LOCUS       sequence-43209-e        3531 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3531)
  AUTHORS   Zhang F, Cong L, Lodato S, Kosuri S, Church GM, Arlotta P
  TITLE     Efficient construction of sequence-specific TAL effectors for 
            modulating mammalian transcription.
  JOURNAL   Nat Biotechnol. 2011 Feb;29(2):149-53. doi: 10.1038/nbt.1775
  PUBMED    21248753
REFERENCE   2  (bases 1 to 3531)
  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: "10.1038/nbt"; 
            journalName: "Nat Biotechnol"; date: "2011-02"; volume: "29"; issue:
            "2"; pages: "149-53"
COMMENT     Sequence Label: pLenti-EF1a-KLF4
FEATURES             Location/Qualifiers
     source          1..3531
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             2545..2565
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2590..2739
                     /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             2761..2814
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(2815..2817,2818..3531)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2815..2817,2818..3528)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2857..2878)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3118..3137)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3465..3486
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atgtcgcgga cccggctccc ttccccaccc gcacccagcc cagcgttttc ggccgactcg
       61 ttctcagacc tgcttaggca gttcgacccc tcactgttta acacatcgtt gttcgactcc
      121 cttcctccgt ttggggcgca ccatacggag gcggccaccg gggagtggga tgaggtgcag
      181 tcgggattga gagctgcgga tgcaccaccc ccaaccatgc gggtggccgt caccgctgcc
      241 cgaccgccga gggcgaagcc cgcaccaagg cggagggcag cgcaaccgtc cgacgcaagc
      301 cccgcagcgc aagtagattt gagaactttg ggatattcac agcagcagca ggaaaagatc
      361 aagcccaaag tgaggtcgac agtcgcgcag catcacgaag cgctggtggg tcatgggttt
      421 acacatgccc acatcgtagc cttgtcgcag caccctgcag cccttggcac ggtcgccgtc
      481 aagtaccagg acatgattgc ggcgttgccg gaagccacac atgaggcgat cgtcggtgtg
      541 gggaaacagt ggagcggagc ccgagcgctt gaggccctgt tgacggtcgc gggagagctg
      601 agagggcctc cccttcagct ggacacgggc cagttgctga agatcgcgaa gcggggagga
      661 gtcacggcgg tcgaggcggt gcacgcgtgg cgcaatgcgc tcacgggagc acccctcaac
      721 ctgaccccag agcaggtcgt ggccattgcc tcgaatggag ggggcaaaca ggcgttggaa
      781 accgtacaac gattgctgcc ggtgctgtgc caagcgcacg gactcacccc agagcaggtc
      841 gtggcgatcg caagccacga cggaggaaag caagccttgg aaacagtaca gaggctgttg
      901 cctgtgctgt gccaagcgca cggcctcacc ccagagcagg tcgtggccat tgcctcgaat
      961 ggagggggca aacaggcgtt ggaaaccgta caacgattgc tgccggtgct gtgccaagcg
     1021 cacggattaa ccccagagca ggtcgtggcc attgcctcga atggaggggg caaacaggcg
     1081 ttggaaaccg tacaacgatt gctgccggtg ctgtgccaag cgcacggctt aaccccagag
     1141 caggtcgtgg caatcgcctc caacattggc gggaaacagg cactcgagac tgtccagcgc
     1201 ctgcttcccg tgctgtgcca agcgcacgga cttaccccag agcaggtcgt ggcgatcgca
     1261 agccacgacg gaggaaagca agccttggaa acagtacaga ggctgttgcc tgtgctgtgc
     1321 caagcgcacg gccttacccc agagcaggtc gtggccattg cctcgaatgg agggggcaaa
     1381 caggcgttgg aaaccgtaca acgattgctg ccggtgctgt gccaagcgca cggactaacc
     1441 ccagagcagg tcgtggccat tgcctcgaat ggagggggca aacaggcgtt ggaaaccgta
     1501 caacgattgc tgccggtgct gtgccaagcg cacgggctca ccccagagca ggtcgtggca
     1561 atcgcctcca acattggcgg gaaacaggca ctcgagactg tccagcgcct gcttcccgtg
     1621 ctgtgccaag cgcacgggct aaccccagag caggtcgtgg ccattgcctc gaatggaggg
     1681 ggcaaacagg cgttggaaac cgtacaacga ttgctgccgg tgctgtgcca agcgcacggc
     1741 ctaaccccag agcaggtcgt ggcaatcgcc tccaacattg gcgggaaaca ggcactcgag
     1801 actgtccagc gcctgcttcc cgtgctgtgc caagcgcacg ggttaacccc agagcaggtc
     1861 gtggcaatcg cctccaacat tggcgggaaa caggcactcg agactgtcca gcgcctgctt
     1921 cccgtgctgt gccaagcgca cggactcacg cctgagcagg tagtggctat tgcatcccat
     1981 gacgggggca gacccgcact ggagtcaatc gtggcccagc tttcgaggcc ggaccccgcg
     2041 ctggccgcac tcactaatga tcatcttgta gcgctggcct gcctcggcgg acgacccgcc
     2101 ttggatgcgg tgaagaaggg gctcccgcac gcgcctgcat tgattaagcg gaccaacaga
     2161 aggattcccg agaggacatc acatcgagtg gcagatcacg cgcaagtggt ccgcgtgctc
     2221 ggattcttcc agtgtcactc ccaccccgca caagcgttcg atgacgccat gactcaattt
     2281 ggtatgtcga gacacggact gctgcagctc tttcgtagag tcggtgtcac agaactcgag
     2341 gcccgctcgg gcacactgcc tcccgcctcc cagcggtggg acaggattct ccaagcgagc
     2401 ggtatgaaac gcgcgaagcc ttcacctacg tcaactcaga cacctgacca ggcgagcctt
     2461 catgcgttcg cagactcgct ggagagggat ttggacgcgc cctcgcccat gcatgaaggg
     2521 gaccaaactc gcgcgtcagc tagccccaag aagaagagaa aggtggaggc cagcggttcc
     2581 ggacgggctg acgcattgga cgattttgat ctggatatgc tgggaagtga cgccctcgat
     2641 gattttgacc ttgacatgct tggttcggat gcccttgatg actttgacct cgacatgctc
     2701 ggcagtgacg cccttgatga tttcgacctg gacatgctga ttaactctag aggcagtgga
     2761 gagggcagag gaagtctgct aacatgcggt gacgtcgagg agaatcctgg cccagtgagc
     2821 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     2881 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     2941 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     3001 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     3061 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     3121 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     3181 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     3241 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     3301 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     3361 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     3421 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     3481 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta a
//
