LOCUS       sequence-482809-        4431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low copy mRFP1 reporter for monitoring metabolite-responsive CRISPRa
            activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4431)
  AUTHORS   Stohr AM, Hansen H, Richards B, Park H, Goncalves AG, Agrawal A, 
            Blenner M, Chen W
  TITLE     Metabolite-responsive scaffold RNAs for dynamic CRISPR 
            transcriptional regulation.
  JOURNAL   Nucleic Acids Res. 2025 Nov 13;53(21):gkaf1290. doi: 
            10.1093/nar/gkaf1290.
  PUBMED    41277686
REFERENCE   2  (bases 1 to 4431)
  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."; date: "2025-11-13"; volume: "53(21):gkaf1290. doi"; 
            pages: " 10.1093/nar/gkaf1290"
COMMENT     Sequence Label: pSC101-MP-mRFP1
FEATURES             Location/Qualifiers
     source          1..4431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..951)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(1713..1807)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(1837..2628,2629..2697))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2460..2479
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2698..2802)
                     /gene="bla"
                     /label=AmpR promoter
     RBS             3059..3070
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3077..3754
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      3787..3858
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3874..3901
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag
       61 ccatgagaac gaaccattga gatcatactt actttgcatg tcactcaaaa attttgcctc
      121 aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt
      181 atgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg
      241 gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa
      301 cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa
      361 atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt
      421 ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt
      481 tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc
      541 aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg
      601 caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt
      661 ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat
      721 cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat
      781 ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat
      841 cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc
      901 atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa
      961 ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat gataattact
     1021 agtccttttc ccgggtgatc tgggtatctg taaattctgc tagacctttg ctggaaaact
     1081 tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta
     1141 tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga
     1201 tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc
     1261 gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac
     1321 cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga
     1381 gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg
     1441 gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag
     1501 aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct
     1561 gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta
     1621 tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca
     1681 ggcttacccg tcttactgtc cctagtgctt ggattctcac caataaaaaa cgcccggcgg
     1741 caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac
     1801 aggagtccaa gcgagctcgt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     1861 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     1921 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     1981 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2041 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2101 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     2161 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     2221 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     2281 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     2341 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     2401 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     2461 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     2521 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     2581 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     2641 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     2701 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     2761 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     2821 gccacctgtg gcaattccga cgtcagcatt tgcgatcatt cacgcagcgc ttattcagtt
     2881 gctcactgcg atgtcataat catcgctacg agctgtgaaa gatgcataaa gctcgtacga
     2941 cgcgttcgct cgtctcctca cttctcctaa ctctcacacg tggctgcacc ggcccccccc
     3001 gctgccgcgg gccgttgaca gtttcggggc cgaaagggat tgtacgagcg aattcattaa
     3061 agaggagaaa ggtaccatgg cgagtagcga agacgttatc aaagagttca tgcgtttcaa
     3121 agttcgtatg gaaggttccg ttaacggtca cgagttcgaa atcgaaggtg aaggtgaagg
     3181 tcgtccgtac gaaggtaccc agaccgctaa actgaaagtt accaaaggtg gtccgctgcc
     3241 gttcgcttgg gacatcctgt ccccgcagtt ccagtacggt tccaaagctt acgttaaaca
     3301 cccggctgac atcccggact acctgaaact gtccttcccg gaaggtttca aatgggaacg
     3361 tgttatgaac ttcgaagacg gtggtgttgt taccgttacc caggactcct ccctgcaaga
     3421 cggtgagttc atctacaaag ttaaactgcg tggtaccaac ttcccgtccg acggtccggt
     3481 tatgcagaaa aaaaccatgg gttgggaagc ttccaccgaa cgtatgtacc cggaagacgg
     3541 tgctctgaaa ggtgaaatca aaatgcgtct gaaactgaaa gacggtggtc actacgacgc
     3601 tgaagttaaa accacctaca tggctaaaaa accggttcag ctgccgggtg cttacaaaac
     3661 cgacatcaaa ctggacatca cctcccacaa cgaagactac accatcgttg aacagtacga
     3721 acgtgctgaa ggtcgtcact ccaccggtgc ttaaggatcc aaactcgagt aaggatctcc
     3781 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     3841 ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc
     3901 gtttatacct agggtacggg ttttgctgcc cgcaaacggg ctgttctggt gttgctagtt
     3961 tgttatcaga atcgcagatc cggcttcagc cggtttgccg gctgaaagcg ctatttcttc
     4021 cagaattgcc atgatttttt ccccacggga ggcgtcactg gctcccgtgt tgtcggcagc
     4081 tttgattcga taagcagcat cgcctgtttc aggctgtcta tgtgtgactg ttgagctgta
     4141 acaagttgtc tcaggtgttc aatttcatgt tctagttgct ttgttttact ggtttcacct
     4201 gttctattag gtgttacatg ctgttcatct gttacattgt cgatctgttc atggtgaaca
     4261 gctttgaatg caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt tacaccgttt
     4321 tcatctgtgc atatggacag ttttcccttt gatatgtaac ggtgaacagt tgttctactt
     4381 ttgtttgtta gtcttgatgc ttcactgata gatacaagag ccataagaac c
//
