LOCUS       sequence-313032-        1554 bp    DNA     linear   UNA 15-APR-2025
DEFINITION  Constitutive expression of 3xFLAG-NLS-VP64-intC-ZF1-DsDed-NLS-HA 
            under the EF1-alpha promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1554)
  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 1554)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pJM588 EF1-alpha 
            3xFLAG-NLS-VP64-intC-ZF1-DsDed-NLS...
FEATURES             Location/Qualifiers
     source          1..1554
                     /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             112..261
                     /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"
     CDS             814..1488
                     /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(994..1014)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1406..1429
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             1498..1518
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1525..1551
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 atggactata aagaccatga cggtgactac aaggatcacg acatagatta taaagacgat
       61 gacgacaaga tggcacctaa gaagaagcgc aaagttggct ctggcgaatt cgacgccctt
      121 gatgactttg acttggacat gctggggtca gacgcactcg atgattttga ccttgatatg
      181 ctgggaagcg atgcacttga tgactttgac ttggacatgc tcggctccga cgctcttgat
      241 gatttcgatc ttgacatgct cggatccggt ggtggcgggt ctggtggagg aggtagtggt
      301 ggaggtggct ctgccaccat gatgctgaag aagatcctga agatcgagga gctggacgag
      361 agagagctga tcgacatcga ggtgagcggc aaccacctgt tctacgccaa cgacatcctg
      421 acccacaaca gcaacagggg tggtggcggg tctggtggag gaggtagtgg tggaggtggc
      481 tctggtaccg ctagacccgg ggagcgcccc ttccagtgtc gcatttgcat gcggaacttt
      541 tcgcgccagg acaggcttga caggcatacc cgtactcata ccggtgaaaa accgtttcag
      601 tgtcggatct gtatgcgaaa tttctcccag aaggagcact tggcggggca tctacgtacg
      661 cacaccggcg agaagccatt ccaatgccga atatgcatgc gcaacttcag tcgccgcgac
      721 aacctgaacc ggcacctaaa aacccacctg agagaattcg gctctggcgg aggcggaagc
      781 ggaggcggtg gatcaggctc cggaagtggc tccatggata gcactgagaa cgtcatcaag
      841 cccttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
      901 gagggcgagg gcgagggcaa gccctacgag ggcacccaga ccgccaagct ccaggtgacc
      961 aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcca gtacggctcc
     1021 aaggtgtacg tgaagcaccc cgccgacatc cccgactaca agaagctgtc cttccccgag
     1081 ggcttcaagt gggagaaggt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
     1141 gactcctccc tccaggacgg caccttcatc taccacgtga agttcatcgg cgtgaacttc
     1201 ccctccgacg gccccgtaat gcagaagaaa actctgggct gggagccctc caccgagcgc
     1261 ctgtaccccc gcgacggcgt gctgaagggc gagatccaca aggcgctgaa gctgaagggc
     1321 ggcggccact acctggtgga gttcaagtca atctacatgg ccaagaagcc cgtgaagctg
     1381 cccggctact actacgtgga ctccaagctg gacatcacct cccacaacga ggactacacc
     1441 gtggtggagc agtacgagcg cgccgaggcc cgccaccacc tgttccagtc tggttctcct
     1501 aagaagaagc gcaaagttgg catgtaccca tacgacgtac cagattacgc ttag
//
