LOCUS       sequence-309694-        6915 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-conditional expression of synaptoPAC, a fusion of synaptophysin,
            mScarlet, and bPAC in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6915)
  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 6915)
  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..6915
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   20..149
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        181..628
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(643..676)
                     /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(727..760)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(816..833)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(843..872)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(1941..2636)
                     /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(2216..2235)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2485..2503
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    3622..3655
                     /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    3706..3739
                     /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(3801..3821)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4197..4337
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4197..4326
                     /label=AAV2 ITR
     rep_origin      4412..4867
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4499..4518)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4709..4730
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4884..4903)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5003..5025
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5063..5081)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5149..5253
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5254..5322,5323..6114)
                     /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(5472..5491)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6285..6873
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6774..6793
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
       61 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      121 agtggccaac tccatcacta ggggttcctg cggccgcacg cgtagagggc cctgcgtatg
      181 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
      241 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      301 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      361 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      421 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      481 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      541 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      601 aggagtcgtg tcgtgcctga gagcgcagtc gctagtaccg gtataacttc gtataggata
      661 ctttatacga agttatgcag aatggtagct ggattgtagc tgctattagc aatatgaaac
      721 ctcttaataa cttcgtatag catacattat acgaagttat ttcgaactcg agcggaattc
      781 tagagtcgcg gccttaatta attcgaaggt tatgcatggt ggtgatggtg gtgcgatccc
      841 ttcaggtcct cctccgagat cagcttttgc tcgtacgtcc gcggcttgtc gttttccagg
      901 gtctgccgga tcagctgggt gatctgcagg gtgtcgtagt acttctgggc ctcattcacg
      961 gtgtagacct cgatggcttc ctgcttgccc ttgacctggt gggctcccag gttgatgaag
     1021 gtccactggg cctggcagca cttcttcacg ccggctgtaa aggccagggc gtagggcagc
     1081 tgtcttgtca gggcttccag cctggcggcc acgttcacgg cgtcccccag cagggtgtgg
     1141 tccatcttca ggctgctgcc catgttgccc tcgatcacgt ggccgtagct caggccgatg
     1201 ccggtgtaca gcaggtgcag ggggttggtg gcctccacgt ggtgccgcag ctgcttcagc
     1261 tcggagatga tgtccaggct ggtccggatg gcggcgtcgc cctgctcttt ggtgaagctg
     1321 gccatcacgc agtcgccgat gaacttggtc acttcgccgc cgtaggcgct gatgatccgg
     1381 gtgcagatgc tgaagtaccg gttgaccagg atgaccacct cgttcacggg cagcttctcg
     1441 gtcagggtgc tgaaggccag gatgtcgctg aagaagatga tcttctcgac cagctggggc
     1501 tccacggtca gggggttgat gccctggttg atcaggtaga tcactctggc gggcatgtac
     1561 ttttccagca cccggtggct ctgggtgatg gtctgcagca ggctcttgat gggctggatc
     1621 atcagctcgc tgttctcgtt caggttgatg gttttcatgg cccagttggg gaacatccgg
     1681 tcggtgatgt cgtactcggt tttcaggcac aggatgttgg tgtgccggtc gtccaccagg
     1741 attttcttgt acagcttgtc caccttctcg ttctcgcctt ccaggatctg gaagaacagg
     1801 ccctgcaggt acagcagcac gccggtgatg ttgtcccgct ggttgttctt gatggacacc
     1861 ttgccgatcc gctggatctc ttccaggctc aggtggccgc tgatcttgct gatgtacacc
     1921 agccgcttca tcatggatcc cttgtacagc tcgtccatgc cgccggtgga gtggcggccc
     1981 tcggagcgtt cgtactgttc caccacggtg tagtcctcgt tgtgggaggt gatgtccaac
     2041 ttgcggtcga cgttgtaggc gccgggcatc tgcacgggct tcttggcctt gtaggtggtc
     2101 ttgaagtccg ccaggtagcg gccgccgtcc ttcaggcgca gggccatctt aatgtcgccc
     2161 ttcagcacgc cgtcctcggg gtacaaccgc tcggtggacg cttcccagcc cattgtcttc
     2221 ttctgcatta cggggccgtc aggagggaag ttggtgccgc ggagcttcac cttgtagatc
     2281 agggtgccgt cctccaggga ggtgtcctgg gtcacggtca cggcgccgcc gtcctcgaag
