LOCUS       sequence-395094-        6234 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Plasmid for ectopic integration and expression of Khd4-Ada-Gfp. The 
            4282 bp khd4-ORF is fused C-terminally to the 1200 bp adar catalytic
            domain (Ada) with three repeats of GGGGS linker in between, fol.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6234)
  AUTHORS   Sankaranarayanan S, Haag C, Petzsch P, Kohrer K, Matuszynska A, 
            Zarnack K, Feldbrugge M
  TITLE     The mRNA stability factor Khd4 defines a specific mRNA regulon for 
            membrane trafficking in the pathogen Ustilago maydis.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Aug 22;120(34):e2301731120. doi: 
            10.1073/pnas.2301731120. Epub 2023 Aug 17.
  PUBMED    37590419
REFERENCE   2  (bases 1 to 6234)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2023-08-22"; volume: "120(34):e2301731120. 
            doi"; pages: " 10.1073/pnas.2301731120. Epub 2023 Aug 17"
COMMENT     Sequence Label: p Pcrg1: Khd4-Ada-Gfp (CbxR)
FEATURES             Location/Qualifiers
     source          1..6234
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(5515..5517,5518..5520,5521..6234)
                     /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(5515..5517,5518..5520,5521..6231)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5560..5581)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5821..5840)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6168..6189
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atggatttct actcgacgtc cttcactatt ccgcgctcca tcggtgccag cagttttgtc
       61 acggagacta ccaatcccaa cggcagcatt acagcagcat cgtcgtcgag cttcagcctc
      121 gaggctcctc gtcctggctc tcatgtaggt gcgcgaactg cgcccacagc ttcggctgct
      181 cccgagccac ctgcctcttc gttgccaggc cgcttcacct cgatgcgctc ccgttttcct
      241 cctagcgtca aggctttcat gcccagtgcg tttggcggca agcgcaccag cgacaggatc
      301 gtcgaggctg tcggcgcaga aaaagcagag gccgacgctg ccaccggctc ctcttctcaa
      361 cctgttaaag ccgaacccga gcagtcggcg ccgctggccc ctatcggcga gaggggaaat
      421 gctcgtcctt ccaagctcgt caatcccagt ctggccaaca tggatgacaa cggtagcagc
      481 cagcatcgtg acagcagatc caccgttggc actcggtcca gctttgacat ttcatcggca
      541 gcctccttca ccgcagacag caccgctgct agtagcgcca tcggcgtcaa ctcgtcgagc
      601 accacccatt cggcttccaa ttcgatctcg gccaaccatg tgcagcagga tcgaggcatc
      661 tcgctttcac agagcagcaa agatcttcaa gaagctatcc agaaactcag ctccgatatc
      721 atggccaatc accagtgcct agtctcgttt gcttcagtgc cgcaaaacga cgagtcgcgc
      781 atgcccccaa ccccgctctc gcatcccttt gctcgccaga gctccttctc ttcgttgcag
      841 agcctcgatc aattgccttt tcccaatcca aatggatctt ccggctatcc ctatcagcag
      901 taccatccac agcccatcgg cacgcctgac tcgcactcgt ccgccttctt ccctggacgc
      961 gccgctggag aagcgctcgg ttctggtttc cccggcctca actcaagttc gctctcgctc
     1021 tcgtcgctca atccgcccgc gtctggcagc cacgtcgccc aagaggtcgg tctctcgtcc
     1081 gctcagccgc tccggtcgcc gcagccaaat gccattggtg cttcgcgagc cagcctcagt
     1141 agcatgccca aaccaccgca gtacaacttt catctcagcg gtggctacca acaggtcatg
