LOCUS       sequence-296254-        6926 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cre-conditional expression of synaptoPAC, a fusion of synaptophysin,
            mScarlet, and bPAC in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6926)
  AUTHORS   Oldani S, Moreno-Velasquez L, Faiss L, Stumpf A, Rosenmund C, 
            Schmitz D, Rost BR
  TITLE     SynaptoPAC, an optogenetic tool for induction of presynaptic 
            plasticity.
  JOURNAL   J Neurochem. 2021 Feb;156(3):324-336. doi: 10.1111/jnc.15210. Epub 
            2020 Oct 22.
  PUBMED    33037623
REFERENCE   2  (bases 1 to 6926)
  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: "10.1111/jnc.15210";
            journalName: "J Neurochem"; date: "2021-02"; volume: "156"; issue: 
            "3"; pages: "324-336"
COMMENT     Sequence Label: pAAV-syn-FLEX-synaptoPAC-minWPRE
FEATURES             Location/Qualifiers
     source          1..6926
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     repeat_region   20..160
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   31..160
                     /label=AAV2 ITR
     promoter        192..639
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(654..687)
                     /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(738..771)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(827..844)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(854..883)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(1952..2647)
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(2227..2246)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2496..2514
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    3633..3666
                     /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    3717..3750
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(3812..3832)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4208..4348
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4208..4337
                     /label=AAV2 ITR
     rep_origin      4423..4878
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4510..4529)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4720..4741
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4895..4914)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5014..5036
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5074..5092)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5160..5264
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5265..5333,5334..6125)
                     /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(5483..5502)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6296..6884
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6785..6804
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
       61 cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg
      121 cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgcac gcgtagaggg
      181 ccctgcgtat gagtgcaagt gggttttagg accaggatga ggcggggtgg gggtgcctac
      241 ctgacgaccg accccgaccc actggacaag cacccaaccc ccattcccca aattgcgcat
      301 cccctatcag agagggggag gggaaacagg atgcggcgag gcgcgtgcgc actgccagct
      361 tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc
      421 actgaaggcg cgctgacgtc actcgccggt cccccgcaaa ctccccttcc cggccacctt
      481 ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg cgagataggg
      541 gggcacgggc gcgaccatct gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg
      601 gtgggcagcg gaggagtcgt gtcgtgcctg agagcgcagt cgctagtacc ggtataactt
      661 cgtataggat actttatacg aagttatgca gaatggtagc tggattgtag ctgctattag
      721 caatatgaaa cctcttaata acttcgtata gcatacatta tacgaagtta tttcgaactc
      781 gagcggaatt ctagagtcgc ggccttaatt aattcgaagg ttatgcatgg tggtgatggt
      841 ggtgcgatcc cttcaggtcc tcctccgaga tcagcttttg ctcgtacgtc cgcggcttgt
      901 cgttttccag ggtctgccgg atcagctggg tgatctgcag ggtgtcgtag tacttctggg
      961 cctcattcac ggtgtagacc tcgatggctt cctgcttgcc cttgacctgg tgggctccca
     1021 ggttgatgaa ggtccactgg gcctggcagc acttcttcac gccggctgta aaggccaggg
     1081 cgtagggcag ctgtcttgtc agggcttcca gcctggcggc cacgttcacg gcgtccccca
     1141 gcagggtgtg gtccatcttc aggctgctgc ccatgttgcc ctcgatcacg tggccgtagc
     1201 tcaggccgat gccggtgtac agcaggtgca gggggttggt ggcctccacg tggtgccgca
     1261 gctgcttcag ctcggagatg atgtccaggc tggtccggat ggcggcgtcg ccctgctctt
     1321 tggtgaagct ggccatcacg cagtcgccga tgaacttggt cacttcgccg ccgtaggcgc
     1381 tgatgatccg ggtgcagatg ctgaagtacc ggttgaccag gatgaccacc tcgttcacgg
     1441 gcagcttctc ggtcagggtg ctgaaggcca ggatgtcgct gaagaagatg atcttctcga
     1501 ccagctgggg ctccacggtc agggggttga tgccctggtt gatcaggtag atcactctgg
     1561 cgggcatgta cttttccagc acccggtggc tctgggtgat ggtctgcagc aggctcttga
     1621 tgggctggat catcagctcg ctgttctcgt tcaggttgat ggttttcatg gcccagttgg
     1681 ggaacatccg gtcggtgatg tcgtactcgg ttttcaggca caggatgttg gtgtgccggt
     1741 cgtccaccag gattttcttg tacagcttgt ccaccttctc gttctcgcct tccaggatct
     1801 ggaagaacag gccctgcagg tacagcagca cgccggtgat gttgtcccgc tggttgttct
     1861 tgatggacac cttgccgatc cgctggatct cttccaggct caggtggccg ctgatcttgc
     1921 tgatgtacac cagccgcttc atcatggatc ccttgtacag ctcgtccatg ccgccggtgg
     1981 agtggcggcc ctcggagcgt tcgtactgtt ccaccacggt gtagtcctcg ttgtgggagg
     2041 tgatgtccaa cttgcggtcg acgttgtagg cgccgggcat ctgcacgggc ttcttggcct
