LOCUS       sequence-65427-e        2064 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 2064)
  AUTHORS   Huang L, Jin J, Deighan P, Kiner E, McReynolds L, Lieberman J
  TITLE     Efficient and specific gene knockdown by small interfering RNAs 
            produced in bacteria.
  JOURNAL   Nat Biotechnol. 2013 Apr;31(4):350-6. doi: 10.1038/nbt.2537. Epub 
            2013 Mar 10.
  PUBMED    23475073
REFERENCE   2  (bases 1 to 2064)
  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.2537"; 
            journalName: "Nat Biotechnol"; date: "2013-04"; volume: "31"; issue:
            "4"; pages: "350-6"
COMMENT     Sequence Label: pGEX-4T-1-p19-T7-EGFPFL
FEATURES             Location/Qualifiers
     source          1..2064
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             523..540
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     547..566
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        547..565
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(573..575,576..578,579..1292)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(573..575,576..578,579..1289)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1337..2050,2051..2053,2054..2056))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1340..2050,2051..2053,2054..2056))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
ORIGIN
        1 ggatccatgg aacgagctat acaaggaaac gacgctaggg aacaagctaa cagtgaacgt
       61 tgggatggag gatcaggagg taccacttct cccttcaaac ttcctgacga aagtccgagt
      121 tggactgagt ggcggctaca taacgatgag acgaattcga atcaagataa tccccttggt
      181 ttcaaggaaa gctggggttt cgggaaagtt gtatttaaga gatatctcag atacgacagg
      241 acggaagctt cactgcacag agtccttgga tcttggacgg gagattcggt taactatgca
      301 gcatctcgat ttttcggttt cgaccagatc ggatgtacct atagtattcg gtttcgagga
      361 gttagtatca ccgtttctgg agggtcgcga actcttcagc atctctgtga gatggcaatt
      421 cggtctaagc aagaactgct acagcttgcc ccaatcgaag tggaaagtaa tgtatcaaga
      481 ggatgccctg aaggtactca aaccttcgaa aaagaaagcg agcaccacca ccaccaccac
      541 tgaatctaat acgactcact ataggggagc tcatggtgag caagggcgag gagctgttca
      601 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
      661 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
      721 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
      781 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
      841 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
      901 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
      961 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1021 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     1081 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1141 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     1201 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1261 tcactctcgg catggacgag ctgtacaagt agctcgagtc tagagggccc gtttaaaccc
     1321 gctgatcagc gtcgacctac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg
     1381 tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg
     1441 actgggtgct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt
     1501 tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga
     1561 agttcacctt gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt
     1621 agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca
     1681 gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt
     1741 tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact
     1801 tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg
     1861 tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact
     1921 tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt
     1981 ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct
     2041 cgcccttgct caccatgcgg ccgc
//
