LOCUS       sequence-394850-        9277 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nuclear Localized Fluorescent Calcium Indicator for pan-neuronal 
            expression in C. Elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9277)
  AUTHORS   Nguyen JP, Shipley FB, Linder AN, Plummer GS, Liu M, Setru SU, 
            Shaevitz JW, Leifer AM
  TITLE     Whole-brain calcium imaging with cellular resolution in freely 
            behaving Caenorhabditis elegans.
  JOURNAL   Proc Natl Acad Sci U S A. 2015 Dec 28. pii: 201507110.
  PUBMED    26712014
REFERENCE   2  (bases 1 to 9277)
  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. 2015 Dec 28. pii: 201507110."
COMMENT     Sequence Label: Prab-3::NLS::GCaMP6s
FEATURES             Location/Qualifiers
     source          1..9277
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4415..4435
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4436..4444,4445..4462,4463..4465,4466..4498,4499..4501,
                     4502..4525,4526..4552,4553..4609,4610..4615,4616..4885,
                     4886..4903,4904..5338,5339..5344,5345..5788)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      4532..4541
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     4822..4843
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4949..4970)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5210..5229)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(6713..6735)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6835..6854
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6936..6954)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7022..7126
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7127..7195,7196..7987)
                     /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(7345..7364)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8158..8746
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8647..8666
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8900..8917
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9034..9055
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9070..9075,9076..9093,9094..9100)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9108..9124
                     /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     9113..9135
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9132..9148
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9132..9148
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggc gagttttgac tggctttcaa aaactgagtg attactaatt
       61 atttttgaaa aaattaacag ttgtgatatg atatttagac atttttgcaa tatataattt
      121 gtttgaaaaa aaaacatttt ttaatggtct tcgaagctat aatagttttt cgtaaaattt
      181 taaataaaac agctgacaga tatccacatt caataattga aagtatgcag ttctcttttt
      241 ttttgtcaaa tagaaaaaaa gtaacagatg gtcagatata atctaggtta atctgactct
      301 ggtcatcgat gaactcattt tcatcataat cattcttctt tcaaatggtc tgttcggaat
      361 tcgaatattc aataaatgga tatcttatta gatgctccgg atttattcca ccaaacttcc
      421 aagacgtaag ttcaatttaa acaaaattac tatagcttga aaaaatgaga aaaaacgagc
      481 agataaaaaa ataacagaaa aaatccataa aaatgacgag ggttccgtgc gaaaatcgtc
      541 attgtttttg gcttgcatca cacttttccg cgagtcagcc acactgtttt atcgattatt
      601 atttttctga tttcagaaac cggaaactta caactgagat gctttcattt tgattttttt
      661 aaatttagtt ttgatttcag cgatttgttt gaatataatt atgcatgagt tggcttttgc
      721 atttttagat aaataagttc acatgctcgg acaaattgtg tggtttaatc tgtgaaagtt
      781 ttgaatcttc tgtttgaaaa aaaaaacatt tttaatttgt atatacgagt gaattggtta
      841 ggccctggtt acgttttcca gcgaattcat tcaaatttct tattttttct attttttcaa
      901 ttaaataatt tagttctcta tcacatgtgt atgatcaaaa tttcattcct aatcgtaagc
      961 aaatttaggc tcatccgtag tataattttg ttatgaaggg cttcacagta aaagctcaaa
     1021 ctccaacatt tagcaactag caaccaacaa aaaaccaaag aaatgtcagt tttgaaaagt
     1081 cgttatttct ttgtcttctc acaattcaca catcatcttc tcgttttcct ccgttttttt
     1141 gtcgtgcttc cttcttttct ttacacattt ttctaacctt tttttaagtc tatcctctcc
     1201 taatcctttg tactctctaa tcagttttgc tggttgtgct gcacaatttt tatcagattt
     1261 atgcgcgaaa caaaagcctc tcgcggagag ctccgccaaa tccggtcttc ttttttcctt
     1321 gcctatcgga tttcacaaac attcagtttg tcacttgacg atgtgttgga ggcattacta
     1381 ctaggaagtg ttattcatat tttttattcg tttctttttg gtgactaaac gaaatattcc
     1441 gaacgttttc ctgatttcag gcggcggaaa cgaagaagac cacgaataat caacaggctg
     1501 ccattgcatc cgccagaagt cgagtgagtt ctttgaaaat ctttattaga gttatttgtg
     1561 acaatttgcc actttttttt taatgtttaa tgaatctgta acgaaattaa taccaagggt
