LOCUS       sequence-213536-        5933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of ultrasensitive protein calcium sensor 
            under the Syn promoter, Cre-dependent expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5933)
  AUTHORS   Dana H, Sun Y, Mohar B, Hulse BK, Kerlin AM, Hasseman JP, Tsegaye G,
            Tsang A, Wong A, Patel R, Macklin JJ, Chen Y, Konnerth A, Jayaraman 
            V, Looger LL, Schreiter ER, Svoboda K, Kim DS
  TITLE     High-performance calcium sensors for imaging activity in neuronal 
            populations and microcompartments.
  JOURNAL   Nat Methods. 2019 Jul;16(7):649-657. doi: 10.1038/s41592-019-0435-6.
            Epub 2019 Jun 17.
  PUBMED    31209382
REFERENCE   2  (bases 1 to 5933)
  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"; doi: 
            "10.1038/s41592-019-0435-6"; journalName: "Nat Methods"; date: 
            "2019-07"; volume: "16"; issue: "7"; pages: "649-657"
COMMENT     Sequence Label: pGP-AAV-syn-FLEX-jGCaMP7c variant 1513-WPRE
FEATURES             Location/Qualifiers
     source          1..5933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   45..174
                     /label=AAV2 ITR
     promoter        256..703
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    733..766
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    829..862
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(873..1316,1317..1322,1323..1757,1758..1775,
                     1776..2045,2046..2048,2049..2051,2052..2108,2109..2135,
                     2136..2159,2160..2162,2163..2195,2196..2198,2199..2216,
                     2217..2225))
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6m
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSQPNVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKGSYRDSEEEIREAFGVFDKDGN
                     GYISAAELRHVMNNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     primer_bind     1432..1451
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1691..1712
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1818..1839)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      2120..2129
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2222..2231
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2242..2275)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(2346..2379)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2421..3009
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2474..2494)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2892..2903)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3052..3173
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3063..3082)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3117..3136
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3335..3464
                     /label=AAV2 ITR
     rep_origin      complement(3701..4289)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3781..3800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4460..5251,5252..5320))
                     /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     5083..5102
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5321..5425)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5451..5906)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5588..5609)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5800..5819
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctgcgc gctcgctcgc
       61 tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag
      121 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt
      181 taatgattaa cccgccatgc tacttatcta cgtagccatg ctctaggaag atctctgcag
      241 agggccctgc gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc
      301 ctacctgacg accgaccccg acccactgga caagcaccca acccccattc cccaaattgc
      361 gcatccccta tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc
      421 agcttcagca ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct
      481 cagcactgaa ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca
      541 ccttggtcgc gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat
      601 aggggggcac gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc
      661 tgcggtgggc agcggaggag tcgtgtcgtg cctgagagcg cagtcgaatt caagctgcta
      721 gcaaggatcc ccataacttc gtataaagta tcctatacga agttatatca aaataggaag
      781 accaatgctt caccatcgac ccgaattgcc aagcatcacc atcgacccat aacttcgtat
      841 aatgtatgct atacgaagtt atactagagc tttcacttcg ctgtcatcat ttgtacaaac
      901 tcttcgtagt ttacctgacc atccccatcg atgtctgctt ccctgatcat ttcatcaacc
      961 tcttcatctg ttaacttctc tccaaggttg ttcatcacgt ggcgaagctc tgctgcactg
     1021 atgtagccat tgccatcctt atcaaacaca ccgaacgctt ctctaatttc ttcttcactg
     1081 tccctgtagc tcccttttct tgccatcatt gtcaggaact cagggaagtc gattgtgccg
     1141 tcaccgtcgg catctacttc attgatcatg tcctgcagct ctgcttctgt ggggttctgc
     1201 cccagagaac gcatcaccgt ccccagctcc ttggttgtta ttgtcccatc cccgtccttg
     1261 tcaaataggg agaaagcctc tttaaattct gcgatctgct cttcagtcag ttggtcagga
     1321 aggttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc
     1381 agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag
     1441 ttgccgtcgt ccttgaagaa gatggtgcgc tcctggatgt agccttcggg catggcggac
     1501 ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc tgaagcactg cacgccgtag
     1561 gtcagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt gcagatgaac
     1621 ttcagggtca gcttgccgta ggtggcatcg ccctcaccct cgccggacac gctgaacttg
     1681 tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc
     1741 tcgcccttgc tcaccatgct ccctccggta ccgcccttgt acagctcgtc catgccgaga
     1801 gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg
     1861 tctttcgaaa gtttggactg cacgctcagg tagtggttgt cgggcagcag cacggggccg
     1921 tcgccgatgg gggtgttctg ctggtagtgg taggcgagct gcacgccgcc gtcctcgatg
     1981 ttgtggcgga tcttgaagtt cgccttgatg ccgttcttct gcttgtcggc cttgatatag
     2041 acgttaggct gtgagctcag ccgacctata gctctgactg cgtgacctgt cttattccac
     2101 ttacgacgtg atgagtcgac catggtggcg agatccttat cgtcatcgtc gtacagatcc
     2161 cgacccattt gctgtccacc agtcatgcta gccataccat gatgatgatg atgatgagaa
     2221 cccatggtgg cggatcctag tataacttcg tataggatac tttatacgaa gttatcattg
     2281 ggattcttcc tattttgatc caagcatcac catcgaccct ctagtccaga tctcaccatc
     2341 gacccataac ttcgtatagc atacattata cgaagttatg tccctcgaag aggttcgaat
     2401 tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa agattgactg
     2461 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     2521 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     2581 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     2641 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     2701 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     2761 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     2821 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     2881 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     2941 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     3001 cctccccgca tcgataccgt cgacccgggc ggccgcttcg agcagacatg ataagataca
     3061 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     3121 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     3181 acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagca
     3241 agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct tcctagagca tggctacgta
     3301 gataagtagc atggcgggtt aatcattaac tacaaggaac ccctagtgat ggagttggcc
     3361 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     3421 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcat taatgaatcg
     3481 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     3541 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3601 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3661 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3721 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3781 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3841 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3901 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3961 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4021 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4081 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4141 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4201 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4261 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4321 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4381 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4441 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4501 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4561 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4621 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4681 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4741 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4801 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4861 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4921 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4981 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5041 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5101 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5161 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5221 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5281 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5341 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5401 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag
     5461 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     5521 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     5581 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     5641 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     5701 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     5761 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     5821 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     5881 gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca act
//
