LOCUS       sequence-384098-        7133 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector to drive the Flp-dependent expression of mScarlet-Gphn 
            (isoform P1) under the control of human Synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7133)
  AUTHORS   Liebsch F, Eggersmann FR, Merkler Y, Kloppenburg P, Schwarz G
  TITLE     Automated Image Analysis Reveals Different Localization of Synaptic 
            Gephyrin C4 Splice Variants.
  JOURNAL   eNeuro. 2023 Jan 4;10(1):ENEURO.0102-22.2022. doi: 
            10.1523/ENEURO.0102-22.2022. Print 2023 Jan.
  PUBMED    36543537
REFERENCE   2  (bases 1 to 7133)
  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: 
            "10.1523/ENEURO.0102-22.2022"; journalName: "eNeuro"; date: 
            "2023-01-4- 4"; volume: "10"; issue: "1"; pages: 
            "ENEURO.0102-22.2022"
COMMENT     Sequence Label: pAAV-hSyn-fDIO-mScarlet-Gphn_P1
FEATURES             Location/Qualifiers
     source          1..7133
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     39..60
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(214..233)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     333..355
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(393..411)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        479..583
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(584..652,653..1444)
                     /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"
     rep_origin      1615..2203
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2104..2123
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2265..2394
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        2439..2886
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    2924..2957
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             complement(5285..5980)
                     /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(5560..5579)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5829..5847
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      5977..5986
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(5993..6026)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     polyA_signal    6144..6620
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(6343..6362)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   6660..6800
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6660..6789
                     /label=AAV2 ITR
     rep_origin      join(6875..7133,1..197)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6962..6981)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
       61 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
      121 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
      181 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
      241 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
      301 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
      361 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
      421 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
      481 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
      541 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
      601 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
      661 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
      721 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
      781 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
      841 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
      901 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
      961 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     1021 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     1081 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     1141 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     1201 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     1261 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     1321 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     1381 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     1441 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     1501 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     1561 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     1621 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     1681 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     1741 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     1801 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     1861 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     1921 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     1981 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     2041 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     2101 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     2161 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     2221 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtcctgca ggcagctgcg
     2281 cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag
     2341 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc
     2401 gcacgcgtgt gtctagactg cagagggccc tgcgtatgag tgcaagtggg ttttaggacc
     2461 aggatgaggc ggggtggggg tgcctacctg acgaccgacc ccgacccact ggacaagcac
     2521 ccaaccccca ttccccaaat tgcgcatccc ctatcagaga gggggagggg aaacaggatg
     2581 cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga cagtgccttc gcccccgcct
     2641 ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc tgacgtcact cgccggtccc
