LOCUS       sequence-463242-        2622 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Lentiviral expression of ComMAND base gene regulating EGFP-FMRP 
            (therapeutically relevant gene).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2622)
  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 2622)
  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-FMRP-bGH]-EFS-rtTA-P2A-mRuby2-WPRE
FEATURES             Location/Qualifiers
     source          1..2622
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     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"
     primer_bind     654..675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             760..783
                     /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 gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
      481 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtcc
      721 ggactcagat ctcgagctca agcttcgaat tcgatggagg actacaagga cgacgatgac
      781 aaggagctgg tggtggaagt gcggggctcc aatggcgctt tctacaaggc atttgtaaaa
      841 gatgtccatg aagattctat aacagttgct tttgaaaaca actggcaacc agagagacag
      901 attccattcc atgatgtgag attcccacca cctgtaggtt ataataaaga tataaacgaa
      961 agtgatgaag ttgaggttta ttccagagca aatgaaaaag agccttgctg ttggtggtta
     1021 gctaaagtga ggatgataaa gggtgagttt tatgtgatag aatatgcagc atgtgatgct
     1081 acgtataatg aaattgtcac aattgagcgt ctacgatctg ttaatcccaa caaacctgct
     1141 acaaaagata ctttccataa gatcaagctg gaggtgccag aagatttacg acaaatgtgt
     1201 gccaaagaat cagcacataa ggattttaaa aaggcagttg gtgccttctc tgtaacttat
     1261 gatccagaaa attatcagct ggtaattttg tccatcaatg aagtcacctc aaagcgagcc
     1321 cacatgttga ttgacatgca ctttcgaagt ctgcgcacca agttgtctct tatactgaga
     1381 aatgaagaag ccagtaaaca actggagagt tcaaggcagc ttgcctcaag atttcatgaa
     1441 cagtttatcg tacgagaaga tctgatgggt ttagctattg gtactcatgg tgctaatatt
     1501 cagcaagcta gaaaagtgcc tggcgtcact gctattgatt tagatgagga tacctgcaca
     1561 tttcatattt atggagagga tcaagatgca gtgaaaaagg ctagaagctt tctggaattt
     1621 gctgaagatg tcatacaggt tccacgaaac ttagtaggca aagtaatagg aaaaaatgga
     1681 aagctgattc aagagatcgt ggacaagtca ggagttgtga gggtgaggat tgaggctgaa
     1741 aatgagaaaa gtgtcccaca agaagaggaa attatgccac caagttccct accttccaat
     1801 aattcaaggg ttggacctaa ctcctctgaa gaaaagaaac atttagatac aaaggaaaac
     1861 acccattttt ctcaacctaa cagtacaaaa gtccagaggg tgttagtggt ttcatcaatt
     1921 gtagcagggg gaccccagaa acctgaaccc aaggcttggc agggtatggt accatttgtt
     1981 tttgtgggaa caaaagacag catcgctaat gccactgttc ttttggatta tcacctgaac
     2041 tatttaaagg aagtagacca gttgcgtttg gagagattac aaattgatga gcagttgcga
     2101 caaattggag ctagttctag accaccacca aatcgtacag ataaggaaaa aggctatgtg
     2161 actgatgatg gtcaaggaat gggtcgaggt agtagacctt acagaaatag ggggcacggc
     2221 agacgcggtc ctggatatac ttcaggaact aattctgaag catcaaatgc ttctgaaaca
     2281 gaatctgacc acagagacga actcagtgat tggtcattag ctccaacaga ggaagagagg
     2341 gagagcttcc tgcgcagagg agacggacgg cggcgtggag gaggaggaag aggacaagga
     2401 ggaagaggaa gaggaggagg cttcaaagga aacgacgatc attcccgaac agataatcgt
     2461 ccacgtaatc caagagaggc taaaggaaga acagctgatg gatccctgca gagtgcctcc
     2521 agtgaaggga gccggctgcg cacgggtaaa gatcgtaacc agaagaagga aaagccagac
     2581 agcgtagatg ggctgcaacc gctggtgaat ggagtaccct aa
//
