LOCUS       sequence-403385-        6332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Flp-dependent enzymatically inactive Cre and mScarlet-KASH 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6332)
  AUTHORS   Roethler O, Zohar E, Cohen-Kashi Malina K, Bitan L, Gabel HW, 
            Spiegel I
  TITLE     Single genomic enhancers drive experience-dependent GABAergic 
            plasticity to maintain sensory processing in the adult cortex.
  JOURNAL   Neuron. 2023 Jun 20:S0896-6273(23)00399-9. doi: 
            10.1016/j.neuron.2023.05.026.
  PUBMED    37354902
REFERENCE   2  (bases 1 to 6332)
  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"; journalName: "Neuron. 
            2023 Jun 20:S0896-6273(23)00399-9. doi: 
            10.1016/j.neuron.2023.05.026."
COMMENT     Sequence Label: pAAV-Syn1-fDIO-mScarlet-KASH-2a-dCre
FEATURES             Location/Qualifiers
     source          1..6332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        9..456
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    487..520
                     /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(979..999)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(1015..1068)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(1342..2037)
                     /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(1617..1636)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1886..1904
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(2038..2067)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      2067..2076
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2083..2116)
                     /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)"
     misc_feature    2225..2813
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2278..2298)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2696..2707)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2845..3321
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3044..3063)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3361..3501
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3361..3490
                     /label=AAV2 ITR
     rep_origin      3576..4031
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3663..3682)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3873..3894
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4048..4067)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4167..4189
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4227..4245)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4313..4417
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4418..4486,4487..5278)
                     /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(4636..4655)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5449..6037
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5938..5957
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6099..6228
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc ggggtggggg tgcctacctg
       61 acgaccgacc ccgacccact ggacaagcac ccaaccccca ttccccaaat tgcgcatccc
      121 ctatcagaga gggggagggg aaacaggatg cggcgaggcg cgtgcgcact gccagcttca
      181 gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg cctcagcact
      241 gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc cccttcccgg ccaccttggt
      301 cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga gatagggggg
      361 cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg
      421 ggcagcggag gagtcgtgtc gtgcctgaga gcgcagggta ccggatcctc tagagtcgac
      481 tccggagaag ttcctattct ctagaaagta taggaacttc gcagaatggt agctggattg
      541 tagctgctat tagcaatatg aaacctctta gaagttccta ttcttcaaaa ggtataggaa
      601 cttcggcgcg ccctacttac ggattcgccg cataaccagt gaaacagcat tgctgtcact
      661 tggtcgtggc agcccggacc gacgatgaag catgtttagc tggcccaaat gttgctggat
      721 agtttttact gccagaccgc gcgcctgaag atatagaaga taatcgcgaa catcttcagg
      781 ttctgcggga aaccatttcc ggttattcaa cttgcaccat gccgcccacg accggcaaac
      841 ggacagaagc attttccagg tatgctcaga aaacgcctgg cgatccctga acatgtccat
      901 caggttcttg cgaacctcat cactcgttgc atcgaccggt aatgcaggca aattttggtg
      961 tacggtcagt aaattggaca ccttcctctt cttcttgggg gttctgaaag cgcttgggcc
     1021 aggattctcc tcgacgtcac cgcatgttag cagacttcct ctgccctctc cactgccggt
     1081 gggaggtggc ccgttggtgt accgcagcat ggggtagaag gatcgggcaa agttgttggc
     1141 ctgggtgcag ctgtagtcat cttcagaggc aggtagcagg caggccagga gcagcagcag
     1201 cagcagaagc agctgcaacg gtagcgctgc cctgatcacc cttgagagga aggagcgtct
     1261 cggctgggag ctgccggggc tgtcgaggtg gggcatcctg ctgtctgtct cctcctcctc
     1321 ctctcgagat ctgagtccgg acttgtacag ctcgtccatg ccgccggtgg agtggcggcc
     1381 ctcggagcgt tcgtactgtt ccaccacggt gtagtcctcg ttgtgggagg tgatgtccaa
     1441 cttgcggtcg acgttgtagg cgccgggcat ctgcacgggc ttcttggcct tgtaggtggt
     1501 cttgaagtcc gccaggtagc gtccgccgtc cttcaggcgc agggccatct taatgtcgcc
     1561 cttcagcacg ccgtcctcgg ggtacaaccg ctcggtggac gcttcccagc ccattgtctt
     1621 cttctgcatt acggggccgt caggagggaa gttggtgccg cggagcttca ccttgtagat
     1681 cagggtgccg tcctccaggg aggtgtcctg ggtcacggtc acggcgccgc cgtcctcgaa
     1741 gttcatcacg cgctcccact tgaagccctc ggggaaggac tgcttatagt agtcggggat
     1801 gtcggcgggg tgcttggtga aggccctgga gccgtacatg aactgagggg acaggatgtc
     1861 ccaggagaag ggcagggggc cacccttggt caccttcagc ttggcggtct gggtgccctc
     1921 gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg tggccgttca tggagccctc
     1981 catgtgcacc ttgaaccgca tgaactcctt gatcactgcc tcgcccttgc tcaccatcag
     2041 atcttcttcg ctaatcagtt tctgttccat ggtggcgcta gcgaagttcc tatactttct
     2101 agagaatagg aacttcttgc cttaacccag aaattatcac tgttattctt tagaatggtg
     2161 caaagagaag ttcctatacc ttttgaagaa taggaacttc gaattcgata tcaagcttat
     2221 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     2281 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     2341 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     2401 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     2461 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     2521 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     2581 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     2641 gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     2701 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     2761 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     2821 ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc
     2881 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     2941 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     3001 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     3061 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     3121 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     3181 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     3241 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     3301 actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc
     3361 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     3421 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     3481 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     3541 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     3601 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     3661 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3721 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3781 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3841 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3901 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     3961 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4021 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4081 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4141 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4201 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4261 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4321 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4381 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4441 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4501 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4561 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4621 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4681 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4741 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4801 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4861 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4921 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4981 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5041 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5101 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5161 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5221 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5281 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5341 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5401 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5461 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5521 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5581 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     5641 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     5701 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     5761 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     5821 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     5881 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     5941 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6001 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6061 acggttcctg gccttttgct ggccttttgc tcacatgtcc tgcaggcagc tgcgcgctcg
     6121 ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg
     6181 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc ggccgcacgc
     6241 gtaagctttg caaagatgga taaagtttta aacagagagg aatctttgca gctaatggac
     6301 cttctaggtc ttgaaaggag tgggaattgg ct
//