     1201 aatgctcgag gcaccatctt acgcgaaaat ccgttcaaat cgacctcctc catcaaagtg
     1261 ccgcgcgccg atgtgttcga cgtcgcaccc gcctcgtcgc ctcccgcaaa agagagcatc
     1321 aaggaagatg tgcgtcgcaa actcgatgaa gtacagaccg ccacgggcgc aaccatcacg
     1381 ctcaaaaaca gtgaagtgcg cggcgctgat ctcggctatg gcctcgagac cgagcgcatc
     1441 gtcgaggtgc tcgtctcggg aacctttgaa agtgtcgagt tggcccgggt caaggtgctc
     1501 gtcatcttgg acgagattgc tgggcttcga tccgagacat gcgagatcga ctacaagttg
     1561 cacaacatca tcggcggccg caagcgctgc gttgtgcaga agatcgagga agaaacgggc
     1621 accagcatct acttgccaag ttcttttctc ggcaccttcg gctcgagtct tgccagccgc
     1681 aacgacggca aggcagtcgc ggctcatcaa aatcaaatcc acatcacggg cgaattcttt
     1741 ggcgttcagc gtgcacgcga catgcttttc caggtgtcga tgcacaagag caagggcatc
     1801 atttcacgag atgcagctat cctacctcgc aagctcgatt ggatgctgac cgaccgtctc
     1861 gaggagctcc gatccatcat gatcgacaac ggcacctttg tcgccttccc attgctcggc
     1921 agtcagacca gcgtcatctc ggtctttggc gatcatcgcg ttaacatcga acgcacaatc
     1981 cgatccatca tgcagcttgc gtgccagttc tacgtcgcct cgctctggct cttacccgtc
     2041 ggtttcgacg tctacatgcc atctccacag gctaatctca atccaacgca ggtcgcgccc
     2101 atgctcaagc acgtcgccaa cgcttcgggc gccgaggtgg tgttcaagag caactgcttc
     2161 gagattcacg gtctcgagtc cgaggtgcga accgccgttt cggcgttgct cgaactcgag
     2221 ttgatcaaaa gcttcaactt cgaagtgcgc ttccagattg agctggccaa cgagcaccgt
     2281 gagtttatct cgggcaagaa gaacggcaaa gtcaacaagg tcatgaagca gtgtggcgtg
     2341 cgcatcaagt ttgaaacatt caatgactac aacttcctaa tcgacgtctc gggaaacgat
     2401 cgcaacggag ttttacaagg cctcggtctt ttacaagagg agttgccagc cgagatgtcg
     2461 ttccacgtcc ccgaggcgta ccacaaacgc atcattggcg tcggtggcaa gaacatccaa
     2521 cgcatcatga aaaaatttgg cgtctatgtc aagtttagca atgccgagga gtttgctgcg
     2581 ctcggaggtt acctcgataa cgaagataac gtcatcgcac gaacaccggc caaaaatgct
     2641 gccaaccttg aaaacctcaa gctgtcggtg atggagctgg tcaatcccaa ggacaaggac
     2701 tacatcagtg agacggtcac catctcgcgt cgctaccatc gaactttgct tggcgagaag
     2761 gccatcttca tccatgacat tgagagcaag acaagttcca gcgttcgctt ccctgcgcga
     2821 gagtcagcct cggatctggt gaccatcttc ggacccgagt cgcagattca cattgctgcg
     2881 cagatgcttc tcgaccacgt tccctttgaa gccgagttcc gcgcccccaa ctcgaccgag
     2941 ctcggcgatg ctattcacag tgccgagttt gctgcactcg ctgagcagct caagcgcgac
     3001 ctcagtatct ctgtctcgcc tgtcattgag cagcgacgac ccggtggcga ggccgtcttc
     3061 aagttgcggc tcaacaggag taacaccgac ttcctgccca cggcaaagga tgcaatcgag
     3121 gatttcctca tcaatcgcaa cgccaacatc tacgctgcac cctctcgaac tcgctccgat
     3181 tcgtttgcat cggccttccc gcactttgcc aacaagttga tctcgacggc tggcgcggcc
     3241 gaatcaaatg aatcgttcaa cactgcggcc gcagcggcag ccgccgcggc cgaacaggcg
     3301 cgtgtgaatg agcgcaggct gcgtgctgct gcctcgacac ccgatatcaa agcgctcttt
     3361 gatgcaccgt cccagcacct tcatggcggc ccaggatttg cgtcgagcaa tggcggaagc
     3421 gcagctacag ctgctaactc gcctttgatc acgtctccgc tctacacttc gccgtacggt
     3481 aacggtcgtg ggtttggcag tgatgtgtgg ggcgctccta cacgtgcctt cacagccagt
     3541 aacgctagca acgccacgcc aagcttgatg gggttgcctc cgccttccgt cggaacgcct
     3601 ggcgcaggcc tcgttagtgc agctggacct ggacccatcg gggctggagt cagtgtgcca