     1621 gtccaacaga actatctgag cggttattta atttttagat agtatatttc ttccgatcgt
     1681 aacatacata ttttcaagca tagtactgtg atttaagtta cagttttcca atcagtcttg
     1741 attttaataa aaacttttca actgcaaaaa tgcctctgaa gtatttttct atatttttat
     1801 agttgaataa ttttttgatt ggcacaaaga ccgccgagat atgagccgtt aaagtaaagc
     1861 tgcatggatt caatatggtt tgtatttgat tggagtcact tacatggaaa tatttgtatt
     1921 taaatctcat attttaaaac aagacaaaca ccaaattaaa tttttattaa tctgaacatt
     1981 tttcaatcga tttggtaaac aaattattag atgtttccct caaaaattgt aacaattcct
     2041 aaattgtggc acttataata acatcgtcct gaattctcaa aaaaaaccga atgctaagag
     2101 agatttcgtt taactgggac ctgtaactga attttcaaat attggcagtt tttgataaac
     2161 tgggagggaa atcaaaaacc aaaactacag tacccctcgt gttataataa attgtcattc
     2221 actccttttt tgttcccaac atttgctccc atctcctgac gagaaagaac aattttatga
     2281 gcacgggaac ccgtgggcgt ccatccgttc ctcctgtgca atttcagccc ccttcctcta
     2341 cattctctgt gttcccgttt ttatcgtttt atgggtatcg ggaacggaca gatcgcttcc
     2401 atttgcttct attccgtcct cacaagacgt cccactaact gattttcttc tgatttcttt
     2461 tcactgaaaa tgggaaagaa agaaaaagac gatgaatcga cgacttcttc tcaggagaag
     2521 gtgaagaaga aggataagga gaaagttcat ctaaggctga aagataaagt aaaaaaactt
     2581 agaaaatcta agtcgattgc agtatcttct ccagctgatt atatgacttg tcagtgtgaa
     2641 ccatgcaaaa aagtagctct cgaggtaata atttttaaac tcaatacgta gttaagtttg
     2701 aactcctttt aatttactta tcatctaatt taccgtcaca atagtacttc cattagcaat
     2761 tattttaaaa ttacatgcat agcttcaatg ttcgtttttt tgcgttctca tatcgtatgt
     2821 tgtctgcctc tcgaatactg actgtccgac tagaatgtcc cggtaggatt actgtacggt
     2881 cactgggaat aaactgtttc tgaaatcaat ctgaaattcc gtagatcttc ttggttcaat
     2941 ttcttattcc tcaatctttg gacaacgtca ccttcagagt cgtcgttgag tatcccatca
     3001 ttaactagtc aatctttctc tctttttcat ttcccccaac ttttctgcat attttttgac
     3061 gacgacgacc tcgacggcaa cgagaataca tttatctatt ttttagtata gcatatatac
     3121 attttctact tttctctaac ccatttcaga cgtatagatt ttggttttcg aatgcaactt
     3181 tgagattttt caaagatctt cagatgggag cagtggactg tctatttgat attcttaaaa
     3241 tgtactctat aaaagatgaa actgtaatga ccaattccct acataaatat ttttaactaa
     3301 ttggtcggat atttggggat cagagaagta cttgtttttg cgatattaat aatgatatat
     3361 ttttctagta aaaaaatgtt aagcgatttg tgaataatta agttggcttg aaaaagattt
     3421 tttgatagct atgtattttg ttaatttaaa tcagatttta aaactttatt attgactatt
     3481 tatatatatt atatattata tattttcatg gtttaatcaa aatctattaa attatttcta
     3541 agatctctaa atatcgaatt ctctaaaaag cttcatcaac aaaagcattt caattatcta
     3601 acacaatgag aataattcaa gaactgaaat agaaagaaat ctagaaaaaa aagagagtga
     3661 ggtgccctat taaatcggtg ctctcctctc taacagaagc gcctacacgc acccaagaga
     3721 gctctttttc tttctttttt cggagcacac atacacgcat acactcctca gaagtgtata
     3781 gagaaaatgg gagaagaagc gaaagaagac gaagaagcgg ttgtgatgta tgcatcgggc
     3841 gtttctatcc ctctccgtga gcaacgagct agtcaaccca tcgatccatt tttgtgagca
     3901 cacacagaga gagactcaaa cacactcaca gcttcacact gagctccaag agctctgtag
     3961 agagaagcca gagtatgctt tcctcgattt tcctaacgtc ttcttcttct gcttttttct
     4021 ttttcccaac gagttgtagt ccactaactt caatttttcc tccagagaaa gagttttaga
     4081 gagacagctg ataaatacac tagagaaata aaaagaatga taaaaatttg ctattttaat
     4141 cagttccctc tcgtttctct aagttttgtt gttgggttgg gaaattaaaa ttggaaaatg
     4201 gaaataacgt ttttttctgt ctaatcttga attctccaga catttcttat ttcctttttt
     4261 gttgcaagtt tttcttcctt ctctaatttt taatcaaaga tatctcgcta tgaactttga
     4321 atttttgata ttttgattct ataaatacat aaagtaatct aaatgctctt ttaaaataaa
     4381 tctacagtag ccctattttc agggatccat ggctccaaag aagaagcgta aggtaatggg
     4441 ttctcatcat catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg
     4501 ggatctgtac gacgatgacg ataaggatct cgccaccatg gtcgactcat cacgtcgtaa
     4561 gtggaataag acaggtcacg cagtcagagc tataggtcgg ctgagctcac tcgagaacgt
     4621 ctatatcaag gccgacaagc agaagaacgg catcaaggcg aacttccaca tccgccacaa
     4681 catcgaggac ggcggcgtgc agctcgccta ccactaccag cagaacaccc ccatcggcga
     4741 cggccccgtg ctgctgcccg acaaccacta cctgagcgtg cagtccaaac tttcgaaaga
     4801 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     4861 tctcggcatg gacgagctgt acaagggcgg taccggaggg agcatggtga gcaagggcga
     4921 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     4981 caagttcagc gtgtccggcg agggtgaggg cgatgccacc tacggcaagc tgaccctgaa
     5041 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     5101 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     5161 gtccgccatg cccgaaggct acatccagga gcgcaccatc ttcttcaagg acgacggcaa
     5221 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     5281 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacct
