LOCUS       sequence-401595-        3871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes mCherry, which can be expressed with the activation of a 
            synthetic promoter by Cas enzymes. A cloning site is provided to 
            construct the synthetic Cas-targeted promoter upstream of mCherry..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3871)
  AUTHORS   Kang H, Fitch JC, Varghese RP, Thorne CA, Cusanovich DA
  TITLE     Optimization of a Cas12a-Driven Synthetic Gene Regulatory Network 
            System.
  JOURNAL   ACS Synth Biol. 2025 May 16;14(5):1732-1744. doi: 
            10.1021/acssynbio.5c00084. Epub 2025 May 2.
  PUBMED    40316310
REFERENCE   2  (bases 1 to 3871)
  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: 
            "10.1021/acssynbio.5c00084"; journalName: "ACS Synth Biol"; date: 
            "2025-05-16- 16"; volume: "14"; issue: "5"; pages: "1732-1744"
COMMENT     Sequence Label: Px-mCherry-pA
FEATURES             Location/Qualifiers
     source          1..3871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(177..968,969..1037))
                     /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     800..819
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1038..1142)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1169..1624
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1256..1275)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1466..1487
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1755..1803
                     /note="synthetic polyadenylation signal"
     misc_feature    1817..1908
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     1857..1876
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     primer_bind     1922..1938
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1922..1938
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2008..2718
                     /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(2153..2171)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2421..2440
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2744..2865
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2755..2774)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2809..2828
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3118..3135)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(3289..3871,1..6))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3369..3388)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
       61 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
      121 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
      181 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
      241 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
      301 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
      361 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
      421 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
      481 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
      541 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
      601 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
      661 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
      721 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
      781 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
      841 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
      901 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
      961 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     1021 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     1081 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     1141 cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg
     1201 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     1261 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     1321 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     1381 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     1441 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     1501 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     1561 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     1621 atttgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     1681 tcgctattac gccagcccaa gctaccatga taagtaagta atattaaggt acgggaggta
     1741 cttggagcgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt
     1801 gtgaatcgat agtactaaca tacgctctcc atcaaaacaa aacgaaacaa aacaaactag
     1861 caaaataggc tgtccccagt gcaagtgcag gtgccagaac atttctctat cgataggatc
     1921 ccaggaaaca gctatgacga gctctctagc ggggggctat aaaagggggt gggggcgttc
     1981 gtcctgctat ctagcgtcgc gttgaccatg gtgagcaagg gcgaggagga taacatggcc
     2041 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     2101 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     2161 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     2221 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     2281 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     2341 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     2401 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     2461 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     2521 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     2581 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     2641 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     2701 gacgagctgt acaagtaagg ccggccgctt cgagcagaca tgataagata cattgatgag
     2761 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     2821 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     2881 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     2941 ctctacaaat gtggtaaaat cgataaggat cgagtatgtc gaccgatgcc cttgagagcc
     3001 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     3061 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc
     3121 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3181 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     3241 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     3301 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3361 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3421 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3481 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3541 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3601 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3661 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3721 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3781 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3841 caagcagcag attacgcgca gaaaaaaagg a
//
