LOCUS       sequence-229565-        8917 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For Calcium imaging, expression of G-CaMP6 under the expression of a
            npr-4 promoter (AVA and RIV interneurons).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8917)
  AUTHORS   Hori S, Oda S, Suehiro Y, Iino Y, Mitani S
  TITLE     OFF-responses of interneurons optimize avoidance behaviors depending
            on stimulus strength via electrical synapses.
  JOURNAL   PLoS Genet. 2018 Jun 25;14(6):e1007477. doi: 
            10.1371/journal.pgen.1007477. eCollection 2018 Jun.
  PUBMED    29939997
REFERENCE   2  (bases 1 to 8917)
  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: "PLoS 
            Genet."; date: "2018-06-25"; volume: "14(6):e1007477. doi"; pages: "
            10.1371/journal.pgen.1007477. eCollection 2018 Jun"
COMMENT     Sequence Label: pPD_Pnpr-4::G-CaMP6s
FEATURES             Location/Qualifiers
     source          1..8917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     145..162
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    279..300
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(315..320,321..338,339..345)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    353..369
                     /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     358..380
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     377..393
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     377..393
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(4608..4616,4617..4634,4635..4637,4638..4670,4671..4673,
                     4674..4697,4698..4724,4725..4781,4782..4787,4788..5057,
                     5058..5075,5076..5510,5511..5516,5517..5960)
                     /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      4704..4713
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     4994..5015
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5121..5142)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5382..5401)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(6879..6901)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7001..7020
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7098..7116)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7184..7288
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7289..7357,7358..8149)
                     /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(7507..7526)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8320..8908
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8809..8828
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
       61 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      121 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      181 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      241 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
      301 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      361 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct gtaagtttaa
      421 acatgatctt actaactaac tattctcatt taaattttca gagcttaaaa atggctgaaa
      481 tcactcacaa cgatggatac gctaacaact tggaaatgaa ataagcttgc atgcctgcag
      541 gtcgactcta gagttctcta aaggcactaa ccgtttaaca taaatgcaaa tcaattcaaa
      601 caaaaaccaa atgtttgtac tataggtttt tgaaaaaaaa cttgaaaatt ttagtcattg
      661 tacaaaaaac atcatgacat cagtattctg ctattcagct aagattcttg ttgtcgtcat
      721 ttgatgtatc cgttcgtcgg tgcaggactt ccacatggcg cagtcgaaac catgttaatc
      781 tctgaagtca aatggtctaa gaattcacag tatagacaga ctgcaggtgt aaacttatcc
      841 tatatatcac ccaagacgta ttaccttgtc agactgcatt tacacaagta atcccactta
      901 ttatatgatg tcacgcgtta taaaacggaa gcttacttat tgtggaacag gtggaatact
      961 caaaaacatt gaaaattctt acaatttcta agtaaaaacc ttatgaatgt tttgtacgta
     1021 ttattattcc cattcttcca tttagttgtc agtttttcat tgctcgggaa gatttgcata
     1081 aatatcgcca ttgaccagtc tactccccag ttttttgatc atgatttttc acaacaatga
     1141 tcaatcaaat aaacatcata tcatcttatc ccgattaagc gccctcgcaa ttttattgcc
     1201 tcatcttctt aataatagaa accatatgct catttttggc ttagtttttg ctcagttcaa
     1261 ttttccttat tccatcgtaa gcgttaagtc tagaaggctt tttgttattc actttttttc
     1321 tttgcttttt acttttctct ttcttcacga gacaaatatt ggtattattg ctcatctctc
     1381 tactaataaa actcaacaat ttcgttgaaa cttggtctct agagtttgat gagttgtttg
     1441 tgttattttt gatctaaatt ttcaattttg tggaagtaag gcttgcaaga tagacaaaga
     1501 caacaagaaa ataaacgaag ggattgagga agaataaact gaaatgatca tggttaagaa
