LOCUS       sequence-378208-        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     38..59
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(213..232)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     332..354
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(392..410)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        478..582
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(583..651,652..1443)
                     /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      1614..2202
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2103..2122
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2264..2393
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        2438..2885
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    2923..2956
                     /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(5284..5979)
                     /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(5559..5578)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5828..5846
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      5976..5985
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(5992..6025)
                     /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    6143..6619
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(6342..6361)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   6659..6799
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6659..6788
                     /label=AAV2 ITR
     rep_origin      join(6874..7133,1..196)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6961..6980)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
       61 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
      121 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
      181 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
      241 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
      301 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
      361 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
      421 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
      481 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
      541 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
      601 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
      661 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
      721 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
      781 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
      841 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
      901 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
      961 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     1021 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     1081 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     1141 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     1201 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     1261 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     1321 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     1381 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     1441 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     1501 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     1561 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     1621 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     1681 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     1741 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     1801 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     1861 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     1921 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     1981 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2041 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2101 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2161 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2221 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc
     2281 gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
     2341 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg
     2401 cacgcgtgtg tctagactgc agagggccct gcgtatgagt gcaagtgggt tttaggacca
     2461 ggatgaggcg gggtgggggt gcctacctga cgaccgaccc cgacccactg gacaagcacc
     2521 caacccccat tccccaaatt gcgcatcccc tatcagagag ggggagggga aacaggatgc
     2581 ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac agtgccttcg cccccgcctg
     2641 gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc gccggtcccc
     2701 cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc cgccgccggc ccagccggac
     2761 cgcaccacgc gaggcgcgag ataggggggc acgggcgcga ccatctgcgc tgcggcgccg
     2821 gcgactcagc gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag
     2881 cgcagtcgag aaggtaccgg atcctctaga gtcgactccg gagaagttcc tattctctag
     2941 aaagtatagg aacttcgcag aatggtagct ggattgtagc tgctattagc aatatgaaac
     3001 ctcttagaag ttcctattct tcaaaaggta taggaacttc ggcgcgcctc atagccgtcc