     2341 ttcatcacgc gctcccactt gaagccctcg gggaaggact gcttatagta gtcggggatg
     2401 tcggcggggt gcttggtgaa ggccctggag ccatacatga actgagggga caggatgtcc
     2461 caggagaagg gcagggggcc acccttggtc accttcagct tggcggtctg ggtgccctcg
     2521 taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcat ggagccctcc
     2581 atgtgcacct tgaaccgcat gaactccttg atcactgcct cgcccttgct caccatggtg
     2641 gagccagtta acccttccat ctgattggag aaggaggtgg gcgcaccctg ttggccgtag
     2701 ccttgctgcc catagtcgcc ctgaggcccg tagccaccgc caccgccgct ggctggctgc
     2761 ccgtaatcgg gttgataacc accctgaggc ccgtaggagt cctggggccc atagcctccg
     2821 gggccctgcc catagcccgc atcgccgtag gcatcgccag gagctggttg cttttctggg
     2881 gcgcctggag gtgcgcgcat gaatggggcg gcccagcctg tctccttgaa cacgaaccat
     2941 aggttgccaa cccagagcac caggttcagg aagccaaaca ccaccgaggt gttgagtcct
     3001 gaagtcacag ggtccctcag ttccttgcat gtgtttcctg tctggcggca cataggcatc
     3061 tccttgataa tgttctctgg gtcagtggcc atcttcacat cggacaggcc tttggcccag
     3121 gcggatgagc taactagcca catgaaagcg aacactgctg tggccaggaa gtccatcatt
     3181 ggccctttgt tgttctctcg gtacttgttc tgcaggaaga tgtaggtggc cagggccccc
     3241 atggagtaga ggaaggcaaa cacagccacg gtgacaaaga attcagccga ggaggagtag
     3301 tcaccaacta ggaagatctt ggtagtgccc cctttaacgc aggagggtgc atcaaagtac
     3361 acttggtgca gcctgaatgg gtactcaaat tcgacttcga tgttgagggc actctccgtc
     3421 ttgttggcac actccacgct cagccgaagc tctccggtgt agctgccgca cgtagcaaag
     3481 gcgaagatgg caaagaccca ctgcagcacc ttcacgaagc caaggggctc cttgaccacc
     3541 cggaactgac ccccagccac cagctgattc accacgtcca tgtctgccag cagcagcatg
     3601 gtggcgctag catgtggatc cataacttcg tataaagtat cctatacgaa gttatttgcc
     3661 ttaacccaga aattatcact gttattcttt agaatggtgc aaagaataac ttcgtataat
     3721 gtatgctata cgaagttatg tcgagataat caacctctgg attacaaaat ttgtgaaaga
     3781 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     3841 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     3901 tggttagttc ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     3961 gctcggctgt tgggcactga caattccgtg gtgtttattt gtgaaatttg tgatgctatt
     4021 gctttatttg taaccatcta gctttatttg tgaaatttgt gatgctattg ctttatttgt
     4081 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     4141 ggttcagggg gagatgtggg aggtttttta aagcacggtg cggaccgagc ggccgcagga
     4201 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     4261 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     4321 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4381 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     4441 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     4501 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4561 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4621 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4681 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4741 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     4801 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     4861 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     4921 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4981 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     5041 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     5101 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5161 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5221 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5281 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5341 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5401 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5461 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5521 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5581 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5641 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5701 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5761 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5821 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5881 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5941 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6001 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6061 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6121 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6181 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6241 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6301 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6361 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6421 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6481 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6541 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6601 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6661 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6721 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6781 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6841 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6901 ttcctggcct tttgc
//
