LOCUS       sequence-478606-         993 bp    DNA     linear   UNA 14-JUL-2025
DEFINITION  Expresses eGFP fluorophore with fused p2c targeting ligand for 
            expression in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 993)
  AUTHORS   Chaverra-Rodriguez D, Macias VM, Hughes GL, Pujhari S, Suzuki Y, 
            Peterson DR, Kim D, McKeand S, Rasgon JL
  TITLE     Targeted delivery of CRISPR-Cas9 ribonucleoprotein into arthropod 
            ovaries for heritable germline gene editing.
  JOURNAL   Nat Commun. 2018 Aug 1;9(1):3008. doi: 10.1038/s41467-018-05425-9.
  PUBMED    30068905
REFERENCE   2  (bases 1 to 993)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2018-08-1"; volume: "9(1):3008. doi"; pages: " 
            10.1038/s41467-018-05425-9"
COMMENT     Sequence Label: pET28(a)-p2c-eGFP
FEATURES             Location/Qualifiers
     source          1..993
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1..18
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             28..45
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             49..81
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(274..276,277..279,280..987)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKGD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(319..340)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     927..948
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 catcatcatc atcatcacag cagcggcctg gtgccgcgcg gcagccatat ggctagcatg
       61 actggtggac agcaaatggg tcgcggatcc aatctgcagc agcagcgcca gcacggcaag
      121 aacggcaacc aggactacca ggatcagagc aacgaacaga ggaagaacca gaggaccagc
      181 agcgaggagg actacagcga ggaggttaag aacgagctca ggcgcgccgg aggaggcggc
      241 agcggtggtg gaggaagcgg tggaggcggg agcatggtga gcaagggcga ggagctgttc
      301 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      361 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
      421 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
      481 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
      541 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg gcgacggcaa ctacaagacc
      601 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
      661 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
      721 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
      781 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
      841 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
      901 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
      961 atcactctcg gcatggacga gctgtacgga ttc
//
