LOCUS       sequence-390987-        6094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vivo calcium indicator. Presence of calcium (Ca2+) increases 
            reporter signal intensity. Based on GCaMP7, improved SNR, higher 
            baseline fluorescence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6094)
  AUTHORS   Nava S, Palma W, Wan X, Oh JY, Gharib S, Wang H, Revanna JS, Tan M, 
            Zhang M, Liu J, Chen CH, Lee JS, Perry B, Sternberg PW
  TITLE     A cGAL-UAS bipartite expression toolkit for Caenorhabditis elegans 
            sensory neurons.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Dec 19;120(51):e2221680120. doi: 
            10.1073/pnas.2221680120. Epub 2023 Dec 14.
  PUBMED    38096407
REFERENCE   2  (bases 1 to 6094)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2023-12-19"; volume: "120(51):e2221680120. 
            doi"; pages: " 10.1073/pnas.2221680120. Epub 2023 Dec 14"
COMMENT     Sequence Label: pHW582
FEATURES             Location/Qualifiers
     source          1..6094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    36..130
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     protein_bind    169..263
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     protein_bind    302..396
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     CDS             join(847..855,856..873,874..876,877..909,910..912,913..936,
                     937..963,964..1020,1021..1026,1027..1296,1297..1314,
                     1315..1749,1750..1755,1756..2199)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNPPDQLTEEQIAEFKELFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMYARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      943..952
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1233..1254
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1360..1381)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1621..1640)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     promoter        3839..3943
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3944..4012,4013..4804)
                     /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(4162..4181)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4975..5563
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5464..5483
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5717..5734
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5851..5872
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5887..5892,5893..5910,5911..5917)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5925..5941
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5930..5952
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5949..5965
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5949..5965
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcta gcccatgatt acgccaagct acgggcggag tactgtcctc cgagcggagt
       61 actgtcctcc gagcggagta ctgtcctccg agcggagtac tgtcctccga gcggagtact
      121 gtcctccgag cggagactcc ctagcccatg attacgccaa gctacgggcg gagtactgtc
      181 ctccgagcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag tactgtcctc
      241 cgagcggagt actgtcctcc gagcggagac tccctagccc atgattacgc caagctacgg
      301 gcggagtact gtcctccgag cggagtactg tcctccgagc ggagtactgt cctccgagcg
      361 gagtactgtc ctccgagcgg agtactgtcc tccgagcgga gactccctag gggcggccgc
      421 actgactggg ccggccaaaa atgcataagg ttttgctggc actgttcttt atctttctgg
      481 caccagcatc cgcactggca gtctccgaac cggcctgcag gtcgattttt tgcaaattac
      541 gagcgttgta gggggcggag cgataggtcc tataggtttt ggtatatcat cattcattca
      601 ttcattggta cattcattta cccaccttcc tctttctgag cttctctgga gttctgtgct
      661 tcctttttcc cttatcttta tactgtaatt tttaactttc aggcattgat tggcgcgcct
      721 ctagaggatc cccgggattg gccaaaggac ccaaaggtat gtttcgaatg atactaacat
      781 aacatagaac attttcagga ggacccttgg ctagcgtcga cggtaccggt gcgatcgcta
      841 gcaaaaatgg gttctcatca tcatcatcat catggtatgg ctagcatgac tggtggacag
      901 caaatgggtc gggatctgta cgacgatgac gataaggatc tcgccaccat ggtcgactca
      961 tcacgtcgta agtggaataa gacaggtcac gcagtcagag ctataggtcg gctgagctca
     1021 ctcgagaacg tctatatcaa ggccgacaag cagaagaacg gcatcaaggc gaacttcaag
     1081 atccgccaca acatcgagga cggcggcgtg cagctcgcct accactacca gcagaacacc
     1141 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcgt gcagtccaaa
     1201 ctttcgaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1261 gccgggatca ctctcggcat ggacgagctg tacaagggcg gtaccggagg gagcatggtg
     1321 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1381 gtaaacggcc acaagttcag cgtgtccggc gagggtgagg gcgatgccac ctacggcaag
     1441 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1501 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     1561 gacttcttca agtccgccat gcccgaaggc tacatccagg agcgcaccat cttcttcaag
     1621 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1681 cgcatcgagc ttaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1741 gagtacaacc cacctgacca actgactgaa gagcagatcg cagaatttaa agagcttttc