     2701 ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg cccagccgga
     2761 ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg accatctgcg ctgcggcgcc
     2821 ggcgactcag cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc gtgcctgaga
     2881 gcgcagtcga gaaggtaccg gatcctctag agtcgactcc ggagaagttc ctattctcta
     2941 gaaagtatag gaacttcgca gaatggtagc tggattgtag ctgctattag caatatgaaa
     3001 cctcttagaa gttcctattc ttcaaaaggt ataggaactt cggcgcgcct catagccgtc
     3061 cgatgaccat gacgtccacc acctcgcctt tgtgaagttc cacatactgc tctgtctttg
     3121 gaggtagcat cagcaatcca ttggcactgc gcatgctcat cagacggctg ctcatctgat
     3181 tacctgtact ctgggcccag ggcagtggtt cttggtgatg ccaagtcagt atacaccggt
     3241 ggtattctgg gcgagggtcc agctttacgt cacatgataa cctagctttg atgatggttg
     3301 gccgaggatc caagataccc tgcatttttc tcagtgcggg tacaacaaag aggttgcagg
     3361 tgaccacagc tgatacagga ttccctggta gtgcaaaaat aatttttctt acaccatcaa
     3421 tatccaaagt tgcaaatgtt gttggcaagc ctggcttcat aaaaaccctg ccaaaatgga
     3481 tctgagcatg aagatcgatg tccagcacct gcttgagata gtccttttcc cccatggata
     3541 cacctcctga cgtgatgatg acatcagcac ggctgatacc ctcattcaag gcattgagta
     3601 agtcatctgg gttgtctcct acaattccca agttgattgt tgggtaacca tgttcctgaa
     3661 ttgttgctag aagtgttgat cgattgctgt cccgaatctt tcctggtaag aggtcatctt
     3721 caggatttag tagctcattc cctgttgaca taacggcaac cactggaaac ttattaactt
     3781 caacttctgt gacacctaca gttgcaagga gaccaatttc tgaagggccc atatgggttc
     3841 cttttgccaa aacacactcc cctcttttaa tgtcgtggcc gatgggtctg atgtcttggc
     3901 ctggccgagc ttgcaccaga attcgcactt caagttcttc agtaccatca tctgattccc
     3961 tgatgagttc agtatcttca acttgcacta ctgcatcagc accgcagggg attggagcac
     4021 ctgttgtaac ccgcatcact tgtccgggca ttactgtctg agttggctgt tcaccagctt
     4081 gggattcccc aatgatgaaa cgatctcctg gaccatcagc agctcgaaca gcatagccat
     4141 cttttactga tgctgggaat gggggtaggt tgtcttttgc atacacatct tgagcaagga
     4201 ctcgccccat tccatctcgg taattgatga tttctgtacc aagcaccgga gtcatctcca
     4261 gaactgtaat gaaggctttg tccatagacg tcagaggaaa aggagacatg cgatgtcttc
     4321 tagccacctt ggtgatatct acagcactgt gactggctct tagaatgttc tccttgctgc
     4381 tgcacctgga ctggactttt ggtgttggac aagaagctgt agagaggcga gatgtagcct
     4441 gggcacgggg cgactctgaa ggagtagtgc taagggaggc tgtgtctcgt gggagcacct
     4501 gaacgccacg agaaatgatg gagtctggaa tctttgcagc aagagctgct gcagtaatat
     4561 gtgatgagga actatcttct gtcgaagcta caccactgtc tttcttttct tcttcctcct
     4621 cttcacactg aactcctttg tcttctgtct gcttatgtgg gctagttgta ggaggtggag
     4681 aaagaggagg tggtggggaa ggtaaatctt caagttcatc atgcacctcc tttactttta
     4741 caatggcatc acgtaaaagg tcaatggcat gaggtagagc tggcagtatg aactgaaagc
     4801 attcctgaga tcctttcttg ctaccaggca ggttaattat cagagttttc cctcttattc
     4861 cacacactgg tctagagagc atgcccagag gtgtaacatt aagtgatccc atcagcattg
     4921 ccagggccat ccctggcgct tcccgttcta ttacttcttt tgtggcctct ggagtgacat
     4981 ctcgtggtgc aaaccctgtt ccgccagttg ttaatatcaa attaagttcc ttctcatcac
     5041 accagtctat gagtgtttcc ttgatttctt ctatttcatc tggtactatc ttgtatgctg
     5101 atatagtccc acccaacaaa gaaggatcct ggacgagatc ttttagattt atcccactgc
     5161 ggtcttctgc aagattcctg aagcagctat cactcactgt gaggactccg acacggattt
     5221 gatggtcgtg gtttgtgagg atcattccct cggtcgccat gctcgagatc tgagtccggc
     5281 cggacttgta cagctcgtcc atgccgccgg tggagtggcg gccctcggag cgttcgtact
     5341 gttccaccac ggtgtagtcc tcgttgtggg aggtgatgtc caacttgcgg tccacgttgt
     5401 aggcgccggg catctgcacg ggcttcttgg ccttgtaggt ggtcttgaag tccgccaggt
     5461 agcgcccgcc gtccttcagg cgcagggcca tcttaatgtc gcccttcagc acgccgtcct
     5521 cggggtacaa ccgctcggtg gacgcttccc agcccattgt cttcttctgc attacggggc
     5581 cgtcaggagg gaagttggtg ccgcggagct tcaccttgta gatcagggtg ccgtcctcca
     5641 gggaggtgtc ctgggtcacg gtcacggcgc cgccgtcctc gaagttcatc acgcgctccc
     5701 acttgaagcc ctcggggaag gactgcttat agtagtcggg gatgtcggcg gggtgcttgg
     5761 tgaaggccct ggagccgtac atgaactgag gggacaggat gtcccaggag aagggcaggg
     5821 ggccaccctt ggtcaccttc agcttggcgg tctgggtgcc ctcgtagggg cggccctcgc
     5881 cctcgccctc gatctcgaac tcgtggccgt tcatggagcc ctccatgtgc accttgaacc
     5941 gcatgaactc cttgatcact gcctcgccct tgctcaccat ggtggcgcta gcgaagttcc
     6001 tatactttct agagaatagg aacttcttgc cttaacccag aaattatcac tgttattctt
     6061 tagaatggtg caaagagaag ttcctatacc ttttgaagaa taggaacttc gaattcgata
     6121 tcaagcttat cgatgagatc tacgggtggc atccctgtga cccctcccca gtgcctctcc
     6181 tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca
     6241 tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga
     6301 gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct
     6361 ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc
     6421 tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat
     6481 ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct
     6541 aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca
     6601 ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg agcggccgca
     6661 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     6721 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     6781 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     6841 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     6901 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     6961 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     7021 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     7081 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggt
//
