LOCUS       sequence-339177-        7262 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Detect the real-time ER calcium store and local calcium microdomains
            in live cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7262)
  AUTHORS   Luo C, Wang H, Liu Q, He W, Yuan L, Pingyong Xu P
  TITLE     A genetically encoded ratiometric calcium sensor enables 
            quantitative measurement of the local calcium microdomain in the 
            endoplasmic reticulum
  JOURNAL   Biophys Rep (2019) 5, 31–42
REFERENCE   2  (bases 1 to 7262)
  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: "Biophys Rep
            (2019) 5, 31–42"
COMMENT     Sequence Label: GCEPIA1-SNAPER
FEATURES             Location/Qualifiers
     source          1..7262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        206..509
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        510..713
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     664..684
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     710..734
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          862..1240
                     /label=chimeric intron
                     /note="fusion between human cytomegalovirus intron A and 
                     human beta-globin intron 2"
     sig_peptide     join(1322..1366,1449..1460)
                     /codon_start=1
                     /product="modified signal sequence from the mouse 
                     immunoglobulin heavy chain variable region"
                     /label=IgH signal sequence
                     /translation="MGWSCIILFLVATATGAHS"
     CDS             join(1322..1367,1450..1460)
                     /codon_start=1
                     /product="signal sequence from the mouse immunoglobulin 
                     heavy chain variableregion"
                     /label=IgH signal sequence
                     /translation="MGWSCIILFLVATATGAHS"
     CDS             1497..1553
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     1766..1787
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2154..2173)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2289..2729
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKDLGTVLRSLGQNPTEA
                     ELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVWDKDGNGYISAADL
                     RHVMTNLGEKLTDEEVDEMIREADIDGEGQVNYEEFVQMMTAK"
     CDS             2748..3290
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="DKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVEV
                     PAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLKV
                     VKFGEVISYSHLAALAGNPAATAAVKTALSGXPVPILIPCHRVVQGDLDVGGYEGGLAV
                     KEWLLAHEGHRLGKPGLG"
     polyA_signal    3355..3831
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3554..3573)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    3876..3993
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4041..4255
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     4176..4195
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     polyA_signal    4277..4411
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4301..4320)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4355..4374
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4525..4665
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4525..4654
                     /label=AAV2 ITR
     rep_origin      4740..5195
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4827..4846)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5037..5058
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5212..5231)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5331..5353
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5391..5409)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5477..5581
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5582..5650,5651..6442)
                     /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(5800..5819)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6613..7201
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7102..7121
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtctagtt attaatagta atcaattacg gggtcattag
      181 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      241 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      301 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      361 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      421 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      481 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      541 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      601 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      661 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
      721 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
      781 gggaccgatc cagcctccgc ggattcgaat cccggccggg aacggtgcat tggaacgcgg
      841 attccccgtg ccaagagtga cgtaagtacc gcctatagag tctataggcc cacaaaaaat
      901 gctttcttct tttaatatac ttttttgttt atcttatttc taatactttc cctaatctct
      961 ttctttcagg gcaataatga tacaatgtat catgcctctt tgcaccattc taaagaataa
     1021 cagtgataat ttctgggtta aggcaatagc aatatttctg catataaata tttctgcata
     1081 taaattgtaa ctgatgtaag aggtttcata ttgctaatag cagctacaat ccagctacca
     1141 ttctgctttt attttatggt tgggataagg ctggattatt ctgagtccaa gctaggccct
     1201 tttgctaatc atgttcatac ctcttatctt cctcccacag ctcctgggca acgtgctggt
     1261 ctgtgtgctg gcccatcact ttggcaaaga attgggattc gaacatcgat tgaattccac
     1321 catgggatgg agctgtatca tcctcttctt ggtagcaaca gctacaggta aggggttaac
     1381 agtagcaggc ttgaggtctg gacatatata tgggtgacaa tgacatccac tttgcctttc
     1441 tctccacagg cgcgcactcc caggtccaac tgcagggatc catggtcgac tcttcacgtc
     1501 gtaagtggaa taagacaggt cacgcagtca gagctatagg tcggctgagc tcactcgaga
     1561 acgtctatat caaggccgac gagcagaaga acggcatcaa ggcgtacttc aagatccgcc
     1621 acaacatcga gggcggcggc gtgcagctcg cctaccacta ccagcagaac acccccatcg
     1681 gcgacggccc cgtgctgctg cccgacaacc actacctgag cgtgcagtcc atactttcga