     5341 gccggaccaa ctgactgaag agcagatcgc agaatttaaa gaggctttct ccctatttga
     5401 caaggacggg gatgggacaa taacaaccaa ggagctgggg acggtgatgc ggtctctggg
     5461 gcagaacccc acagaagcag agctgcagga catgatcaat gaagtagatg ccgacggtga
     5521 cggcacaatc gacttccctg agttcctgac aatgatggca agaaaaatga aatacaggga
     5581 cacggaagaa gaaattagag aagcgttcgg tgtgtttgat aaggatggca atggctacat
     5641 cagtgcagca gagcttcgcc acgtgatgac aaaccttgga gagaagttaa cagatgaaga
     5701 ggttgatgaa atgatcaggg aagcagacat cgatggggat ggtcaggtaa actacgaaga
     5761 gtttgtacaa atgatgacag cgaagtgagg gcgccatgag ccgtagacga aaagcgaatc
     5821 cgacaaaact gagtgaaaac gcgaagaagc ttgccaagga agttgaaaat taaggtacca
     5881 tggtattgat atctgagctc cgcatcggcc gctgtcatca gatcgccatc tcgcgcccgt
     5941 gcctctgact tctaagtcca attactcttc aacatcccta catgctcttt ctccctgtgc
     6001 tcccaccccc tatttttgtt attatcaaaa aacttctctt aatttctttg ttttttagct
     6061 tcttttaagt cacctctaac aatgaaattg tgtagattca aaaatagaat taattcgtaa
     6121 taaaaagtcg aaaaaaattg tgctccctcc ccccattaat aataattcta tcccaaaatc
     6181 tacacaatgt tctgtgtaca cttcttatgt tttttacttc tgataaattt ttttgaaaca
     6241 tcatagaaaa aaccgcacac aaaatacctt atcatatgtt acgtttcagt ttatgaccgc
     6301 aatttttatt tcttcgcacg tctgggcctc tcatgacgtc aaatcatgct catcgtgaaa
     6361 aagttttgga gtatttttgg aatttttcaa tcaagtgaaa gtttatgaaa ttaattttcc
     6421 tgcttttgct ttttggggtt tcccctattg tttgtcaaga tttcgaggac ggcgtttttc
     6481 ttgctaaaat cacaagtatt gatgagcacg atgcaagaaa gatcggaaga aggtttgggt
     6541 ttgaggctca gtggaaggtg agtagaagtt gataatttga aagtggagta gtgtctatgg
     6601 ggtttttgcc ttaaatgaca gaatacattc ccaatatacc aaacataact gtttctacta
     6661 gtcggccgta cgggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     6721 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     6781 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     6841 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     6901 ggagaaaata ccgcatcagg cggccttaat taagggcctc gtgatacgcc tatttttata
     6961 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     7021 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     7081 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     7141 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     7201 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     7261 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     7321 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     7381 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     7441 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     7501 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     7561 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     7621 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     7681 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     7741 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     7801 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     7861 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     7921 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     7981 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     8041 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     8101 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     8161 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     8221 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     8281 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     8341 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     8401 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     8461 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     8521 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     8581 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     8641 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     8701 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     8761 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     8821 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     8881 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     8941 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     9001 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     9061 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     9121 acaatttcac acaggaaaca gctatgacca tgattacgcc aagctgtaag tttaaacatg
     9181 atcttactaa ctaactattc tcatttaaat tttcagagct taaaaatggc tgaaatcact
     9241 cacaacgatg gatacgctaa caacttggaa atgaaat
//
