LOCUS       sequence-339086-        7060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TnC-based calcium sensor with green-red FRET pair ratiometric 
            readout.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7060)
  AUTHORS   Zhang D, Redington E, Gong Y
  TITLE     Rational engineering of ratiometric calcium sensors with bright 
            green and red fluorescent proteins.
  JOURNAL   Commun Biol. 2021 Jul 29;4(1):924. doi: 10.1038/s42003-021-02452-z.
  PUBMED    34326458
REFERENCE   2  (bases 1 to 7060)
  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: "Commun 
            Biol."; date: "2021-07-29"; volume: "4(1):924. doi"; pages: " 
            10.1038/s42003-021-02452-z"
COMMENT     Sequence Label: AAV-Camk2-NES-Twitch-NR
FEATURES             Location/Qualifiers
     source          1..7060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        129..1417
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     primer_bind     complement(1640..1658)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1908..1927
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     regulatory      2872..2881
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3167..3755
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3220..3240)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3638..3649)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3787..4263
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3986..4005)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4303..4443
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4303..4432
                     /label=AAV2 ITR
     rep_origin      4518..4973
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4605..4624)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4815..4836
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4990..5009)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5109..5131
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5169..5187)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5255..5359
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5360..5428,5429..6220)
                     /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     complement(5578..5597)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6391..6979
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6880..6899
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(7041..7060,1..110)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag
       61 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgcac
      121 gcgtttaaca ttatggcctt aggtcacttc atctccatgg ggttcttctt ctgattttct
      181 agaaaatgag atgggggtgc agagagcttc ctcagtgacc tgcccagggt cacatcagaa
      241 atgtcagagc tagaacttga actcagatta ctaatcttaa attccatgcc ttgggggcat
      301 gcaagtacga tatacagaag gagtgaactc attagggcag atgaccaatg agtttaggaa
      361 agaagagtcc agggcagggt acatctacac cacccgccca gccctgggtg agtccagcca
      421 cgttcacctc attatagttg cctctctcca gtcctacctt gacgggaagc acaagcagaa
      481 actgggacag gagccccagg agaccaaatc ttcatggtcc ctctgggagg atgggtgggg
      541 agagctgtgg cagaggcctc aggaggggcc ctgctgctca gtggtgacag ataggggtga
      601 gaaagcagac agagtcattc cgtcagcatt ctgggtctgt ttggtacttc ttctcacgct
      661 aaggtggcgg tgtgatatgc acaatggcta aaaagcaggg agagctggaa agaaacaagg
      721 acagagacag aggccaagtc aaccagacca attcccagag gaagcaaaga aaccattaca
      781 gagactacaa gggggaaggg aaggagagat gaattagctt cccctgtaaa ccttagaacc
      841 cagctgttgc cagggcaacg gggcaatacc tgtctcttca gaggagatga agttgccagg
      901 gtaactacat cctgtctttc tcaaggacca tcccagaatg tggcacccac tagccgttac
      961 catagcaact gcctctttgc cccacttaat cccatcccgt ctgttaaaag ggccctatag
     1021 ttggaggtgg gggaggtagg aagagcgatg atcacttgtg gactaagttt gttcgcatcc
     1081 ccttctccaa ccccctcagt acatcaccct gggggaacag ggtccacttg ctcctgggcc
     1141 cacacagtcc tgcagtattg tgtatataag gccagggcaa agaggagcag gttttaaagt
     1201 gaaaggcagg caggtgttgg ggaggcagtt accggggcaa cgggaacagg gcgtttcgga
     1261 ggtggttgcc atggggacct ggatgctgac gaaggctcgc gaggctgtga gcagccacag
     1321 tgccctgctc agaagcccca agctcgtcag tcaagccggt tctccgtttg cactcaggag
     1381 cacgggcagg cgagtggccc ctagttctgg gggcagctct agagcggtac cggatccgcc
     1441 accatgctgc agaacgagct tgctcttaag ttggctggac ttgatattaa caagactgga
     1501 ggaggttctg tgagcaaggg cgaggcagtg atcaaggagt tcatgcggtt caaggtgcac
     1561 atggagggct ccatgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     1621 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttctcc
     1681 tgggacatcc tgtcccctca gttcatgtac ggctccaggg ccttcaccaa gcaccccgcc
     1741 gacatccccg actactataa gcagtccttc cccgagggct tcaagtggga gcgcgtgatg
     1801 aacttcgagg acggcggcgc cgtgaccgtg acccaggaca cctccctgga ggacggcacc
     1861 ctgatctaca aggtgaagct ccgcggcacc aacttccctc ctgacggccc cgtaatgcag
     1921 aagaagacaa tgggctggga agcgtccacc gagcggttgt accccgagga cggcgtgctg
     1981 aagggcgaca ttaagatggc cctgcgcctg aaggacggcg gccgctacct ggcggacttc
     2041 aagaccacct acaaggccaa gaagcccgtg cagatgcccg gcgcctacaa cgtcgaccgc
     2101 aagttggaca tcacctccca caacgaggac tacaccgtgg tggaacagta cgaacgctcc
     2161 gagggccgcc actccgatgc tagcgaagaa gagctatcag agtgctttcg catttttgac