     3061 gatgaccatg acgtccacca cctcgccttt gtgaagttcc acatactgct ctgtctttgg
     3121 aggtagcatc agcaatccat tggcactgcg catgctcatc agacggctgc tcatctgatt
     3181 acctgtactc tgggcccagg gcagtggttc ttggtgatgc caagtcagta tacaccggtg
     3241 gtattctggg cgagggtcca gctttacgtc acatgataac ctagctttga tgatggttgg
     3301 ccgaggatcc aagataccct gcatttttct cagtgcgggt acaacaaaga ggttgcaggt
     3361 gaccacagct gatacaggat tccctggtag tgcaaaaata atttttctta caccatcaat
     3421 atccaaagtt gcaaatgttg ttggcaagcc tggcttcata aaaaccctgc caaaatggat
     3481 ctgagcatga agatcgatgt ccagcacctg cttgagatag tccttttccc ccatggatac
     3541 acctcctgac gtgatgatga catcagcacg gctgataccc tcattcaagg cattgagtaa
     3601 gtcatctggg ttgtctccta caattcccaa gttgattgtt gggtaaccat gttcctgaat
     3661 tgttgctaga agtgttgatc gattgctgtc ccgaatcttt cctggtaaga ggtcatcttc
     3721 aggatttagt agctcattcc ctgttgacat aacggcaacc actggaaact tattaacttc
     3781 aacttctgtg acacctacag ttgcaaggag accaatttct gaagggccca tatgggttcc
     3841 ttttgccaaa acacactccc ctcttttaat gtcgtggccg atgggtctga tgtcttggcc
     3901 tggccgagct tgcaccagaa ttcgcacttc aagttcttca gtaccatcat ctgattccct
     3961 gatgagttca gtatcttcaa cttgcactac tgcatcagca ccgcagggga ttggagcacc
     4021 tgttgtaacc cgcatcactt gtccgggcat tactgtctga gttggctgtt caccagcttg
     4081 ggattcccca atgatgaaac gatctcctgg accatcagca gctcgaacag catagccatc
     4141 ttttactgat gctgggaatg ggggtaggtt gtcttttgca tacacatctt gagcaaggac
     4201 tcgccccatt ccatctcggt aattgatgat ttctgtacca agcaccggag tcatctccag
     4261 aactgtaatg aaggctttgt ccatagacgt cagaggaaaa ggagacatgc gatgtcttct
     4321 agccaccttg gtgatatcta cagcactgtg actggctctt agaatgttct ccttgctgct
     4381 gcacctggac tggacttttg gtgttggaca agaagctgta gagaggcgag atgtagcctg
     4441 ggcacggggc gactctgaag gagtagtgct aagggaggct gtgtctcgtg ggagcacctg
     4501 aacgccacga gaaatgatgg agtctggaat ctttgcagca agagctgctg cagtaatatg
     4561 tgatgaggaa ctatcttctg tcgaagctac accactgtct ttcttttctt cttcctcctc
     4621 ttcacactga actcctttgt cttctgtctg cttatgtggg ctagttgtag gaggtggaga
     4681 aagaggaggt ggtggggaag gtaaatcttc aagttcatca tgcacctcct ttacttttac
     4741 aatggcatca cgtaaaaggt caatggcatg aggtagagct ggcagtatga actgaaagca
     4801 ttcctgagat cctttcttgc taccaggcag gttaattatc agagttttcc ctcttattcc
     4861 acacactggt ctagagagca tgcccagagg tgtaacatta agtgatccca tcagcattgc
     4921 cagggccatc cctggcgctt cccgttctat tacttctttt gtggcctctg gagtgacatc
     4981 tcgtggtgca aaccctgttc cgccagttgt taatatcaaa ttaagttcct tctcatcaca
     5041 ccagtctatg agtgtttcct tgatttcttc tatttcatct ggtactatct tgtatgctga
     5101 tatagtccca cccaacaaag aaggatcctg gacgagatct tttagattta tcccactgcg
     5161 gtcttctgca agattcctga agcagctatc actcactgtg aggactccga cacggatttg
     5221 atggtcgtgg tttgtgagga tcattccctc ggtcgccatg ctcgagatct gagtccggcc
     5281 ggacttgtac agctcgtcca tgccgccggt ggagtggcgg ccctcggagc gttcgtactg
     5341 ttccaccacg gtgtagtcct cgttgtggga ggtgatgtcc aacttgcggt ccacgttgta
     5401 ggcgccgggc atctgcacgg gcttcttggc cttgtaggtg gtcttgaagt ccgccaggta
     5461 gcgcccgccg tccttcaggc gcagggccat cttaatgtcg cccttcagca cgccgtcctc
     5521 ggggtacaac cgctcggtgg acgcttccca gcccattgtc ttcttctgca ttacggggcc
     5581 gtcaggaggg aagttggtgc cgcggagctt caccttgtag atcagggtgc cgtcctccag
     5641 ggaggtgtcc tgggtcacgg tcacggcgcc gccgtcctcg aagttcatca cgcgctccca
     5701 cttgaagccc tcggggaagg actgcttata gtagtcgggg atgtcggcgg ggtgcttggt
     5761 gaaggccctg gagccgtaca tgaactgagg ggacaggatg tcccaggaga agggcagggg
     5821 gccacccttg gtcaccttca gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc
     5881 ctcgccctcg atctcgaact cgtggccgtt catggagccc tccatgtgca ccttgaaccg
     5941 catgaactcc ttgatcactg cctcgccctt gctcaccatg gtggcgctag cgaagttcct
     6001 atactttcta gagaatagga acttcttgcc ttaacccaga aattatcact gttattcttt
     6061 agaatggtgc aaagagaagt tcctatacct tttgaagaat aggaacttcg aattcgatat
     6121 caagcttatc gatgagatct acgggtggca tccctgtgac ccctccccag tgcctctcct
     6181 ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat
     6241 cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag
     6301 caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg
     6361 gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct
     6421 cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt
     6481 tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta
     6541 atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac
     6601 tgctcccttc cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag
     6661 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     6721 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     6781 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     6841 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     6901 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
     6961 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     7021 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     7081 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtt
//
