LOCUS       sequence-201367-        4505 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of nucleus-targeting red fluorescent 
            genetically encoded Ca2+ indicator for optical imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4505)
  AUTHORS   Zhao Y, Araki S, Wu J, Teramoto T, Chang YF, Nakano M, Abdelfattah 
            AS, Fujiwara M, Ishihara T, Nagai T, Campbell RE
  TITLE     An Expanded Palette of Genetically Encoded Ca2+ Indicators.
  JOURNAL   Science. 2011 Sep 30;333(6051):1888-91. doi: 10.1126/science.1208592
  PUBMED    21903779
REFERENCE   2  (bases 1 to 4505)
  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"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2011-09-30- 30"; volume: "333"; 
            issue: "6051"; pages: "1888-91"
COMMENT     Sequence Label: CMV-NLS-R-GECO
FEATURES             Location/Qualifiers
     source          1..4505
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(43..62)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        234..613
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        614..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     768..788
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     862..881
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        862..880
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             921..941
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             945..965
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             981..1037
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     1733..1752
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1776..2219
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTFDFPEFLTMMARKMNDTDSEEEIREAFRVFDKDGNGYIGAAEL
                     RHVMTDLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK"
     promoter        complement(2280..2298)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2281..2298)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2318..2335)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2324..2548
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2749..3337)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2829..2848)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3508..4299,4300..4368))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4131..4150
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4369..4473)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
       61 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
      121 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
      181 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
      241 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      301 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      361 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      421 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      481 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      541 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      601 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      661 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      721 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      781 taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac
      841 tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag cttggtaccg
      901 agctcggatc caccatggac ccaaaaaaga agcggaaggt cgatcctaag aagaaacgca
      961 aggtgatggt cgactcatca cgtcgtaagt ggaataaggc aggtcacgca gtcagagcta
     1021 taggtcggct gagctcaccc gtggtttccg agcggatgta ccccgaggac ggcgccctga
     1081 agagcgagat caagaagggg ctgaggctga aggacggcgg ccactacgcc gccgaggtca
     1141 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacatc gtcgacatca
     1201 agttggacat cgtgtcccac aacgaggact acaccatcgt ggaacagtgc gaacgcgccg
     1261 agggccgcca ctccaccggc ggcatggacg agctgtacaa gggaggtaca ggcgggagtc
     1321 tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg
     1381 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     1441 gcccctacga ggcctttcag accgctaagc tgaaggtgac caagggtggc cccctgccct
     1501 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac attaagcacc
     1561 cagccgacat ccccgactac ttcaagctgt ccttccccga gggcttcagg tgggagcgcg
     1621 tgatgaactt cgaggacggc ggcattattc acgttaacca ggactcctcc ctgcaggacg
     1681 gcgtattcat ctacaaggtg aagctgcgcg gcaccaactt cccccccgac ggccccgtaa
     1741 tgcagaagaa gaccatgggc tgggaggcta cgcgtgacca actgactgaa gagcagatcg
     1801 cagaatttaa agaggctttc tccctatttg acaaggacgg ggatgggacg ataacaacca
     1861 aggagctggg gacggtgatg cggtctctgg ggcagaaccc cacagaagca gagctgcagg
     1921 acatgatcaa tgaagtagat gccgacggtg acggcacatt cgacttccct gagttcctga
     1981 cgatgatggc aagaaaaatg aatgacacag acagtgaaga ggaaattaga gaagcgttcc
     2041 gcgtgtttga taaggacggc aatggctaca tcggcgcagc agagcttcgc cacgtgatga
     2101 cagaccttgg agagaagtta acagatgagg aggttgatga aatgatcagg gtagcagaca
     2161 tcgatgggga tggtcaggta aactacgaag agtttgtaca aatgatgaca gcgaagtagg
     2221 aattctgcag atatccatca cactggcggc cgctcgagca tgcatctaga gggccctatt
     2281 ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc
     2341 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2401 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2461 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2521 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     2581 agtccggaac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2641 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2701 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2761 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2821 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2881 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2941 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3001 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3061 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3121 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3181 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3241 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3301 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3361 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3421 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3481 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3541 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3601 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3661 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3721 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3781 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3841 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3901 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3961 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4021 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4081 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4141 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4201 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4261 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4321 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4381 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4441 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     4501 cgtcg
//