     1741 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1801 tcactctcgg catggacgag ctgtacaagg gcggtaccgg agggagcatg gtgagcaagg
     1861 gcgaggagct gttcaccggg gtggtgccca tccaggtcga gctggacggc gacgtaaacg
     1921 gccacaagtt cagcgtgtcc ggcgagggtg agggcgatgc cacctacggc aagctgaccc
     1981 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     2041 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     2101 tcaagtccgc catgcccgaa ggctacatcc aggagcgcac catcttcttc aaggacgacg
     2161 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     2221 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     2281 acacgcgtga ccaactgact gaagagcaga tcgcagaatt taaagaggct ttctccctat
     2341 ttgacaagga cggggatggg acgataacaa ccaaggatct ggggacggtg ctgcggtctc
     2401 tggggcagaa ccccacagaa gcagagctcc aggacatgat caatgaagta gatgccgacg
     2461 gtaatggcac aatcgacttc cctgagttcc tgacaatgat ggcaagaaaa atgaaagaca
     2521 cagacagtga agaagaaatt cgcgaagcgt tccgtgtgtg ggataaggat ggcaatggct
     2581 acatctctgc agcagacctt cgccacgtga tgacaaacct tggagagaag ttaacagatg
     2641 aagaggttga tgaaatgatc agggaagcag atatcgatgg agaaggtcag gtaaactacg
     2701 aagagtttgt acaaatgatg acagcgaagg gaggcggttc agggggcgac aaagactgcg
     2761 aaatgaagcg caccaccctg gatagccctc tgggcaagct ggaactgtct gggtgcgaac
     2821 agggcctgca ccgtatcatc ttcctgggca aaggaacatc tgccgccgac gccgtggaag
     2881 tgcctgcccc agccgccgtg ctgggcggac cagagccact gatgcaggcc accgcctggc
     2941 tcaacgccta ctttcaccag cctgaggcca tcgaggagtt ccctgtgcca gccctgcacc
     3001 acccagtgtt ccagcaggag agctttaccc gccaggtgct gtggaaactg ctgaaagtgg
     3061 tgaagttcgg agaggtcatc agctacagcc acctggccgc cctggccggc aatcccgccg
     3121 ccaccgccgc cgtgaaaacc gccctgagcg gawatcccgt gcccattctg atcccctgcc
     3181 accgggtggt gcagggcgac ctggacgtgg ggggctacga gggcgggctc gccgtgaaag
     3241 agtggctgct ggcccacgag ggccacagac tgggcaagcc tgggctgggt gggggctcaa
     3301 gcgagaagga cgagctgtaa gcttgcctcg agcagcgctg ctcgagagat ctacgggtgg
     3361 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc
     3421 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat
     3481 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt
     3541 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact
     3601 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga
     3661 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc
     3721 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct
     3781 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg
     3841 taggtaaccg gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
     3901 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     3961 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     4021 ccagtttggt taattctagc gaacgctgac gtcatcaacc cgctccaagg aatcgcgggc
     4081 ccagtgtcac taggcgggaa cacccagcgc gcgtgcgccc tggcaggaag atggctgtga
     4141 gggacagggg agtggcgccc tgcaatattt gcatgtcgct atgtgttctg ggaaatcacc
     4201 ataaacgtga aatgtctttg gatttgggaa tcttataagt tctgtatgag accacctcga
     4261 gtctagataa tacgtagatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     4321 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     4381 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     4441 tcaggttcag ggggaggtgt gggaggtttt ttaattcttg cggccgcggc ggtaccggta
     4501 accacgtgcg gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc
     4561 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     4621 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt
     4681 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     4741 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4801 tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4861 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4921 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     4981 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5041 actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata
     5101 agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5161 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     5221 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     5281 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     5341 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     5401 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     5461 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5521 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5581 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5641 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5701 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5761 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     5821 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     5881 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5941 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     6001 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6061 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6121 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     6181 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6241 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6301 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6361 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6421 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     6481 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6541 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6601 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6661 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6721 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6781 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6841 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6901 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6961 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     7021 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7081 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     7141 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     7201 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     7261 gt
//
