LOCUS       sequence-432424-        6178 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector with hSynapsin promoter and DIO Lox sites for Cre-On 
            expression of green fluorescent calcium indicator GCaMP6f.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6178)
  AUTHORS   Meng C, Zhou J, Papaneri A, Peddada T, Xu K, Cui G
  TITLE     Spectrally Resolved Fiber Photometry for Multi-component Analysis of
            Brain Circuits.
  JOURNAL   Neuron. 2018 May 16;98(4):707-717.e4. doi: 
            10.1016/j.neuron.2018.04.012. Epub 2018 May 3.
  PUBMED    29731250
REFERENCE   2  (bases 1 to 6178)
  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: 
            "10.1016/j.neuron.2018.04.012"; journalName: "Neuron"; date: 
            "2018-05-16- 16"; volume: "98"; issue: "4"; pages: "707-717.e4"
COMMENT     Sequence Label: pAAV-hSyn-DIO-GCaMP6f-WPRE
FEATURES             Location/Qualifiers
     source          1..6178
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(660..693)
                     /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(744..777)
                     /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(796..1239,1240..1245,1246..1680,1681..1698,
                     1699..1968,1969..1974,1975..2031,2032..2058,2059..2082,
                     2083..2085,2086..2118,2119..2121,2122..2139,2140..2148))
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     primer_bind     1355..1374
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1614..1635
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1741..1762)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      2043..2052
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2145..2154
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2163..2196
                     /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    2247..2280
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2305..2893
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2358..2378)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2776..2787)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2925..3401
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3124..3143)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3441..3581
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3441..3570
                     /label=AAV2 ITR
     rep_origin      3656..4111
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3743..3762)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3953..3974
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4128..4147)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4247..4269
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4307..4325)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4393..4497
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4498..4566,4567..5358)
                     /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(4716..4735)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5529..6117
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6018..6037
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gactccggaa
      661 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
      721 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
      781 gcgagtcgcg gccgctcact tcgctgtcat catttgtaca aactcttcgt agtttacctg
      841 accatcccca tcgatgtctg cttccctgat catttcatca acctcttcat ctgttaactt
      901 ctctccaagg tttgtcatca cgtggcgaag ctctgctgca ctgatgtagc cattgccatc
      961 cttatcaaac acaccgaacg cttctctaat ttcttcttcc gtgtccctgt atttcatttt
     1021 tcttgccatc attgtcagga actcagggaa gtcgattgtg ccgtcaccgt cggcatctac
     1081 ttcattgatc atgtcctgca gctctgcttc tgtggggttc tgccccagag accgcatcac
     1141 cgtccccagc tccttggttg ttattgtccc atccccgtcc ttgtcaaata gggagaattc
     1201 ctctttaaat tctgcgatct gctcttcagt cagttggtcc ggcaggttgt actccagctt
     1261 gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac
     1321 cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa
     1381 gaagatggtg cgctcctgga tgtagccttc gggcatggcg gacttgaaga agtcgtgctg
     1441 cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg tggtcacgag
     1501 ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc
     1561 gtaggtggca tcgccctcac cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc
     1621 gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat
     1681 gctccctccg gtaccgccct tgtacagctc gtccatgccg agagtgatcc cggcggcggt
     1741 cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttcg aaagtttgga
     1801 ctgcacgctc aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt
     1861 ctgctggtag tggtaggcga gctgcacgcc gccgtcctcg atgttgtggc ggatcttgaa
     1921 gttcgccttg atgccgttct tctgcttgtc ggccttgata tagacgttct cgagtgagct
     1981 cagccgacct atagctctga ctgcgtgacc tgtcttattc cacttacgac gtgatgagtc
     2041 gaccatggtg gcgagatcct tatcgtcatc gtcgtacaga tcccgaccca tttgctgtcc
     2101 accagtcatg ctagccatac catgatgatg atgatgatga gaacccatgg tggcgagcta
     2161 gcataacttc gtataaagta tcctatacga agttatttgc cttaacccag aaattatcac
     2221 tgttattctt tagaatggtg caaagaataa cttcgtataa tgtatgctat acgaagttat
     2281 gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     2341 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     2401 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     2461 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     2521 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     2581 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     2641 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     2701 tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc
     2761 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     2821 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     2881 gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg
     2941 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg
     3001 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg
     3061 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg
     3121 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg
     3181 cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat
     3241 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg
     3301 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc
     3361 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca
     3421 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     3481 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     3541 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     3601 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     3661 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3721 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3781 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3841 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     3901 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3961 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     4021 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4081 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     4141 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4201 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4261 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     4321 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4381 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4441 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4501 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4561 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4621 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4681 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4741 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4801 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4861 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4921 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4981 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5041 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5101 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5161 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5221 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5281 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5341 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5401 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5461 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5521 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5581 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5641 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5701 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5761 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5821 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5881 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5941 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6001 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6061 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6121 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
