LOCUS       sequence-431696-        5996 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector with hSynapsin promoter and LoxFAS sites for Cre-Off 
            expression of GCaMP6f (GCaMP6f will not be expressed in neurons that
            express Cre recombinase).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5996)
  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 5996)
  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-FAS-GCaMP6f-WPRE
FEATURES             Location/Qualifiers
     source          1..5996
                     /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)"
     CDS             join(702..710,711..728,729..731,732..764,765..767,768..791,
                     792..818,819..875,876..881,882..1151,1152..1169,1170..1604,
                     1605..1610,1611..2051)
                     /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"
     regulatory      798..807
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1088..1109
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1215..1236)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1476..1495)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2123..2711
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2176..2196)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2594..2605)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2743..3219
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2942..2961)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3259..3399
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3259..3388
                     /label=AAV2 ITR
     rep_origin      3474..3929
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3561..3580)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3771..3792
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3946..3965)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4065..4087
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4125..4143)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4211..4315
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4316..4384,4385..5176)
                     /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(4534..4553)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5347..5935
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5836..5855
                     /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 caacttcgta tatacctttc tatacgaagt tgtggcgcgc catgggttct catcatcatc
      721 atcatcatgg tatggctagc atgactggtg gacagcaaat gggtcgggat ctgtacgacg
      781 atgacgataa ggatctcgcc accatggtcg actcatcacg tcgtaagtgg aataagacag
      841 gtcacgcagt cagagctata ggtcggctga gctcactcga gaacgtctat atcaaggccg
      901 acaagcagaa gaacggcatc aaggcgaact tcaagatccg ccacaacatc gaggacggcg
      961 gcgtgcagct cgcctaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1021 tgcccgacaa ccactacctg agcgtgcagt ccaaactttc gaaagacccc aacgagaagc
     1081 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     1141 agctgtacaa gggcggtacc ggagggagca tggtgagcaa gggcgaggag ctgttcaccg
     1201 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     1261 ccggcgaggg tgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     1321 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     1381 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     1441 aaggctacat ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     1501 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1561 tcaaggagga cggcaacatc ctggggcaca agctggagta caacctgccg gaccaactga
     1621 ctgaagagca gatcgcagaa tttaaagagg aattctccct atttgacaag gacggggatg
     1681 ggacaataac aaccaaggag ctggggacgg tgatgcggtc tctggggcag aaccccacag
     1741 aagcagagct gcaggacatg atcaatgaag tagatgccga cggtgacggc acaatcgact
     1801 tccctgagtt cctgacaatg atggcaagaa aaatgaaata cagggacacg gaagaagaaa
     1861 ttagagaagc gttcggtgtg tttgataagg atggcaatgg ctacatcagt gcagcagagc
     1921 ttcgccacgt gatgacaaac cttggagaga agttaacaga tgaagaggtt gatgaaatga
     1981 tcagggaagc agacatcgat ggggatggtc aggtaaacta cgaagagttt gtacaaatga
     2041 tgacagcgaa ggataacggc tagcacaact tcgtatatac ctttctatac gaagttgtga
     2101 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     2161 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2221 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2281 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     2341 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     2401 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     2461 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     2521 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt
     2581 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     2641 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     2701 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac
     2761 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     2821 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     2881 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     2941 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     3001 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     3061 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     3121 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     3181 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     3241 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     3301 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     3361 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     3421 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     3481 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3541 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3601 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3661 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     3721 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3781 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     3841 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3901 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     3961 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     4021 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     4081 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     4141 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4201 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4261 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4321 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4381 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4441 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4501 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4561 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4621 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4681 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4741 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4801 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4861 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     4921 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     4981 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5041 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5101 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5161 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5221 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5281 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5341 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5401 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5461 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5521 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5581 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5641 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5701 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5761 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5821 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5881 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5941 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
