LOCUS       sequence-358855-        6831 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent expression of cell body-targeted GCaMP7f, under CAG 
            promoter. As bright as conventional GCaMP7f..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6831)
  AUTHORS   Shemesh OA, Linghu C, Piatkevich KD, Goodwin D, Celiker OT, Gritton 
            HJ, Romano MF, Gao R, Yu CJ, Tseng HA, Bensussen S, Narayan S, Yang 
            CT, Freifeld L, Siciliano CA, Gupta I, Wang J, Pak N, Yoon YG, 
            Ullmann JFP, Guner-Ataman B, Noamany H, Sheinkopf ZR, Park WM, Asano
            S, Keating AE, Trimmer JS, Reimer J, Tolias AS, Bear MF, Tye KM, Han
            X, Ahrens MB, Boyden ES
  TITLE     Precision Calcium Imaging of Dense Neural Populations via a 
            Cell-Body-Targeted Calcium Indicator.
  JOURNAL   Neuron. 2020 Jun 26. pii: S0896-6273(20)30398-6. doi: 
            10.1016/j.neuron.2020.05.029.
  PUBMED    32592656
REFERENCE   2  (bases 1 to 6831)
  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: "Neuron. 
            2020 Jun 26. pii: S0896-6273(20)30398-6. doi: 
            10.1016/j.neuron.2020.05.029."
COMMENT     Sequence Label: pAAV-CAG-FLEX-SomaGCaMP7
FEATURES             Location/Qualifiers
     source          1..6831
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2069..2090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2281..2300
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2459..2588
                     /label=AAV2 ITR
     primer_bind     3467..3489
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3551..3570
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    3605..3638
                     /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    3701..3734
                     /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(4099..4539,4540..4545,4546..4980,4981..4998,
                     4999..5268,5269..5271,5272..5274,5275..5331,5332..5358,
                     5359..5382,5383..5385,5386..5418,5419..5421,5422..5439,
                     5440..5448))
                     /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     4655..4674
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4914..4935
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5041..5062)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      5343..5352
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      5445..5454
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(5471..5504)
                     /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(5575..5608)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5641..6229
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5694..5714)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6112..6123)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6270..6477
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6647..6776
                     /label=AAV2 ITR
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag
     2641 gaagatcgta ggttaattaa ccattgacgt caataatgac gtatgttccc atagtaacgc
     2701 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     2761 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     2821 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     2881 tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc
     2941 tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt
     3001 gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc gggggggggg gggggggggg
     3061 gggggggggg sggggsgrgg sggrgaggtg cggcggcagc caatcagagc ggcgcgctcc
     3121 gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc
     3181 ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc
     3241 gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc
     3301 cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc
     3361 tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggc
     3421 tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg
     3481 cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta
     3541 cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattgaat
     3601 tcccataact tcgtataaag tatcctatac gaagttatat caaaatagga agaccaatgc
     3661 ttcaccatcg acccgaattg ccaagcatca ccatcgaccc ataacttcgt ataatgtatg
     3721 ctatacgaag ttatactagc tagcttacag aggtccgtag cgtgtctcgt actggctcac
     3781 gatgttgcgc agccgctgca ctctctgcct cagctcagcc acccttgtcc tcagggcggt
     3841 gttccgtctc ctcagaaaag cggcccggat ctccagggat cccagtggtc cgtatctggt
     3901 ctcgtactga gacacgatgt tctccagccg ctgcacctcc tgctccagct cagccacctc
     3961 tgtctccagg gcggtgttct cctgctccag gaaagcggcc tcgatttcca gtgtcccacc
     4021 gctaccgccg cttccccctg tcccaccgct accaccgcta cccccagtcc ctccagaccc
     4081 accgctgccc ccgagctcct tcgctgtcat catttgtaca aactcttcgt agtttacctg
     4141 accatcccca tcgatgtctg cttccctgat catttcatca acctcttcat ctgttaactt
     4201 ctctccaagg ttgttcatca cgtggcgaag ctctgctgca ctgatgtagc cattgccatc
     4261 cttatcaaac acaccgaacg cttctctaat ttcttcttca ctgtccctgt agctcccttt
     4321 tcttgccatc attgtcagga actcagggaa gtcgattgtg ccgtcaccgt cggcatctac
     4381 ttcattgatc atgtcctgca gctctgcttc tgtggggttc tgccccagag accgcatcac
     4441 cgtccccagc tccttggttg ttattgtccc atccccgtcc ttgtcaaata gggagaaagc
     4501 ctctttaaat tctgcgatct gctcttcagt cagttggtcc ggcaggttgt actccagctt
     4561 gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac
     4621 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa
     4681 gaagatggtg cgctcctgga tgtagccttc gggcatggcg gacttgaaga agtcgtgctg
     4741 cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg tggtcacgag
     4801 ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc
     4861 gtaggtggca tcgccctcac cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc
     4921 gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat
     4981 gctccctccg gttccgccct tgtacagctc gtccatgccg agagtgatcc cggcggcggt
     5041 cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttcg aaagtttgga
     5101 ctgcacgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt
     5161 ctgctggtag tggtaggcga gctgcacgcc gccgtcctcg atgttgtggc ggatcttgaa
     5221 gttcgccttg atgccgttct tctgcttgtc ggccttgata tagacgttag gctgtgaact
     5281 cagccgacct atagctctga ctgcgtgacc tgtcttattc cacttacgac gtgatgagtc
     5341 gaccatggtg gcgagatcct tatcgtcatc gtcgtacaga tcccgaccca tttgctgtcc
     5401 accagtcatg gaagccatac catgatgatg atgatgatga gaacccatgg tggcactagt
     5461 ggtacctagt ataacttcgt ataggatact ttatacgaag ttatcattgg gattcttcct
     5521 attttgatcc aagcatcacc atcgaccctc tagtccagat ctcaccatcg acccataact
     5581 tcgtatagca tacattatac gaagttatgt ccctcgaaga ggttcgcgaa gcttatcgat
     5641 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     5701 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     5761 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     5821 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     5881 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     5941 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     6001 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     6061 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     6121 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     6181 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt
     6241 cgacccgggc ggccgctcat aaagctcgac tgtgccttct agttgccagc catctgttgt
     6301 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     6361 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     6421 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     6481 ggtccggact gtacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg
     6541 ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat cttcctagag
     6601 catggctacg tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg
     6661 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     6721 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     6781 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc t
//