     1801 tccctatttg acaaggacgg ggatgggaca ataacaacca aggagctggg gacggtgatg
     1861 cggtctctgg ggcagaaccc cacagaagca gagctgcagg acatgatcaa tgaagtagat
     1921 gccgacggtg acggcacaat cgacttccct gagttcctga caatgtacgc aagaaaaatg
     1981 aaatacaggg acacggaaga agaaattaga gaagcgttcg gtgtgtttga taaggatggc
     2041 aatggctaca tcagtgcagc agagcttcgc cacgtgatga caaaccttgg agagaagtta
     2101 acagatgaag aggttgatga aatgatcagg gaagcagaca tcgatgggga tggtcaggta
     2161 aactacgaag agtttgtaca aatgatgaca gcgaagtaac ctgcaggttt tgaattcgct
     2221 gtctcatcct actttcacct agttaactgc ttgtcttaaa atctatgctt ctctttagta
     2281 tctaaaattt tcctagaagc ttacaagtat ataaatggtc tcttctcaat aaaggttgta
     2341 tatttattca tcttattgaa tctgccattt cctcgttttt gcgagtttat ataccttcca
     2401 attttctttc tattgtattt tcaacttcta attttaattc agggaaactg ctgtaccggt
     2461 agaaaaaatg gtctccgagc tcattaaaga aaacatgcat atgaagctct acatggaggg
     2521 aaccgtcaac aaccaccact tcaagtgcac ctccgaggga gagggaaagc catacgaggg
     2581 aacccaaacc atgcgtatca aggccgtcga gggaggacca ctcccattcg ccttcgacat
     2641 cctcgccacc tccttcatgt acggatccaa ggtaagttta aacatatata tactaactaa
     2701 ccctgattat ttaaattttc agaccttcat caaccacacc caaggaatcc cagacttctt
     2761 caagcaatcc ttcccagagg gattcacctg ggagcgtgtc accacctacg aggacggagg
     2821 agtcctcacc gccacccaag acacctccct ccaagacgga tgcctcatct acaacgtcaa
     2881 ggtaagttta aacagttcgg tactaactaa ccatacatat ttaaattttc agatccgtgg
     2941 agtcaacttc ccatccaacg gaccagtcat gcaaaagaag accctcggat gggaggcctc
     3001 caccgagacc ctctacccag ccgacggagg acttgaggga cgtgccgaca tggccctcaa
     3061 gctcgtcgga ggaggacacc tcatctgcaa cctcaaggta agtttaaaca tgattttact
     3121 aactaactaa tctgatttaa attttcagac cacctaccgt tccaagaagc cagccaagaa
     3181 cctcaagatg ccaggagtct actacgtcga ccgtcgtctt gagcgtatca aggaggccga
     3241 caaggagacc tacgtcgagc aacacgaggt cgccgtcgcc cgttactgcg acctcccatc
     3301 caagctcgga caccgttaat tcgctagccg gccatacaag taatccggat gatcgacgcc
     3361 aacgtcgttg aattttcaaa ttttaaatac tgaatatttg ttttttttcc tattatttat
     3421 ttattctctt tgtgtttttt ttcttgcttt ctaaaaaatt aattcaatcc aaatctaaac
     3481 attttttttt ctctttccgt ctcccaattc gtattccgct cctctcatct gaacacaatg
     3541 tgcaagttta tttatcttct cgctttcatt tcattaggac gtggggggaa ttggtggaag
     3601 ggggaaacac acaaaaggat gatggaaatg aaataaggac acacaatatg caacaacatt
     3661 caattcagaa atatggagga aggtttaaaa gaaaacataa aaatatatag aggaggaagg
     3721 aaaactagta aaaaataagc aaagaaatta ggcgaacgat gagaattgtc ctcgcttggg
     3781 cccaaaaggc cttttttttt tggcgccgac gtcaggtggc acttttcggg gaaatgtgcg
     3841 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3901 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3961 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4021 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4081 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4141 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4201 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4261 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4321 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4381 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4441 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4501 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4561 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4621 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4681 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4741 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4801 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4861 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4921 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4981 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5041 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5101 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5161 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5221 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5281 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5341 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5401 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5461 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5521 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5581 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5641 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     5701 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     5761 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     5821 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     5881 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     5941 atttcacaca ggaaacagct atgaccatga ttacgccaag ctgtaagttt aaacatgatc
     6001 ttactaacta actattctca tttaaatttt cagagcttaa aaatggctga aatcactcac
     6061 aacgatggat acgctaacaa cttggaaatg aaat
//