     2221 ttcgatggaa atggtttcat cgacagggag gaatttggag acatcatccg ccttactgga
     2281 gaacagctca cagatgaaga tgtcgatgag atatttggag attcagacac agacaaaaat
     2341 ggaaggattg actttgatga gttcctgaag atggtggaga atgtccagcc tctggccaac
     2401 cgctaccgga gcactgcgcg gaccacctac acctttgcca agccaatggc ggctaactat
     2461 ctgaagaacc agccgatgta cgtgttccgt aagacggagc tcaagcactc catgaccgag
     2521 ctcaacttca aggagtggca aaaggccttt accgatatga tgggcatgga cgagctgtac
     2581 aaggggggca ctggagggag tggaggcggt ggtagtatgg tgagcaaggg cgaggaggat
     2641 aacatggcct ctctcccagc gacacatgag ttacacatct ttggctccat caacggtgtg
     2701 gactttgaca tggtgggtca gggcaccggc aatccaaatg atggttatga ggagttaaac
     2761 ctgaagtcca ccaagggtga cctccagttc tccccctgga ttctggtccc tcatatcggg
     2821 tatggcttcc atcagtacct gccctaccct gacgggatgt cgcctttcca ggccgccatg
     2881 gtagatggct ccggatacca agtccatcgc acaatgcagt ttgaagatgg tgcctccctt
     2941 actgttaact accgctacac ctacgaggga agccacatca aaggagaggc ccaggtgatg
     3001 gggactggtt tccctgctga cggtcctgtg atgaccaaca cgctgaccgc tgcggactgg
     3061 tgcatgtcga agaagactta ccccaacgac aaaaccatca tcagtacctt taagtggagt
     3121 tacaccactg taaatggcta atgaattcga tatcaagctt atcgataatc aacctctgga
     3181 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3241 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3301 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3361 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3421 caccacctgt cagctccttt ccggaacttt cgctttcccc ctccctattg ccacggcgga
     3481 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3541 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct atgttgccac
     3601 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3661 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3721 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag
     3781 atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc
     3841 actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag
     3901 gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg
     3961 aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa
     4021 tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc
     4081 gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag
     4141 agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac
     4201 ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc
     4261 cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     4321 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     4381 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     4441 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     4501 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4561 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     4621 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4681 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     4741 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4801 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     4861 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     4921 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     4981 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     5041 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     5101 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     5161 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     5221 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5281 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5341 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5401 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5461 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5521 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5581 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5641 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     5701 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5761 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5821 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5881 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5941 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     6001 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6061 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6121 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6181 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6241 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6301 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6361 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6421 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6481 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6541 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6601 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6661 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6721 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6781 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6841 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6901 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6961 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     7021 ctggcctttt gctcacatgt cctgcaggca gctgcgcgct
//
