LOCUS       sequence-222687-        2556 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2556)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 2556)
  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"
COMMENT     Sequence Label: CHL1 wt-eGFP
FEATURES             Location/Qualifiers
     source          1..2556
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1780..1800
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             join(1807..1809,1810..1812,1813..2523)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1807..1809,1810..1812,1813..2523)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1852..1873)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2113..2132)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2460..2481
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2530..2553
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
ORIGIN
        1 atgtctcttc ctgaaactaa atctgatgat atccttcttg atgcttggga cttccaaggc
       61 cgtcccgccg atcgctcaaa aaccggcggc tgggccagcg ccgccatgat tctttgtatt
      121 gaggccgtgg agaggctgac gacgttaggt atcggagtta atctggtgac gtatttgacg
      181 ggaactatgc atttaggcaa tgcaactgcg gctaacaccg ttaccaattt cctcggaact
      241 tctttcatgc tctgtctcct cggtggcttc atcgccgata cctttctcgg caggtaccta
      301 acgattgcta tattcgccgc aatccaagcc acgggtgttt caatcttaac tctatcaaca
      361 atcataccgg gacttcgacc accaagatgc aatccaacaa cgtcgtctca ctgcgaacaa
      421 gcaagtggaa tacaactgac ggtcctatac ttagccttat acctcaccgc tctaggaacg
      481 ggaggcgtga aggctagtgt ctcgggtttc gggtcggacc aattcgatga gaccgaacca
      541 aaagaacgat cgaaaatgac atatttcttc aaccgtttct tcttttgtat caacgttggc
      601 tctcttttag ctgtgacggt ccttgtctac gtacaagacg atgttggacg caaatggggc
      661 tatggaattt gcgcgtttgc gatcgtgctt gcactcagcg ttttcttggc cggaacaaac
      721 cgctaccgtt tcaagaagtt gatcggtagc ccgatgacgc aggttgctgc ggttatcgtg
      781 gcggcgtgga ggaataggaa gctcgagctg ccggcagatc cgtcctatct ctacgatgtg
      841 gatgatatta ttgcggcgga aggttcgatg aagggtaaac aaaagctgcc acacactgaa
      901 caattccgtt cattagataa ggcagcaata agggatcagg aagcgggagt tacctcgaat
      961 gtattcaaca agtggacact ctcaacacta acagatgttg aggaagtgaa acaaatcgtg
     1021 cgaatgttac caatttgggc aacatgcatc ctcttctgga ccgtccacgc tcaattaacg
     1081 acattatcag tcgcacaatc cgagacattg gaccgttcca tcgggagctt cgagatccct
     1141 ccagcatcga tggcagtctt ctacgtcggt ggcctcctcc taaccaccgc cgtctatgac
     1201 cgcgtcgcca ttcgtctatg caaaaagcta ttcaactacc cccatggtct aagaccgctt
     1261 caacggatcg gtttggggct tttcttcgga tcaatggcta tggctgtggc tgctttggtc
     1321 gagctcaaac gtcttagaac tgcacacgct catggtccaa cagtcaaaac gcttcctcta
     1381 gggttttatc tactcatccc acaatatctt attgtcggta tcggcgaagc gttaatctac
     1441 acaggacagt tagatttctt cttgagagag tgccctaaag gtatgaaagg gatgagcacg
     1501 ggtctattgt tgagcacatt ggcattaggc tttttcttca gctcggttct cgtgacaatc
     1561 gtcgagaaat tcaccgggaa agctcatcca tggattgccg atgatctcaa caagggccgt
     1621 ctttacaatt tctactggct tgtggccgta cttgttgcct tgaacttcct cattttccta
     1681 gttttctcca agtggtacgt ttacaaggaa aaaagactag ctgaggtggg gattgagttg
     1741 gatgatgagc cgagtattcc aatgggtcat agcggccgcg agaacctgta cttccaatct
     1801 ggttcgatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     1861 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     1921 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     1981 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     2041 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     2101 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     2161 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     2221 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     2281 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     2341 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     2401 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     2461 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     2521 aagtcgtcgc atcatcatca tcatcatcat cattaa
//
