LOCUS       sequence-310531-        6645 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 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 6645)
  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 6645)
  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-synaptoPAC-minWPRE
FEATURES             Location/Qualifiers
     source          1..6645
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   71..211
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   71..200
                     /label=AAV2 ITR
     rep_origin      286..741
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(373..392)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     583..604
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(758..777)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     877..899
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(937..955)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1023..1127
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1128..1196,1197..1988)
                     /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(1346..1365)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2159..2747
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2648..2667
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2809..2938
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        2970..3417
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             4395..5090
                     /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(4528..4546)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4796..4815
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             6159..6188
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             6198..6215
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6320..6340)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcac ggtgcggacc
       61 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
      121 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
      181 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
      241 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
      301 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
      361 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
      421 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
      481 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
      541 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
      601 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
      661 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
      721 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
      781 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
      841 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
      901 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
      961 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     1021 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1081 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     1141 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     1201 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     1261 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     1321 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     1381 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     1441 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     1501 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     1561 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     1621 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     1681 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     1741 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     1801 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     1861 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     1921 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     1981 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     2041 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     2101 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2161 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2221 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2281 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     2341 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2401 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2461 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2521 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2581 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2641 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2701 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2761 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc tgcaggcagc
     2821 tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc
     2881 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc
     2941 ggccgcacgc gtagagggcc ctgcgtatga gtgcaagtgg gttttaggac caggatgagg
     3001 cggggtgggg gtgcctacct gacgaccgac cccgacccac tggacaagca cccaaccccc
     3061 attccccaaa ttgcgcatcc cctatcagag agggggaggg gaaacaggat gcggcgaggc
     3121 gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg
     3181 cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact
     3241 ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg accgcaccac
     3301 gcgaggcgcg agataggggg gcacgggcgc gaccatctgc gctgcggcgc cggcgactca
     3361 gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg
     3421 ctagcgccac catgctgctg ctggcagaca tggacgtggt gaatcagctg gtggctgggg
     3481 gtcagttccg ggtggtcaag gagccccttg gcttcgtgaa ggtgctgcag tgggtctttg
     3541 ccatcttcgc ctttgctacg tgcggcagct acaccggaga gcttcggctg agcgtggagt
     3601 gtgccaacaa gacggagagt gccctcaaca tcgaagtcga atttgagtac ccattcaggc
     3661 tgcaccaagt gtactttgat gcaccctcct gcgttaaagg gggcactacc aagatcttcc
     3721 tagttggtga ctactcctcc tcggctgaat tctttgtcac cgtggctgtg tttgccttcc
     3781 tctactccat gggggccctg gccacctaca tcttcctgca gaacaagtac cgagagaaca
     3841 acaaagggcc aatgatggac ttcctggcca cagcagtgtt cgctttcatg tggctagtta
     3901 gctcatccgc ctgggccaaa ggcctgtccg atgtgaagat ggccactgac ccagagaaca
     3961 ttatcaagga gatgcctatg tgccgccaga caggaaacac atgcaaggaa ctgagggacc
     4021 ctgtgacttc aggactcaac acctcggtgg tgtttggctt cctgaacctg gtgctctggg
     4081 ttggcaacct atggttcgtg ttcaaggaga caggctgggc cgccccattc atgcgcgcac
     4141 ctccaggcgc cccagaaaag caaccagctc ctggcgatgc ctacggcgat gcgggctatg
     4201 ggcagggccc cggaggctat gggccccagg actcctacgg gcctcagggt ggttatcaac
     4261 ccgattacgg gcagccagcc agcggcggtg gcggtggcta cgggcctcag ggcgactatg
     4321 ggcagcaagg ctacggccaa cagggtgcgc ccacctcctt ctccaatcag atggaagggt
     4381 taactggctc caccatggtg agcaagggcg aggcagtgat caaggagttc atgcggttca
     4441 aggtgcacat ggagggctcc atgaacggcc acgagttcga gatcgagggc gagggcgagg
     4501 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     4561 ccttctcctg ggacatcctg tcccctcagt tcatgtatgg ctccagggcc ttcaccaagc
     4621 accccgccga catccccgac tactataagc agtccttccc cgagggcttc aagtgggagc
     4681 gcgtgatgaa cttcgaggac ggcggcgccg tgaccgtgac ccaggacacc tccctggagg
     4741 acggcaccct gatctacaag gtgaagctcc gcggcaccaa cttccctcct gacggccccg
     4801 taatgcagaa gaagacaatg ggctgggaag cgtccaccga gcggttgtac cccgaggacg
     4861 gcgtgctgaa gggcgacatt aagatggccc tgcgcctgaa ggacggcggc cgctacctgg
     4921 cggacttcaa gaccacctac aaggccaaga agcccgtgca gatgcccggc gcctacaacg
     4981 tcgaccgcaa gttggacatc acctcccaca acgaggacta caccgtggtg gaacagtacg
     5041 aacgctccga gggccgccac tccaccggcg gcatggacga gctgtacaag ggatccatga
     5101 tgaagcggct ggtgtacatc agcaagatca gcggccacct gagcctggaa gagatccagc
     5161 ggatcggcaa ggtgtccatc aagaacaacc agcgggacaa catcaccggc gtgctgctgt
     5221 acctgcaggg cctgttcttc cagatcctgg aaggcgagaa cgagaaggtg gacaagctgt
     5281 acaagaaaat cctggtggac gaccggcaca ccaacatcct gtgcctgaaa accgagtacg
     5341 acatcaccga ccggatgttc cccaactggg ccatgaaaac catcaacctg aacgagaaca
     5401 gcgagctgat gatccagccc atcaagagcc tgctgcagac catcacccag agccaccggg
     5461 tgctggaaaa gtacatgccc gccagagtga tctacctgat caaccagggc atcaaccccc
     5521 tgaccgtgga gccccagctg gtcgagaaga tcatcttctt cagcgacatc ctggccttca
     5581 gcaccctgac cgagaagctg cccgtgaacg aggtggtcat cctggtcaac cggtacttca
     5641 gcatctgcac ccggatcatc agcgcctacg gcggcgaagt gaccaagttc atcggcgact
     5701 gcgtgatggc cagcttcacc aaagagcagg gcgacgccgc catccggacc agcctggaca
     5761 tcatctccga gctgaagcag ctgcggcacc acgtggaggc caccaacccc ctgcacctgc
     5821 tgtacaccgg catcggcctg agctacggcc acgtgatcga gggcaacatg ggcagcagcc
     5881 tgaagatgga ccacaccctg ctgggggacg ccgtgaacgt ggccgccagg ctggaagccc
     5941 tgacaagaca gctgccctac gccctggcct ttacagccgg cgtgaagaag tgctgccagg
     6001 cccagtggac cttcatcaac ctgggagccc accaggtcaa gggcaagcag gaagccatcg
     6061 aggtctacac cgtgaatgag gcccagaagt actacgacac cctgcagatc acccagctga
     6121 tccggcagac cctggaaaac gacaagccgc ggacgtacga gcaaaagctg atctcggagg
     6181 aggacctgaa gggatcgcac caccatcacc accatgcata accttcgaat taattaaggc
     6241 cgcgactcta gaattccgct cgagataatc aacctctgga ttacaaaatt tgtgaaagat
     6301 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     6361 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     6421 ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     6481 ctcggctgtt gggcactgac aattccgtgg tgtttatttg tgaaatttgt gatgctattg
     6541 ctttatttgt aaccatctag ctttatttgt gaaatttgtg atgctattgc tttatttgta
     6601 accattataa gctgcaataa acaagttaac aacaacaatt gcatt
//
