
                     /note="incP origin of transfer"
     CDS             complement(2102..2764)
                     /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      2765..3536
                     /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(3651..3737)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     3752..3774
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    3776..3797
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 tttacagcta gctcagtcct aggtattatg ctagctacta gtgaaagagg agaaatacta
       61 gatggcttcc tccgaagacg ttatcaaaga gttcatgcgt ttcaaagttc gtatggaagg
      121 ttccgttaac ggtcacgagt tcgaaatcga aggtgaaggt gaaggtcgtc cgtacgaagg
      181 tacccagacc gctaaactga aagttaccaa aggtggtccg ctgccgttcg cttgggacat
      241 cctgtccccg cagttccagt acggttccaa agcttacgtt aaacacccgg ctgacatccc
      301 ggactacctg aaactgtcct tcccggaagg tttcaaatgg gaacgtgtta tgaacttcga
      361 agacggtggt gttgttaccg ttacccagga ctcctccctg caagacggtg agttcatcta
      421 caaagttaaa ctgcgtggta ccaacttccc gtccgacggt ccggttatgc agaaaaaaac
      481 catgggttgg gaagcttcca ccgaacgtat gtacccggaa gacggtgctc tgaaaggtga
      541 aatcaaaatg cgtctgaaac tgaaagacgg tggtcactac gacgctgaag ttaaaaccac
      601 ctacatggct aaaaaaccgg ttcagctgcc gggtgcttac aaaaccgaca tcaaactgga
      661 catcacctcc cacaacgaag actacaccat cgttgaacag tacgaacgtg ctgaaggtcg
      721 tcactccacc ggtgcttaat aacgctgata gtgctagtgt agatcgctac tagagccagg
      781 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
      841 tcggtgaacg ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt
      901 tatatactag aagcggccgc tgcagcttgg actcctgttg atagatccag taatgacctc
      961 agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga
     1021 gaatccaggg gtccccaata attacgattt aaattggcga aaatgagacg ttgatcggca
     1081 cgtaagaggt tccaactttc accataatga aataagatca ctaccgggcg tattttttga
     1141 gttatcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac
     1201 caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc
     1261 tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa
     1321 gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc
     1381 tcatccggaa tttcgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca
     1441 cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata
     1501 ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga
     1561 aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc
     1621 ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc
     1681 cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat
     1741 tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca
     1801 acagtactgc gatgagtggc agggcggggc gtaatttgac ttttgtcctt ttccgctgca
     1861 taaccctgct tcggggtcat tatagcgatt ttttcggtat atccatcctt tttcgcacga
     1921 tatacaggat tttgccaaag ggttcgtgta gactttcctt ggtgtatcca acggcgtcag
     1981 ccgggcagga taggtgaagt aggcccaccc gcgagcgggt gttccttctt cactgtccct
     2041 tattcgcacc tggcggtgct caacgggaat cctgctctgc gaggctggcc gtaggccggc
     2101 cctaccggcg cggcagcgtt acccgtgtcg gcggctccaa cggctcgcca tcgtccagaa
     2161 aacacggctc atcgggcatc ggcaggcgct gctgcccgcg ccgttcccat tcctccgttt
     2221 cggtcaaggc tggcaggtct ggttccatgc ccggaatgcc gggctggctg ggcggctcct
     2281 cgccggggcc ggtcggtagt tgctgctcgc ccggatacag ggtcgggatg cggcgcaggt
     2341 cgccatgccc caacagcgat tcgtcctggt cgtcgtgatc aaccaccacg gcggcactga
     2401 acaccgacag gcgcaactgg tcgcggggct ggccccacgc cacgcggtca ttgaccacgt
     2461 aggccgacac ggtgccgggg ccgttgagct tcacgacgga gatccagcgc tcggccacca
     2521 agtccttgac tgcgtattgg accgtccgca aagaacgtcc gatgagcttg gaaagtgtct
     2581 tctggctgac caccacggcg ttctggtggc ccatctgcgc cacgaggtga tgcagcagca
     2641 ttgccgccgt gggtttcctc gcaataagcc cggcccacgc ctcatgcgct ttgcgttccg
     2701 tttgcaccca gtgaccgggc ttgttcttgg cttgaatgcc gatttctctg gactgcgtgg
     2761 ccatgcttat ctccatgcgg taggggtgcc gcacggttgc ggcaccatgc gcaatcagct
     2821 gcaacttttc ggcagcgcga caacaattat gcgttgcgta aaagtggcag tcaattacag
     2881 attttcttta acctacgcaa tgagctattg cggggggtgc cgcaatgagc tgttgcgtac
     2941 cccccttttt taagttgttg atttttaagt ctttcgcatt tcgccctata tctagttctt
     3001 tggtgcccaa agaagggcac ccctgcgggg ttcccccacg ccttcggcgc ggctccccct
     3061 ccggcaaaaa gtggcccctc cggggcttgt tgatcgactg cgcggccttc ggccttgccc
     3121 aaggtggcgc tgcccccttg gaacccccgc actcgccgcc gtgaggctcg gggggcaggc
     3181 gggcgggctt cgcccttcga ctgcccccac tcgcataggc ttgggtcgtt ccaggcgcgt
     3241 caaggccaag ccgctgcgcg gtcgctgcgc gagccttgac ccgccttcca cttggtgtcc
     3301 aaccggcaag cgaagcgcgc aggccgcagg ccggaggctt ttccccagag aaaattaaaa
     3361 aaattgatgg ggcaaggccg caggccgcgc agttggagcc ggtgggtatg tggtcgaagg
     3421 ctgggtagcc ggtgggcaat ccctgtggtc aagctcgtgg gcaggcgcag cctgtccatc
     3481 agcttgtcca gcagggttgt ccacgggccg agcgaagcga gccagccggt ggccgctcgc
     3541 ggccatcgtc cacatatcca cgggctggca agggagcgca gcgaccgcgc agggcgaagc
     3601 ccggagagca agcccgtagg gggggcgcgc ccagctgtct agggcggcgg atttgtccta
     3661 ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc tttcgactga
     3721 gcctttcgtt ttatttgatg cctttaatta aagcggataa caatttcaca caggagaatt
     3781 cgcggccgct tctagag
//