     1561 aataggaaga aaatagaaga agagcggccg gctcactagg ttaaattatt tttgtgcttg
     1621 ttgtggggag cgttgagtaa gacctagagc aaggaacgcg gttagcaatc cctgccgttt
     1681 ccaaatctcc agagactatg cgaaattcag aagacacggg acatcaagac agataggtac
     1741 ggagtaagac cttcaaattg tttcttctct tcttgattac tctcttgctc ctctctcgtc
     1801 cattgtgctc tttcccacac actgagcaaa agcatcgtca ttgtggaaag tgagcttgat
     1861 gtggttggct caaacatctg accagctaga cttgtcactt gtaagatatc acatcgcact
     1921 tcccaattaa tcttaaaatc cgcttgactc atttattttt gaattcgttt ttgtttgaaa
     1981 ttgtaaactt cggaatagtt ccagaccgat tcagaactag tatgatcacc tatgaaacgt
     2041 gatttctgaa atattcttac ttttttctag acaactcaag atctcgatcg gccaaaattg
     2101 tgttatcctc gtaatcatca taaccatttc cgatattctg atcacaacgg ctttctttcg
     2161 aatctagtac gtctcacgta caactgccca caaacgattt ccatgaaact gataaataat
     2221 taaagacttt ggacttacca agagttcaaa caggttttat ttcgttttct atttttgaaa
     2281 aaaaaatcat tccagttgga cgacaaaaac agttgtagaa ctttaataat aatatttatt
     2341 cttgacacaa aaaaacttat cgtatttgta aggggaaaat agacccataa ttaacaacgt
     2401 ttctattagt tatgcatttt actgtactca tgatattact ccaataacaa ataatttatc
     2461 tgaaatagca cttttctcct tctccctgtg ctccaataca gtccagacac atattaattt
     2521 tcaacaccac atcacgcaca tcttccatag gaaattaaaa aaaaaataca ttagacttcc
     2581 tattcatatc aaatatttca catcataata tttttgtttg ctcatttttc aacacttttc
     2641 tgattcattt tatttttgta cgttacgata cctacatcag ttatttttca cctgccgtca
     2701 cttattttag atattcctta attttgacta cgtattactc attgttaaca tttctgatat
     2761 ttgtatactc aaagccattt ggaaagaatc agaacagtga tttcagtttt gatgctggcc
     2821 aactagttga ataacgtgat gaggaccatt gagtaattag agaaaacatg tttacgaata
     2881 tagtcggcta aattgagaaa aatacctaaa taactggcaa taagctcaga actgaaaaac
     2941 aaaaaattgc taaccatccc gtgaattttt gatgtgtgcc tgtctatgcc gtcatgttca
     3001 tctcatgtgc tttgttttca atccctggtc tcttgtaatt tttcttttaa aaagaagtct
     3061 gctattcgtg acaaattaaa attgaaaatt catatatctt tagagtatag ccgttttgag
     3121 tcagcaacaa acaattaggt gccacttccg ggacaggcac ctctgaaata aaaggaataa
     3181 gaaaatagtg agatattccc ccctcactga tcaaacttct ttttttccct ttactacgtt
     3241 tacctatatt tttattctgt tagtttggtc gaaaattggt ggaggatgtc tagaatttgc
     3301 aggatacaaa ttgagactaa agaaaattga atttcttcaa gtcttctcga tttcttgatt
     3361 tctagtaaag cttcctttgt aactactgta atagttagtg agaataatta aattttggcg
     3421 cctgccagtc atttctagtg aacaaatata acatcaatac tttgaaatct aattttctaa
     3481 gtagctcggg taattttttt gttcacatca ctccttacaa cacaactttc ttcaaaaatt
     3541 tcacgtctat atttagtttc aatttttcaa aggttgaagg cgtctagact tcgatgaata
     3601 ccaaaattaa agcaatcata gttttttagg taaagtttat gaatctgcaa agtcatacta
     3661 taaaactgat cttagaaatt tttcttgcag attttatctt aatatttcaa aaaacatcgt
     3721 tatgttccca ttttgatttt tggtgtaatg gtcgtgtttc tacgaacaaa aacggagaca
     3781 cccccaaaaa tttttgagat gctgtttttg aaaataataa aggcagctgt cagctttatt
     3841 catttttctg aaaaacaatt cactgtcatt ttgagctaat agctgcaaaa ccaacaaacg
     3901 taaaaactca gaaagcattc aaaaatcgta aaacttgagt ttttctcaag aaatacgatt
     3961 aagattactg tagacaaatt tatagtaaat tgaattaaag tgtaatttga taaatgcatg
     4021 actttctcag atttattcat tttccacata taaactctaa ttgaatacta ggaactgtgt
     4081 atgagtatat catttgttga aatgctacat ttttctgtta gtctagattt tttaacgagt
     4141 ttgtgtcttg gtaattgtcg tgtgtattca ttgttcacag cttctaatag gaaagggtta
     4201 ttttcaaaaa acgttctgta cttcgctggt aggagtactg tagttttact attttttttt
     4261 atataacaga actatttgtt tctgctttga aagcataatt cgaataatta cgtaggattc
     4321 aatttttctt gatactcatt ttcttttttg tctaaccatt tcattctgac acataaatta
     4381 ccaataacag attgattgtc tgtactttat gttattttcc tttcaatatt tttcctctct
     4441 cttatttcta aaagcggaaa tgaaaaacga ctctgagaat taattttctt catttcttct
     4501 gtaggcgtaa ctgcactttc tgggatttct ttttgctgtc ataatacggg agtatatttt
     4561 tttagaggaa tataaaactt ttaatttcta tttcagagga tccaaaaatg ggttctcatc
     4621 atcatcatca tcatggtatg gctagcatga ctggtggaca gcaaatgggt cgggatctgt
     4681 acgacgatga cgataaggat ctcgccacca tggtcgactc atcacgtcgt aagtggaata
     4741 agacaggtca cgcagtcaga gctataggtc ggctgagctc actcgagaac gtctatatca
     4801 aggccgacaa gcagaagaac ggcatcaagg cgaacttcca catccgccac aacatcgagg
     4861 acggcggcgt gcagctcgcc taccactacc agcagaacac ccccatcggc gacggccccg
     4921 tgctgctgcc cgacaaccac tacctgagcg tgcagtccaa actttcgaaa gaccccaacg
     4981 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     5041 tggacgagct gtacaagggc ggtaccggag ggagcatggt gagcaagggc gaggagctgt
