LOCUS       sequence-313036-        1659 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Constitutive expression of 3xFLAG-NLS-DsDed-ZF10-intN-VP64 under the
            EF1-alpha promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1659)
  AUTHORS   Muldoon JJ, Kandula V, Hong M, Donahue PS, Boucher JD, Bagheri N, 
            Leonard JN
  TITLE     Model-guided design of mammalian genetic programs.
  JOURNAL   Sci Adv. 2021 Feb 19;7(8). pii: 7/8/eabe9375. doi: 
            10.1126/sciadv.abe9375. Print 2021 Feb.
  PUBMED    33608279
REFERENCE   2  (bases 1 to 1659)
  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: "Sci Adv."; 
            date: "2021-02-19"; volume: "7(8). pii: 7/8/eabe9375. doi"; pages: "
            10.1126/sciadv.abe9375. Print 2021 Feb"
COMMENT     Sequence Label: pJM592 EF1-alpha 3xFLAG-NLS-DsDed-ZF10-intN-VP64 
            in...
FEATURES             Location/Qualifiers
     source          1..1659
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             4..69
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             76..96
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             139..813
                     /codon_start=1
                     /product="noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2008)"
                     /label=DsRed-Express2
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWEKVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     complement(319..339)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     731..754
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             1507..1656
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
ORIGIN
        1 atggattata aggatcacga tggcgactac aaggaccacg atattgacta caaagacgat
       61 gacgataaaa tggctcctaa gaaaaagcgc aaagtcggta tccatggcgt tcccggatca
      121 ggctccggaa gtggctccat ggatagcact gagaacgtca tcaagccctt catgcgcttc
      181 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
      241 ggcaagccct acgagggcac ccagaccgcc aagctccagg tgaccaaggg cggccccctg
      301 cccttcgcct gggacatcct gtccccccag ttccagtacg gctccaaggt gtacgtgaag
      361 caccccgccg acatccccga ctacaagaag ctgtccttcc ccgagggctt caagtgggag
      421 aaggtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctccag
      481 gacggcacct tcatctacca cgtgaagttc atcggcgtga acttcccctc cgacggcccc
      541 gtaatgcaga agaaaactct gggctgggag ccctccaccg agcgcctgta cccccgcgac
      601 ggcgtgctga agggcgagat ccacaaggcg ctgaagctga agggcggcgg ccactacctg
      661 gtggagttca agtcaatcta catggccaag aagcccgtga agctgcccgg ctactactac
      721 gtggactcca agctggacat cacctcccac aacgaggact acaccgtggt ggagcagtac
      781 gagcgcgccg aggcccgcca ccacctgttc cagtctggtt ctggttctgg ttcaggatcc
      841 ggtaccggtg gagctagacc cggggagcgc cccttccagt gtcgcatttg catgcggaac
      901 ttttcgacta atcaaaaatt ggaagttcat acccgtactc ataccggtga aaaaccgttt
      961 cagtgtcgga tctgtatgcg aaatttctcc gttagacata atttgcaaag acatctacgt
     1021 acgcacaccg gcgagaagcc attccaatgc cgaatatgca tgcgcaactt cagtcaacat
     1081 ccaaatttga ctagacacct aaaaacccac ctgagaggat ctacttctgg tggtggcggg
     1141 tctggaggag gaggtactgg tggaggtggc actggtacca gaagcggata ctgcctggac
     1201 ctgaagaccc aggtgcagac ccctcagggc atgaaggaga tcagcaacat ccaggtgggc
     1261 gacctggtgc tgagcaacac cggctacaac gaggtgctga acgtgttccc caagagcaag
     1321 aagaagagct acaagatcac cctggaggac ggcaaggaga tcatctgcag cgaggagcac
     1381 ctgttcccca cccagaccgg cgagatgaac atcagcggcg gcctgaagga gggcatgtgc
     1441 ctgtacgtga aggagggagg cggaagcgga ggcggtggat caggctccgg aagtggctcc
     1501 gaattcgacg cccttgatga ctttgacttg gacatgctgg ggtcagacgc actcgatgat
     1561 tttgaccttg atatgctggg aagcgatgca cttgatgact ttgacttgga catgctcggc
     1621 tccgacgctc ttgatgattt cgatcttgac atgctctag
//
