LOCUS       sequence-378330-        3593 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RFP generator controlled by Plux2 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3593)
  AUTHORS   Liu Y, Pinto F, Wan X, Yang Z, Peng S, Li M, Cooper JM, Xie Z, 
            French CE, Wang B
  TITLE     Reprogrammed tracrRNAs enable repurposing of RNAs as crRNAs and 
            sequence-specific RNA biosensors.
  JOURNAL   Nat Commun. 2022 Apr 11;13(1):1937. doi: 10.1038/s41467-022-29604-x.
  PUBMED    35410423
REFERENCE   2  (bases 1 to 3593)
  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: "Nat 
            Commun."; date: "2022-04-11"; volume: "13(1):1937. doi"; pages: " 
            10.1038/s41467-022-29604-x"
COMMENT     Sequence Label: pLY185
FEATURES             Location/Qualifiers
     source          1..3593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(24..41)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(158..703)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(238..257)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(817..911)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(932..1591)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1506..1525
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1592..1694)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     RBS             2633..2644
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(2656..2690)
                     /label=BBa_J23117
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.06"
     terminator      2698..2725
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             2805..2816
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2823..3533
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2968..2986)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3236..3255
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 ggcctaggga tatattccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg
       61 cggcgagcgg aaatggctta cgaacggggc ggagatttcc tggaagatgc caggaagata
      121 cttaacaggg aagtgagagg gccgcggcaa agccgttttt ccataggctc cgcccccctg
      181 acaagcatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca ggactataaa
      241 gataccaggc gtttccccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt
      301 ttaccggtgt cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt
      361 tccgggtagg cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac
      421 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca
      481 ccactggcag cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg
      541 ttaaggctaa actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg
      601 gttcaaagag ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc
      661 gttttcagag caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattaa
      721 tcagataaaa tatttctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt
      781 cagccccata cgatataagt tgttactagt gcttggattc tcaccaataa aaaacgcccg
      841 gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat
      901 caacaggagt ccaagcgagc tcgatatcaa attacgcccc gccctgccac tcatcgcagt
      961 actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa
     1021 cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga
     1081 aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca
     1141 cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca
     1201 ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt
     1261 cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac
     1321 aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata cgaaattccg
     1381 gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat
     1441 ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac
     1501 attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa
     1561 cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg
     1621 ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc
     1681 ttacgtgccg atcaacgtct cattttcgcc agatatctaa ttcgcggccg cttctagagc
     1741 ctagttgtct tcatgcatga agacaaaatt aatactagag ggaccaaaac gaaaaaagac
     1801 gcttttcagc gtcttattgt tcgtctttgg taccgagctc tagtagtgat ctacactagc
     1861 actatcagtg ttattaattt ttaaagtatg ggcaatcaat tgctcctgtt aaaattgctt
     1921 tagaaatact ttggcagcgg tttgttgtat tgagtttcat ttgcgcattg gttaaatgga
     1981 aagtgacagt acgctcactg caacctaata tttttgaaat atcccaagag ctttttcctt
     2041 cgcatgccca cgctaaacat tctttttctc ttttggttaa atcgttgttt gatttattat
     2101 ttgctatatt tatttttcga taattatcaa ctagagaagg aacaattaat ggtatgttca
     2161 tacacgcatg taaaaataaa ctatctatat agttgtcttt ttctgaatgt gcaaaactaa
     2221 gcattccgaa gccattgtta gccgtatgaa tagggaaact aaacccagtg ataagacctg
     2281 atgttttcgc ttctttaatt acatttggag attttttatt tacagcattg ttttcaaata
     2341 tattccaatt aattggtgaa tgattggagt tagaataatc tactatagga tcatatttta
     2401 ttaaattagc gtcatcataa tattgcctcc attttttagg gtaattatct aggattgaaa
     2461 tatcagattt aaccatagaa tgaggataaa tgatcgcgag taaataatat tcacaatgta
     2521 ccattttagt catatcagat aagcattgat taatatcatt attgcttcta caagctttaa
     2581 ttttattaat tattctgtat gtgtcgtcgg catttatgtt tttcatctag tatttctcct
     2641 ctttaatctc tagtagctag cacaatccct aggactgagc tagctgtcaa tcacactggc
     2701 tcaccttcgg gtgggccttt ctgcgtttat atactagaga cctgtaggat cgtacaggtt
     2761 tacgcaagaa aatggtttgt tactttcgaa taaaactaga gattaaagag gagaaatact
     2821 agatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     2881 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     2941 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     3001 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     3061 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     3121 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tcacttcagg
     3181 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     3241 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     3301 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     3361 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     3421 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     3481 aacgcgctga gggccgccac tccaccggcg gcatggacga gctgtacaag taaccaatta
     3541 ttgaagggga gcgggaaacc gctccccttt ttttgtttct ggtctcccta cta
//
