LOCUS       sequence-12266-e        5847 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5847)
  AUTHORS   Dreosti E, Odermatt B, Dorostkar MM, Lagnado L
  TITLE     A genetically encoded reporter of synaptic activity in vivo.
  JOURNAL   Nat Methods. 2009 Dec . 6(12):883-9.
  PUBMED    19898484
REFERENCE   2  (bases 1 to 5847)
  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: "Nat 
            Methods. 2009 Dec . 6(12):883-9."
COMMENT     Sequence Label: I-SceI pB 5UAS::ZfSyGCaMP2
FEATURES             Location/Qualifiers
     source          1..5847
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    671..765
                     /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)"
     promoter        784..1022
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     969..990
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     regulatory      1101..1110
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2013..2024,2025..2042,2043..2045,2046..2078,2079..2081,
                     2082..2105,2106..2132,2133..2189,2190..2195,2196..2465,
                     2466..2483,2484..2918,2919..2921,2922..2924,2925..3368)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MRGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWN
                     KTGHAVRAIGRLSSLENVYIMADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDG
                     PVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGE
                     ELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTL
                     TYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRI
                     ELKGIDFKEDGNILGHKLEYNTRDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMR
                     SLGQNPTEAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDG
                     NGYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             join(2013..2024,2025..2042,2043..2045,2046..2078,2079..2081,
                     2082..2105,2106..2132,2133..2189,2190..2195,2196..2465,
                     2466..2483,2484..2918,2919..2921,2922..2924,2925..3368)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MRGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWN
                     KTGHAVRAIGRLSSLENVYIMADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDG
                     PVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGE
                     ELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTL
                     TYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRI
                     ELKGIDFKEDGNILGHKLEYNTRDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMR
                     SLGQNPTEAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDG
                     NGYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      2112..2121
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2402..2423
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2529..2550)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2790..2809)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    3490..3611
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3527..3546)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3581..3600
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3624..3640)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3625..3641)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3698..3714)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3698..3714)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3711..3733)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3722..3738
                     /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)."
     promoter        complement(join(3746..3752,3753..3770,3771..3776))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3791..3812
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3929..3946)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4100..4688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4180..4199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4859..5650,5651..5719))
                     /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     5482..5501
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5720..5824)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcattacc ctgttatccc taaagggaac aaaagctgga
      661 gcttaatggt cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc
      721 ctccgagcgg agtactgtcc tccgagcgga gtactgtcct ccgagcggag actctagcga
      781 gcgccggagt ataaatagag gcgcttcgtc tacggagcga caattcaatt caaacaagca
      841 aagtgaacac gtcgctaagc gaaagctaag caaataaaca agcgcagctg aacaagctaa
      901 acaatctgca gtaaagtgca agttaaagtg aatcaattaa aagtaaccag caaccaagta
      961 aatcaactgc aactactgaa atctgccaag aagtaattat tgaatacaag aagagaactc
     1021 tgaataggga attggctagc gctaccggac tcagatcccc cgggctgcag gaattcgatg
     1081 ctaccggact cagatctcga gccaccatgg atgttgccaa ccagttggtc gccactgggc
     1141 agtttacaat cgtcaaacag ccattaggat tcatcaaaat cttacaatgg ttctttgcca
     1201 tcttcgcgtt cgccacctgc ggtggttact ctggagtgtt tcgtatgagc gtggagtgtg
     1261 agaaccgctc agagagtgac ctgaacattg aggttgagtt tgcgtacccg ttcagactgc
     1321 accaggtatg gtttgatgcc cctacgtgta aaggatcaga acccgagcga ctctttctgg
     1381 tgggagacaa ctcttcctca gcagagttct tcgtcaccat cggcgtcttc tccttccttt
     1441 attcaatggc agccatttct gtgtacatct tcctgttgga gaaataccgc gagggaaaca
     1501 aaggagctca ggctgatttc attgtgacag cggttttcac cttcatgtgg ctggtgagtt
     1561 catcggcgtg ggctaaaggt ctctcagacg tgaagagagc caccgaccct gatgacgtca
     1621 tcaacctcat ccctgcgtgc gaacgagagg aaaaccgatg caaagagctg cacgaaccgg
     1681 tggtgtctgg cctgaacaca tctgtggcat tcgggttctg taatgtgatc ctatggggag
     1741 gcaacctctg gttcatcttc aaggaaaccg gctggatggc ggctttcgct ggcacctaca
     1801 tgccatctgg agagaagcag ccagctccag actccttcgg ccagggctat ggacaggaag
     1861 gttatggaca ggaccagtat gccagctctc agggtggata tcagcctgac tacggccagc
     1921 agggtggata tgaaggcggt ggatacaatc aaggagtgta tggtcaggga gagcccacat
     1981 ccttctccaa cgaggatcca ccggtcgcca ccatgcgggg ttctcatcat catcatcatc
     2041 atggtatggc tagcatgact ggtggacagc aaatgggtcg ggatctgtac gacgatgacg
     2101 ataaggatct cgccaccatg gtcgactcat cacgtcgtaa gtggaataag acaggtcacg
     2161 cagtcagagc tataggtcgg ctgagctcac tcgagaacgt ctatatcatg gccgacaagc
     2221 agaagaacgg catcaaggcg aacttcaaga tccgccacaa catcgaggac ggcggcgtgc
     2281 agctcgccta ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     2341 acaaccacta cctgagcacc cagtccaaac tttcgaaaga ccccaacgag aagcgcgatc
     2401 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     2461 acaagggcgg taccggaggg agcatggtga gcaagggcga ggagctgttc accggggtgg
     2521 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     2581 agggtgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     2641 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     2701 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2761 acatccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     2821 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2881 aggacggcaa catcctgggg cacaagctgg agtacaacac gcgtgaccaa ctgactgaag
     2941 agcagatcgc agaatttaaa gaggctttct ccctatttga caaggacggg gatgggacaa
     3001 taacaaccaa ggagctgggg acggtgatgc ggtctctggg gcagaacccc acagaagcag
     3061 agctgcagga catgatcaat gaagtagatg ccgacggtaa tggcacaatc gacttccctg
     3121 agttcctgac aatgatggca agaaaaatga aagacacaga cagtgaagaa gaaattagag
     3181 aagcgttccg tgtgtttgat aaggatggca atggctacat cagtgcagca gagcttcgcc
     3241 acgtgatgac aaaccttgga gagaagttaa cagatgaaga ggttgatgaa atgatcaggg
     3301 aagcagacat cgatggggat ggtcaggtaa actacgaaga gtttgtacaa atgatgacag
     3361 cgaagtgagc ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta
     3421 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     3481 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3541 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3601 aatgtatctt aaatcaagct tatcgatacc gtcgacctcg agggggggcc cggtacccaa
     3661 ttaccctgtt atccctagcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     3721 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     3781 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     3841 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3901 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3961 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4021 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4081 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4141 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4201 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4261 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4321 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4381 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4441 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4501 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     4561 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4621 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4681 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4741 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4801 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4861 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4921 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4981 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5041 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5101 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5161 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5221 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5281 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5341 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5401 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5461 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5521 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5581 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5641 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5701 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5761 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5821 cgcgcacatt tccccgaaaa gtgccac
//
