LOCUS       sequence-229564-        5891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For Calcium imaging, expression of G-CaMP3 under the expression of a
            Pinx-1 promoter (AIB interneurons).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5891)
  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 5891)
  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_Pinx-1::G-CaMP3
FEATURES             Location/Qualifiers
     source          1..5891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     114..133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     367..384
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    501..522
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(537..542,543..560,561..567)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    575..591
                     /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     580..602
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     599..615
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     599..615
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(1804..1812,1813..1830,1831..1833,1834..1866,1867..1869,
                     1870..1893,1894..1920,1921..1977,1978..1983,1984..2253,
                     2254..2271,2272..2706,2707..2709,2710..2712,2713..3156)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNTRDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             join(1804..1812,1813..1830,1831..1833,1834..1866,1867..1869,
                     1870..1893,1894..1920,1921..1977,1978..1983,1984..2253,
                     2254..2271,2272..2706,2707..2709,2710..2712,2713..3156)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNTRDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      1900..1909
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2190..2211
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2317..2338)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2578..2597)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(4075..4097)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4197..4216
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4294..4312)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4380..4484
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4485..4553,4554..5345)
                     /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(4703..4722)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5516..5891,1..213)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
       61 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
      121 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
      181 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
      241 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
      301 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
      361 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
      421 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
      481 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
      541 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
      601 ggaaacagct atgaccatga ttacgccaag ctgtaagttt aaacatgatc ttactaacta
      661 actattctca tttaaatttt cagagcttaa aaatggctga aatcactcac aacgatggat
      721 acgctaacaa cttggaaatg aaataagctt gcatgcctgc aggtcgactc tagagttact
      781 aatagattca aaaattattt tctgtgcttt tcacaaatac actttatttt aggtaatcat
      841 attttaatta aacacgcggg aaatttttta gggtaaactt tgaagttttt tttgtcgcgg
      901 atatcagatt cataactgtc tatgtaccat aaattacgac ttttgaatat ttgctgttca
      961 aagtcaatgt ataaattaaa tttaagtttc tttcgtcata catctcaaaa atagctaaaa
     1021 tgcgtttaac aaatgttata cctaattttc aatcataact tggagagtat gaatgaattc
     1081 aaacgtttcc ttgaggtgat atccttttgg tacttgtata ttatcgaggc acagtttgaa
     1141 aataaattaa attttaattc atttgatgat tttgttttct cttgaggctt aaaaatgata
     1201 aacggtacaa aactacaaaa aaactccata agtctttatt ttcttaattt ttgaaatttt
     1261 atttcaaaat gcacaagatc catttatcat atttatagtt cttcatcttc ttttttttaa
     1321 tatcctcttt ttgtgtttta ttcggtccat acgtgctctt caactttttt ttcatttttg
     1381 actttaaata agaaaagatg gcaataaaaa atttggccga agagggcgat ggacggatga
     1441 aaatctacta aaaggattat aactcaattt gatatgctct tcgggaggta tcctcctgac
     1501 gagatggaaa agaagaagaa gaagaagaag aagcttgtca tcgtttcatc ggcaaagaga
     1561 cgggtggaca ttagaccacg ccccacaggg aaacctttcg agtttcatcc acctctgtct
     1621 gtttaacata tttgtttttt tctagctcta tttttcttcc gcttctactg cgtacttttg
     1681 cataattcta tttctaagtc tgatcattat aagtatcatc ctgaacatcg cacactaaac
     1741 atcctcggcc ggcacggatg aagtagtttt cattgcagtt cttgtccgcc ggaggatcca
     1801 aaaatgggtt ctcatcatca tcatcatcat ggtatggcta gcatgactgg tggacagcaa
     1861 atgggtcggg atctgtacga cgatgacgat aaggatctcg ccaccatggt cgactcatca
     1921 cgtcgtaagt ggaataagac aggtcacgca gtcagagcta taggtcggct gagctcactc
     1981 gagaacgtct atatcaaggc cgacaagcag aagaacggca tcaaggcgaa cttcaagatc
     2041 cgccacaaca tcgaggacgg cggcgtgcag ctcgcctacc actaccagca gaacaccccc
     2101 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcgtgca gtccaaactt
     2161 tcgaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2221 gggatcactc tcggcatgga cgagctgtac aagggcggta ccggagggag catggtgagc
     2281 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     2341 aacggccaca agttcagcgt gtccggcgag ggtgagggcg atgccaccta cggcaagctg
     2401 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     2461 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     2521 ttcttcaagt ccgccatgcc cgaaggctac atccaggagc gcaccatctt cttcaaggac
     2581 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     2641 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     2701 tacaacacgc gtgaccaact gactgaagag cagatcgcag aatttaaaga ggctttctcc
     2761 ctatttgaca aggacgggga tgggacaata acaaccaagg agctggggac ggtgatgcgg
     2821 tctctggggc agaaccccac agaagcagag ctgcaggaca tgatcaatga agtagatgcc
     2881 gacggtgacg gcacaatcga cttccctgag ttcctgacaa tgatggcaag aaaaatgaaa
     2941 gacacagaca gtgaagaaga aattagagaa gcgttccgtg tgtttgataa ggatggcaat
     3001 ggctacatca gtgcagcaga gcttcgccac gtgatgacaa accttggaga gaagttaaca
     3061 gatgaagagg ttgatgaaat gatcagggaa gcagacatcg atggggatgg tcaggtaaac
     3121 tacgaagagt ttgtacaaat gatgacagcg aagtaagcgg aattccaact gagcgccggt
     3181 cgctaccatt accaacttgt ctggtgtcaa aaataatagg ggccgctgtc atcagagtaa
     3241 gtttaaactg agttctacta actaacgagt aatatttaaa ttttcagcat ctcgcgcccg
     3301 tgcctctgac ttctaagtcc aattactctt caacatccct acatgctctt tctccctgtg
     3361 ctcccacccc ctatttttgt tattatcaaa aaacttctct taatttcttt gttttttagc
     3421 ttcttttaag tcacctctaa caatgaaatt gtgtagattc aaaaatagaa ttaattcgta
     3481 ataaaaagtc gaaaaaaatt gtgctccctc cccccattaa taataattct atcccaaaat
     3541 ctacacaatg ttctgtgtac acttcttatg ttttttactt ctgataaatt tttttgaaac
     3601 atcatagaaa aaaccgcaca caaaatacct tatcatatgt tacgtttcag tttatgaccg
     3661 caatttttat ttcttcgcac gtctgggcct ctcatgacgt caaatcatgc tcatcgtgaa
     3721 aaagttttgg agtatttttg gaatttttca atcaagtgaa agtttatgaa attaattttc
     3781 ctgcttttgc tttttggggt ttcccctatt gtttgtcaag atttcgagga cggcgttttt
     3841 cttgctaaaa tcacaagtat tgatgagcac gatgcaagaa agatcggaag aaggtttggg
     3901 tttgaggctc agtggaaggt gagtagaagt tgataatttg aaagtggagt agtgtctatg
     3961 gggtttttgc cttaaatgac agaatacatt cccaatatac caaacataac tgtttcctac
     4021 tagtcggccg tacgggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     4081 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     4141 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     4201 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     4261 aaggagaaaa taccgcatca ggcggcctta agggcctcgt gatacgccta tttttatagg
     4321 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     4381 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4441 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4501 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4561 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4621 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4681 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     4741 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4801 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4861 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     4921 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     4981 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5041 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5101 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5161 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5221 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5281 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5341 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5401 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5461 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5521 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5581 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5641 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5701 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5761 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5821 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5881 ccgaactgag a
//
