LOCUS       sequence-353442-        4007 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses the FRET membrane voltage sensor Mermaid in vertebrate 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4007)
  TITLE     CAG-Mermaid in Tol2
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4007)
  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: 
            "Unpublished"
COMMENT     Sequence Label: CAG-Mermaid in Tol2
FEATURES             Location/Qualifiers
     source          1..4007
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    join(71..82,83..245)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     misc_feature    289..360
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     enhancer        367..670
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        672..949
                     /label=chicken beta-actin promoter
     primer_bind     1205..1224
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1280..1289
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2042..2719
                     /codon_start=1
                     /product="monomeric variant of Umikinoko-Green fluorescent 
                     protein"
                     /label=mUkG1
                     /translation="MSVIKEEMKIKLHMEGNVNGHAFVIEGDGKGKPYDGTQTLNLTVK
                     EGAPLPFSYDILTNAFQYGNRAFTKYPADIPDYFKQTFPEGYSWERTMSYEDNAICNVR
                     SEISMEGDCFIYKIRFDGKNFPPNGPVMQKKTLKWEPSTEMMYVRDGFLMGDVNMALLL
                     EGGGHHRCDFKTSYKAKKVVQLPDAHKIDHRIEILSHDRDYSKVKLYENAVARNSLLPS
                     QASK"
     CDS             2729..3385
                     /codon_start=1
                     /product="humanized monomeric Kusabira-Orange fluorescent 
                     protein"
                     /label=hmKO
                     /note="mammalian codon-optimized"
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     polyA_signal    3399..3533
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3436..3455)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3490..3509
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    3541..3596
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     misc_feature    3606..3677
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     protein_bind    complement(3678..3711)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    join(3769..3956,3957..3968)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
ORIGIN
        1 tgaatatctg ttcagacacc catatccact ctgttccaca caggtcagag gtttgtccag
       61 gagttcttga cagaggtgta aaaagtactc aaaaatttta ctcaagtgaa agtacaagta
      121 cttagggaaa attttactca attaaaagta aaagtatctg gctagaatct tacttgagta
      181 aaagtaaaaa agtactccat taaaattgta cttgagtatt aaggaagtaa aagtaaaagc
      241 aagaaagaaa actagagatt cttgtttaag cttgctagca gatatcatca tcttggacca
      301 ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc
      361 cctaggcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      421 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      481 acgtcaatgg gtggactatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      541 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      601 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      661 tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc
      721 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg
      781 gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag
      841 gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc
      901 gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc
      961 gacgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct
     1021 gactgaccgc gttactccca caggtgagcg ggcgggacgg ccttctcctc cgggctgtaa
     1081 ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg
     1141 gctccgggag ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     1201 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt ttggaactct
     1261 aattctccaa attcaagcag ccaccatgga gggattcgac ggttcagatt ttagtcctcc
     1321 agctgattta gttggcgttg acggtgcagt catgcggaac gtcgttgacg tcacgataaa
     1381 tggtgacgtc actgctccgc cgaaagctgc gccaagaaaa agtgaatcgg taaagaaagt
     1441 tcattggaat gatgtagacc aaggaccgag tgaaaaacca gagacaagac aggaggaacg
     1501 aatagatata cccgagattt caggtctatg gtggggcgag aatgaacatg gagtggacga
     1561 tgggagaatg gagataccta ctactggtgt aggtcgcgtc cagtttcgtg tccgagcagt
     1621 gattgatcat ctagggatgc gagtctttgg agtcttccta attttcttgg acatcatcct
     1681 catgatcatt gatctcagtc ttccaggaaa aagtgaatct tcccaatcct tttatgacgg
     1741 gatggctttg gctctttctt gttatttcat gctggattta ggattaagga tatttgcata
     1801 cgggcccaag aatttcttca ccaacccctg ggaggttgct gatggtttga ttatcgttgt
     1861 cacattcgtc gtcacgatat tttacactgt gttagatgaa tacgtgcaag aaacaggagc
     1921 cgatggtttg gggcagttgg ttgtgttggc ccgtttgctg cgtgtggttc gattagcaag
     1981 aatattttat tcccatcaac aaatgaaggc ttcaagcaga agaacaatat caggtgatcc
     2041 tatgagtgtg attaaagaag aaatgaagat caagctgcac atggaaggaa atgtaaacgg
     2101 tcatgcattt gtgattgaag gagatggaaa aggaaagcct tacgatggga cacagacttt
     2161 aaacctgaca gtgaaagaag gcgcacctct ccctttttct tacgacatct tgacaaatgc
     2221 gttccagtac ggaaatagag cattcactaa atatccagcc gatataccag actatttcaa
     2281 gcagacgttt cccgaggggt attcatggga aagaaccatg agttatgaag acaacgccat
     2341 ttgcaacgtg agaagcgaga tcagcatgga aggcgactgc tttatctata aaattcggtt
     2401 tgatggcaag aactttcccc ccaatggtcc agttatgcag aagaaaactt tgaagtggga
     2461 accatccact gagatgatgt acgtgcgtga tgggtttctg atgggtgatg tgaacatggc
     2521 tctgttgctt gaaggaggtg gccatcaccg atgtgacttc aaaacttcct acaaagcgaa
     2581 aaaggttgtg cagttgccag atgctcacaa gattgaccat cgtattgaaa tcttgagcca
     2641 tgacagggat tacagcaaag tcaagctgta tgagaatgcg gttgctcgca attctttgct
     2701 gccaagtcag gcctctaagc ttgagactat ggtgagcgtg atcaagcccg agatgaagat
     2761 gaggtactac atggacggct ccgtcaatgg gcatgagttc acaatcgagg gtgagggcac
     2821 aggcagacct tacgagggac atcaggagat gacactgcgc gtcacaatgg ccgagggcgg
     2881 gccaatgcct ttcgccttcg acctggtgtc ccacgtgttc tgttacggcc acagagtttt
     2941 tactaaatat ccagaagaga tcccagacta tttcaagcag gcctttcctg agggcctgtc
     3001 ctgggagagg tccctggagt tcgaggacgg cggctccgcc tccgtgagcg cccacatcag
     3061 cctgaggggc aacaccttct accacaagtc caagttcacc ggcgtgaact tccccgccga
     3121 cggccccatc atgcagaacc agagcgtgga ctgggagccc tccaccgaga agatcaccgc
     3181 cagcgacggc gtgctgaagg gcgacgtgac catgtacctg aagctggagg gcggcggcaa
     3241 ccacaagtgc cagttcaaga ccacctacaa ggccgccaag gagatcctgg agatgcccgg
     3301 cgatcactac atcggccaca ggctggtgag gaagaccgag ggcaacatca ccgagcaggt
     3361 ggaggacgcc gtggcccact cctaagaaca actagtaaaa cttgtttatt gcagcttata
     3421 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3481 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaggtacc
     3541 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactgc
     3601 agtaccattc ctttaatggt cttttctgga caccactagg ggtcagaagt agttcatcaa
     3661 actttcttcc ctccctaata acttcgtata gcatacatta tacgaagtta tgattccttg
     3721 agcaagcgca attcaattgg tttggtaata gcaagggaaa atagaatgaa gtgatctcca
     3781 aaaaataagt actttttgac tgtaaataaa attgtaagga gtaaaaagta cttttttttc
     3841 taaaaaaatg taattaagta aaagtaaaag tattgatttt taattgtact caagtaaagt
     3901 aaaaatcccc aaaaataata cttaagtaca gtaatcaagt aaaattactc aagtacttta
     3961 cacctctggt tcttgacccc ctaccttcag caagcccagc atctaga
//