     3661 acgagtggcg ggatccagtt tccctcgcag cctagcctgc accagcaaag cggacatcgg
     3721 ctgtcggacg agatgcacat gaaccgaggc ttggagggca tgtcgatgga agaccgcgtc
     3781 aaggcgcttc gaaagcctcg ctcgtttgcg catcgggcac agagtttgga cattggcgcc
     3841 atggcagcgc agcaggcctc gcagcatgca agtggaagca tgagtgttgg cccgtcgacg
     3901 ccttacggtc ttggtcctat cggcacgggt gcccctggca tgtctggcca cttccctggc
     3961 atgacaagct cgcaaggcac ctttggcggt ggtgcacctc accacctcta ccagcctcca
     4021 acaccgcagc agcaggcaca gcagcaactc caataccaac agcagcagca gcaacagcag
     4081 caacagcagc aacagccggg ttacggcatg ccgcatcaac ctcagcactt tcatgcgtct
     4141 tccgctagca tctcgcgcct tccgcccggt cgcaacacac agaaccctga ttcgaccacc
     4201 atggacgagg tgagccgggt gcttgctcaa ctcgcttttg accgggcggc caacgcggcc
     4261 caaggaggcg gtggttccgg tggcggagga tcgggaggtg gcggttcgtc catggtggtt
     4321 cctcaaaaga atgtgccact gcctatcgat gataagtcgt cttctatgga actgcctcag
     4381 atccacgcag ataccattgg ccgactcgtc ctcgaaaagt tcatggaagt gattaaggga
     4441 caggaagcgt attcccgacg caaggtgctg gctggtatcg tcatgaccga gaacatgaat
     4501 ttttgtgagg ctaaggtgat ctctgtctcg accggcacca aatgtgtctc cggagagcac
     4561 atgtcggtca atggagcagt gttgaatgat tcccatgcag aaattgtttc ccgtcgttgt
     4621 ttgctgaaat atctctacgc tcaactggac ctgcaatgca atcaggcaac agcttaccaa
     4681 tcgatcttcg ttcgtaacac agatggacag tatccataca agctcaagag cggtgtgcat
     4741 ttccatctgt acatcaacac agcgccgtgc ggcgacgccc gaattttttc gccccacgaa
     4801 aatgataccg gcgttgataa gcatccaaat cgtaaggcgc gaggccaatt gcgaacaaag
     4861 atcgaatcgg gacaaggcac aatcccggtg aagagcagcg atggcattca gacctgggac
     4921 ggagttctcc aaggacagcg tctgcttacc atgtcgtgct ctgataaaat tgctcgctgg
     4981 aatattgtcg gcatccaggg atcgctgctg tcgtccatca tcgagccggt gtacctccat
     5041 tctatcgttc tgggatcgtt gctgcaccct gagcacatgt accgtgccgt gtgtggccgt
     5101 atcgagaagt cgatccaggg actccctcca ccttaccacc tgaataaacc acgacttgct
     5161 ctcgtcacct cggccgaacc gcgcaatcag gcgaaagcgc ctaactttgg cattaactgg
     5221 accattggag acaccgaact tgaagtggtg aactctctga ctggtcgcac cattggcggc
     5281 caggtttcgc gcatcacgaa gcaagccttt tttgtcaaat atggcttcct gatggcgaac
     5341 ctgcccggta tcctcgtccg taaggttacg acagactatg gccagacgaa agcgaacgtg
     5401 aaagattatc aaattgccaa gctggaactg ttttcggcct ttaaacgcga ggatttggga
     5461 tcgtggctca aaaagccgat cgaacaagat gagttcggac tggcagaatc ccatatggtg
     5521 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     5581 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     5641 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     5701 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     5761 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     5821 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     5881 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     5941 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     6001 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     6061 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     6121 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     6181 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaa
//
