LOCUS       sequence-463248-        3039 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Lentiviral expression of ComMAND closed-loop circuit regulating 
            EGFP-FMRP (therapeutically relevant gene).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3039)
  AUTHORS   Love KS, Johnstone CP, Peterman EL, Gaglione S, Birnbaum ME, 
            Galloway KE
  TITLE     Model-guided design of microRNA-based gene circuits supports precise
            dosage of transgenic cargoes into diverse primary cells.
  JOURNAL   Cell Syst. 2025 Apr 22:101269. doi: 10.1016/j.cels.2025.101269.
  PUBMED    40300600
REFERENCE   2  (bases 1 to 3039)
  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: "Cell Syst. 
            2025 Apr 22:101269. doi: 10.1016/j.cels.2025.101269."
COMMENT     Sequence Label: 
            pLentiX1-[TRE3G-EGFP(miRE.FF4)-FMRP-TS.FF4x1-bGH]-EFS-rtTA-P2A-mRuby
            2-WPRE
FEATURES             Location/Qualifiers
     source          1..3039
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(46..67)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(307..326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     ncRNA           527..541
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           598..710
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     primer_bind     1071..1092
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1177..1200
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      121 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      301 ttcaaggacg acggcaacta caagacccgc gccgaggtaa gtgtgctcgc ttcggcagca
      361 catatactat gttgaatgag gcttcagtac tttacagaat cgttgcctgc acatcttgga
      421 aacacttgct gggattactt cttcagggac ttcttaaccc aacagaaggc tcgagaaggt
      481 atattgctgt tgacagtgag cgccgcttga agtctttaat taaatagtga agccacagat
      541 gtatttaatt aaagacttca agcggtgcct actgcctcgg acttcaaggg gctagaattc
      601 aaggggctac tttaggagca attatcttgt ttactaaaac tgaatacctt gctatctctt
      661 tgatacattt ttacaaagct gaattaaaat ggtataaatt aaatcacttt tttcaattgt
      721 actaacttcg agtcttcttt ttttttttca caggtgaagt tcgagggcga caccctggtg
      781 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
      841 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
      901 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
      961 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1021 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1081 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtccgga
     1141 ctcagatctc gagctcaagc ttcgaattcg atggaggact acaaggacga cgatgacaag
     1201 gagctggtgg tggaagtgcg gggctccaat ggcgctttct acaaggcatt tgtaaaagat
     1261 gtccatgaag attctataac agttgctttt gaaaacaact ggcaaccaga gagacagatt
     1321 ccattccatg atgtgagatt cccaccacct gtaggttata ataaagatat aaacgaaagt
     1381 gatgaagttg aggtttattc cagagcaaat gaaaaagagc cttgctgttg gtggttagct
     1441 aaagtgagga tgataaaggg tgagttttat gtgatagaat atgcagcatg tgatgctacg
     1501 tataatgaaa ttgtcacaat tgagcgtcta cgatctgtta atcccaacaa acctgctaca
     1561 aaagatactt tccataagat caagctggag gtgccagaag atttacgaca aatgtgtgcc
     1621 aaagaatcag cacataagga ttttaaaaag gcagttggtg ccttctctgt aacttatgat
     1681 ccagaaaatt atcagctggt aattttgtcc atcaatgaag tcacctcaaa gcgagcccac
     1741 atgttgattg acatgcactt tcgaagtctg cgcaccaagt tgtctcttat actgagaaat
     1801 gaagaagcca gtaaacaact ggagagttca aggcagcttg cctcaagatt tcatgaacag
     1861 tttatcgtac gagaagatct gatgggttta gctattggta ctcatggtgc taatattcag
     1921 caagctagaa aagtgcctgg cgtcactgct attgatttag atgaggatac ctgcacattt
     1981 catatttatg gagaggatca agatgcagtg aaaaaggcta gaagctttct ggaatttgct
     2041 gaagatgtca tacaggttcc acgaaactta gtaggcaaag taataggaaa aaatggaaag
     2101 ctgattcaag agatcgtgga caagtcagga gttgtgaggg tgaggattga ggctgaaaat
     2161 gagaaaagtg tcccacaaga agaggaaatt atgccaccaa gttccctacc ttccaataat
     2221 tcaagggttg gacctaactc ctctgaagaa aagaaacatt tagatacaaa ggaaaacacc
     2281 catttttctc aacctaacag tacaaaagtc cagagggtgt tagtggtttc atcaattgta
     2341 gcagggggac cccagaaacc tgaacccaag gcttggcagg gtatggtacc atttgttttt
     2401 gtgggaacaa aagacagcat cgctaatgcc actgttcttt tggattatca cctgaactat
     2461 ttaaaggaag tagaccagtt gcgtttggag agattacaaa ttgatgagca gttgcgacaa
     2521 attggagcta gttctagacc accaccaaat cgtacagata aggaaaaagg ctatgtgact
     2581 gatgatggtc aaggaatggg tcgaggtagt agaccttaca gaaatagggg gcacggcaga
     2641 cgcggtcctg gatatacttc aggaactaat tctgaagcat caaatgcttc tgaaacagaa
     2701 tctgaccaca gagacgaact cagtgattgg tcattagctc caacagagga agagagggag
     2761 agcttcctgc gcagaggaga cggacggcgg cgtggaggag gaggaagagg acaaggagga
     2821 agaggaagag gaggaggctt caaaggaaac gacgatcatt cccgaacaga taatcgtcca
     2881 cgtaatccaa gagaggctaa aggaagaaca gctgatggat ccctgcagag tgcctccagt
     2941 gaagggagcc ggctgcgcac gggtaaagat cgtaaccaga agaaggaaaa gccagacagc
     3001 gtagatgggc tgcaaccgct ggtgaatgga gtaccctaa
//