     2101 tgtaggtggt cttgaagtcc gccaggtagc ggccgccgtc cttcaggcgc agggccatct
     2161 taatgtcgcc cttcagcacg ccgtcctcgg ggtacaaccg ctcggtggac gcttcccagc
     2221 ccattgtctt cttctgcatt acggggccgt caggagggaa gttggtgccg cggagcttca
     2281 ccttgtagat cagggtgccg tcctccaggg aggtgtcctg ggtcacggtc acggcgccgc
     2341 cgtcctcgaa gttcatcacg cgctcccact tgaagccctc ggggaaggac tgcttatagt
     2401 agtcggggat gtcggcgggg tgcttggtga aggccctgga gccgtacatg aactgagggg
     2461 acaggatgtc ccaggagaag ggcagggggc cacccttggt caccttcagc ttggcggtct
     2521 gggtgccctc gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg tggccgttca
     2581 tggagccctc catgtgcacc ttgaaccgca tgaactcctt gatcactgcc tcgcccttgc
     2641 tcaccatggt ggagccagtt aacccttcca tctgattgga gaaggaggtg ggcgcaccct
     2701 gttggccgta gccttgctgc ccatagtcgc cctgaggccc gtagccaccg ccaccgccgc
     2761 tggctggctg cccgtaatcg ggttgataac caccctgagg cccgtaggag tcctggggcc
     2821 catagcctcc ggggccctgc ccatagcccg catcgccgta ggcatcgcca ggagctggtt
     2881 gcttttctgg ggcgcctgga ggtgcgcgca tgaatggggc ggcccagcct gtctccttga
     2941 acacgaacca taggttgcca acccagagca ccaggttcag gaagccaaac accaccgagg
     3001 tgttgagtcc tgaagtcaca gggtccctca gttccttgca tgtgtttcct gtctggcggc
     3061 acataggcat ctccttgata atgttctctg ggtcagtggc catcttcaca tcggacaggc
     3121 ctttggccca ggcggatgag ctaactagcc acatgaaagc gaacactgct gtggccagga
     3181 agtccatcat tggccctttg ttgttctctc ggtacttgtt ctgcaggaag atgtaggtgg
     3241 ccagggcccc catggagtag aggaaggcaa acacagccac ggtgacaaag aattcagccg
     3301 aggaggagta gtcaccaact aggaagatct tggtagtgcc ccctttaacg caggagggtg
     3361 catcaaagta cacttggtgc agcctgaatg ggtactcaaa ttcgacttcg atgttgaggg
     3421 cactctccgt cttgttggca cactccacgc tcagccgaag ctctccggtg tagctgccgc
     3481 acgtagcaaa ggcgaagatg gcaaagaccc actgcagcac cttcacgaag ccaaggggct
     3541 ccttgaccac ccggaactga cccccagcca ccagctgatt caccacgtcc atgtctgcca
     3601 gcagcagcat ggtggcgcta gcatgtggat ccataacttc gtataaagta tcctatacga
     3661 agttatttgc cttaacccag aaattatcac tgttattctt tagaatggtg caaagaataa
     3721 cttcgtataa tgtatgctat acgaagttat gtcgagataa tcaacctctg gattacaaaa
     3781 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     3841 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     3901 tgtataaatc ctggttagtt cttgccacgg cggaactcat cgccgcctgc cttgcccgct
     3961 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgtttatt tgtgaaattt
     4021 gtgatgctat tgctttattt gtaaccattc tagctttatt tgtgaaattt gtgatgctat
     4081 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     4141 ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcacgt gcggaccgag
     4201 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     4261 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     4321 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     4381 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     4441 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4501 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4561 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4621 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4681 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4741 gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     4801 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4861 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     4921 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     4981 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     5041 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     5101 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     5161 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5221 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     5281 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5341 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5401 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5461 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5521 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5581 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5641 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5701 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5761 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5821 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5881 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5941 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6001 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6061 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6121 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     6181 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6241 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6301 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6361 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6421 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     6481 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6541 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6601 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6661 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6721 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6781 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6841 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6901 cctttttacg gttcctggcc ttttgc
//
