LOCUS       sequence-487454-        2346 bp    DNA     linear   UNA 29-SEP-2025
DEFINITION  mammalian expression of NLS-MARIO, a genetically-encoded fluorescent
            Mg2+ sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2346)
  AUTHORS   Maeshima K, Matsuda T, Shindo Y, Imamura H, Tamura S, Imai R, 
            Kawakami S, Nagashima R, Soga T, Noji H, Oka K, Nagai T
  TITLE     A Transient Rise in Free Mg(2+) Ions Released from ATP-Mg Hydrolysis
            Contributes to Mitotic Chromosome Condensation.
  JOURNAL   Curr Biol. 2018 Feb 5;28(3):444-451.e6. doi: 
            10.1016/j.cub.2017.12.035. Epub 2018 Jan 18.
  PUBMED    29358072
REFERENCE   2  (bases 1 to 2346)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 29, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cub.2017.12.035"; journalName: "Curr Biol"; date: 
            "2018-02-5- 5"; volume: "28"; issue: "3"; pages: "444-451.e6"
COMMENT     Sequence Label: pPB-PGKneo-EF1a-NLS-MARIO-SV40 pA
FEATURES             Location/Qualifiers
     source          1..2346
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             19..39
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(61..63,64..66,67..579)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIE"
     CDS             1348..2082
                     /codon_start=1
                     /product="circularly permuted form of the Venus yellow 
                     fluorescent protein (Nagai et al., 2004)"
                     /label=cp173Venus
                     /note="efficient FRET acceptor for CFP"
                     /translation="MDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRD
                     HMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFTAVVPILVELDGDVNGHKFSVSG
                     EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
                     VYITADKQKNGIKANFKIRHNIE"
     polyA_signal    2219..2340
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2256..2275)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2310..2329
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 gctagcatgg gggggcctcc aaaaaagaag agaaaggtag aagaccccag gatcgatccc
       61 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      121 ggcgacgtaa acggccacag gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      181 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      241 ctcgtgacca ccctgacctg gggcgtgcag tgcttcagcc gctaccccga ccacatgaag
      301 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg taccatcttc
      361 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      421 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      481 aagctggagt acaactacat cagccacaac gtctatatca ccgccgacaa gcagaagaac
      541 ggcatcaagg cccacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      601 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      661 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      721 ctgctggagt tcgtgaccgc cgcccgcatg ctgagcctgg caacccgccc ggaactggaa
      781 gacatcgaag catcggcacg tttctttgaa gacgacgacg gcctgcatga tcactccttc
      841 ttcttctttg aagatgcgga agatcacgcc ggtaactcca ctgtggcatt taccatccgt
      901 gatggtcgtc tgtttactct gcgtgagcgt gaactgcccg cttttcgtct gtatcgtatg
      961 cgtgcccgta gccagtcgat ggtagacggt aacgcctacg agttgctgct ggatctgttc
     1021 gaaaccaaaa tcgaacagtt ggcagatgaa attgaaaata tctatagcga cctggagcag
     1081 ttgagccggg tgattatgga agggcatcag ggcgatgagt acgacgaggc gctctccact
     1141 ctggcggaac tggaagatga cggctgggcc gttgccctgt gtctgatgga tacccagcgc
     1201 gcgctcaact tcctggtgcg taaagcgcgt ttaccgggtg ggcaactgga gcaggcgcgt
     1261 gaaatcctgc gagatatcga atccctgctg ccgcataacg aatccctgtt ccagaaggtg
     1321 aacttcctga tgcaggcggc tgagctcatg gacggcggcg tgcagctcgc cgaccactac
     1381 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1441 taccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1501 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg tggcagcggt
     1561 ggcatggtga gcaagggcga ggagctgttc accgcggtgg tgcccatcct ggtcgagctg
     1621 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1681 tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc
     1741 accctcgtga ccaccctggg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg
     1801 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     1861 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1921 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1981 cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag
     2041 aacggcatca aggccaactt caagatccgc cacaacatcg agtaagaatt gatccaccgg
     2101 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     2161 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     2221 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     2281 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     2341 cccggg
//