     5101 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     5161 gcgtgtccgg cgagggtgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     5221 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     5281 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     5341 tgcccgaagg ctacatccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     5401 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     5461 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac ctgccggacc
     5521 aactgactga agagcagatc gcagaattta aagaggcttt ctccctattt gacaaggacg
     5581 gggatgggac aataacaacc aaggagctgg ggacggtgat gcggtctctg gggcagaacc
     5641 ccacagaagc agagctgcag gacatgatca atgaagtaga tgccgacggt gacggcacaa
     5701 tcgacttccc tgagttcctg acaatgatgg caagaaaaat gaaatacagg gacacggaag
     5761 aagaaattag agaagcgttc ggtgtgtttg ataaggatgg caatggctac atcagtgcag
     5821 cagagcttcg ccacgtgatg acaaaccttg gagagaagtt aacagatgaa gaggttgatg
     5881 aaatgatcag ggaagcagac atcgatgggg atggtcaggt aaactacgaa gagtttgtac
     5941 aaatgatgac agcgaagtaa gcggaattcc aactgagcgc cggtcgctac cattaccaac
     6001 ttgtctggtg tcaaaaataa taggggccgc tgtcatcaga gtaagtttaa actgagttct
     6061 actaactaac gagtaatatt taaattttca gcatctcgcg cccgtgcctc tgacttctaa
     6121 gtccaattac tcttcaacat ccctacatgc tctttctccc tgtgctccca ccccctattt
     6181 ttgttattat caaaaaactt ctcttaattt ctttgttttt tagcttcttt taagtcacct
     6241 ctaacaatga aattgtgtag attcaaaaat agaattaatt cgtaataaaa agtcgaaaaa
     6301 aattgtgctc cctcccccca ttaataataa ttctatccca aaatctacac aatgttctgt
     6361 gtacacttct tatgtttttt acttctgata aatttttttg aaacatcata gaaaaaaccg
     6421 cacacaaaat accttatcat atgttacgtt tcagtttatg accgcaattt ttatttcttc
     6481 gcacgtctgg gcctctcatg acgtcaaatc atgctcatcg tgaaaaagtt ttggagtatt
     6541 tttggaattt ttcaatcaag tgaaagttta tgaaattaat tttcctgctt ttgctttttg
     6601 gggtttcccc tattgtttgt caagatttcg aggacggcgt ttttcttgct aaaatcacaa
     6661 gtattgatga gcacgatgca agaaagatcg gaagaaggtt tgggtttgag gctcagtgga
     6721 aggtgagtag aagttgataa tttgaaagtg gagtagtgtc tatggggttt ttgccttaaa
     6781 tgacagaata cattcccaat ataccaaaca taactgtttc ctactagtcg gccgtacggg
     6841 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     6901 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     6961 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     7021 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     7081 atcaggcggc cttaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     7141 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     7201 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     7261 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     7321 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     7381 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     7441 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     7501 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     7561 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     7621 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     7681 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     7741 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     7801 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     7861 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     7921 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     7981 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     8041 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     8101 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     8161 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     8221 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     8281 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     8341 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     8401 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     8461 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     8521 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     8581 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     8641 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     8701 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     8761 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     8821 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     8881 ctcgtcaggg gggcggagcc tatggaaaaa cgccagc
//
