LOCUS       sequence-138402-         885 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  EGFP coding region followed by enhanced synthetic inhibitory 
            BIC/miR-155 RNA (eSIBR) backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 885)
  AUTHORS   Fowler DK, Williams C, Gerritsen AT, Washbourne P
  TITLE     Improved knockdown from artificial microRNAs in an enhanced miR-155 
            backbone: a designer's guide to potent multi-target RNAi.
  JOURNAL   Nucleic Acids Res. 2015 Nov 17. pii: gkv1246.
  PUBMED    26582923
REFERENCE   2  (bases 1 to 885)
  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: "Nucleic 
            Acids Res. 2015 Nov 17. pii: gkv1246."
COMMENT     Sequence Label: GFP-eSIBR
FEATURES             Location/Qualifiers
     source          1..885
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     36..55
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        36..54
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      68..77
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(74..76,77..79,80..793)
                     /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(74..76,77..79,80..790)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(119..140)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(380..399)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     727..748
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
       61 caagcttgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
      121 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
      181 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
      241 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
      301 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
      361 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
      421 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
      481 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
      541 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
      601 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
      661 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
      721 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
      781 gctgtacaag taaggatcca ctagtaacgg ccgccagtgt gctggaattc aaagtcgact
      841 ggaggcttgc tttgggctgt atgctgttgt cttcaagatc tggaa
//
