LOCUS       sequence-455853-        2876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2876)
  AUTHORS   Guet CC, Bruneaux L, Min TL, Siegal-Gaskins D, Figueroa I, Emonet T,
            Cluzel P
  TITLE     Minimally invasive determination of mRNA concentration in single 
            living bacteria.
  JOURNAL   Nucleic Acids Res. 2008 Jul . 36(12):e73.
  PUBMED    18515347
REFERENCE   2  (bases 1 to 2876)
  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"; journalName: "Nucleic 
            Acids Res. 2008 Jul . 36(12):e73."
COMMENT     Sequence Label: pZE31-DsRed-ms2x2
FEATURES             Location/Qualifiers
     source          1..2876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(6..24,25..30,31..47,48..53,54..79)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    6..24
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    31..49
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             89..100
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             109..786
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDNTENVIKEFMRFKVRMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYTKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLEGGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTVVEQYERTEARHH
                     LFL"
     primer_bind     complement(289..309)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     507..526
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     701..724
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     795..811
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     795..811
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_RNA        851..869
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        890..908
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     terminator      973..1059
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1224..1812)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1304..1323)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1900..1994)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2015..2674)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2589..2608
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2675..2777)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     2843..2861
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgagtccct atcagtgata gagattgaca tccctatcag tgatagagat actgagcaca
       61 tcagcaggac gcactgaccg aattcattaa agaggagaaa ggtaccgcat ggacaacacc
      121 gagaacgtca tcaaggagtt catgcgcttc aaggtgcgca tggagggctc cgtgaacggc
      181 cacgagttcg agatcgaggg cgagggcgag ggcaagccct acgagggcac ccagaccgcc
      241 aagctgcagg tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag
      301 ttccagtacg gctccaaggt gtacaccaag caccccgccg acatccccga ctacaagaag
      361 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
      421 gtgaccgtga cccaggactc ctccctgcag gacggcacct tcatctacca cgtgaagttc
      481 atcggcgtga acttcccctc cgacggcccc gtaatgcaga agaagactct gggctgggag
      541 ccctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccacaaggcc
      601 ctgaagctgg agggcggcgg ccactacctg gtggagttca agtccatcta catggccaag
      661 aagcccgtgc agctgcccgg ctactactac gtggactcca agctggacat cacctcccac
      721 aacgaggact acaccgtggt ggagcagtac gagcgcaccg aggcccgcca ccacctgttc
      781 ctgtaagctt ccgctctaga actagtggat cccccgggca gcttgcatgc ctgcaggtcg
      841 actctagaaa acatgaggat cacccatgtc tgcaggtcga ctctagaaaa catgaggatc
      901 acccatgtct gcaggtcgac tctagaggat cggaattcga tatcaagctt aattagctga
      961 gtctagaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     1021 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgcccta gacctagggc
     1081 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1141 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1201 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1261 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1321 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1381 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1441 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1501 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1561 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1621 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     1681 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1741 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1801 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1861 aactcacgtt aagggatttt ggtcatgact agtgcttgga ttctcaccaa taaaaaacgc
     1921 ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc
     1981 tatcaacagg agtccaagcg agctcgatat caaattacgc cccgccctgc cactcatcgc
     2041 agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat
     2101 gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg
     2161 tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac
     2221 tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg
     2281 ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat
     2341 cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt
     2401 aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc atacgaaatt
     2461 ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct
     2521 tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg
     2581 tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat
     2641 caacggtggt atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc
     2701 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac
     2761 ctcttacgtg ccgatcaacg tctcattttc gccagatatc gacgtctaag aaaccattat
     2821 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcacc
//
