LOCUS       sequence-429605-        6404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetically encoded fluorescent probe for labelling endogenous 
            excitatory synapses on PV interneurons in vivo.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6404)
  AUTHORS   Okur Z, Schlauri N, Bitsikas V, Panopoulou M, Karmakar K, Schreiner 
            D, Scheiffele P
  TITLE     Control of neuronal excitation-inhibition balance by BMP-SMAD1 
            signaling
  JOURNAL   bioRxiv 2023
REFERENCE   2  (bases 1 to 6404)
  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: "bioRxiv 
            2023"
COMMENT     Sequence Label: pAAV-S5E2-FingR-PSD95-mGL
FEATURES             Location/Qualifiers
     source          1..6404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    798..809
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     intron          1594..1726
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     CDS             2014..2037
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(2068..2070,2071..2781)
                     /codon_start=1
                     /product="monomeric GFP derivative that displays 
                     exceptionally high brightness in cells (Campbell et al., 
                     2020)"
                     /label=mGreenLantern
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLGYGVACFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIVLKGIDFKEDGNILGHKLEYNFNSHKVYITADKQKNGIKA
                     NFKTRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLKE
                     RVTAAGITHDMDELYK"
     primer_bind     complement(2110..2131)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2833..3174
                     /codon_start=1
                     /product="zinc-finger domain from the CCR5-224 (+) 
                     zinc-finger nuclease that recognizes the left half-site 
                     target sequence in the human CCR5 gene (Perez et al., 2008;
                     Gross et al., 2013)"
                     /label=CCR5TC
                     /translation="FQCRICMRNFSDRSNLSRHIRTHTGEKPFACDICGRKFAISSNLN
                     SHTKIHTGSQKPFQCRICMRNFSRSDNLARHIRTHTGEKPFACDICGRKFATSGNLTRH
                     TKIHLRGSQL"
     CDS             3223..3348
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the rat zinc finger protein Kid-1 
                     (Witzgall et al., 1994)"
                     /label=KRAB-A
                     /translation="VSVTFEDVAVLFTRDEWKKLDLSQRSLYREVMLENYSNLASM"
     misc_feature    3400..3988
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3453..3473)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3871..3882)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4157..4238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4194..4213)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4248..4267
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4317..4457
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4317..4446
                     /label=AAV2 ITR
     rep_origin      4532..4987
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4619..4638)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4829..4850
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5004..5023)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5123..5145
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5183..5201)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5269..5373
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5374..5442,5443..6234)
                     /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(5592..5611)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gcgtagagtc atcctcatcg tcactagtgg tttaatctaa catggctgct
      841 atagcttact gactagaagt taagtgcaca cttcctaaaa gaaggctttg acacaagcca
      901 cttcagttcc ctcctcattt tcttgtcccc attcctctct ctgtagaatt ctgagatttc
      961 aattcagttt tatacagaaa ccacattact gtaagcccta caaagttatg gcaatatagc
     1021 tatatggagt caagtaatgt aggttatttt tttcccaatg gtgctggtga aggtggcaat
     1081 tatgtagcta tacttagcag actgaggaaa ttctgctaga gtcagcattt gtctcttcat
     1141 tgctatgaaa cagtaatgga aaaataaaca aaaacaaaag gcaaacacta tgcataattc
     1201 cctcagatca tattaacatg tgatgttgga gtaaattgtt ataaccccat tttggaaata
     1261 cttaccttaa ttaactatga tttccttaaa ataatgcagt atttacaatc tatatgaaag
     1321 cactatatgg gacacatggt atgatggaac agtgcaccca agagacacca agaacattcc
     1381 tgtctgtggc agtcttttct ctatacagag gcatttagtc tcaattgctc agagttattt
     1441 taaacttcct gcagcccggg ctgggcataa aagtcagggc agagccatct attgcttaca
     1501 tttgcttcta gcctgcaagt cgaggagcgc agccttccag aagcagagcg cggcgcctta
     1561 agctgcagaa gttggtcgtg aggcactggg cacgtaagta tcaaggttac aagacaggtt
     1621 taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag
     1681 gcacctattg gtcttactga catccacttt gcctttctct ccacagcgag ctcgctagcc
     1741 accatgctcg aagtcaagga agcatcacca accagcatcc agatcagctg ggtgctccac
     1801 ttgcgccacg ttcgctacta ccgcatcacc tacggtgaaa ctggtggcaa tagccctgtc
     1861 caggaattca ccgtgcctgg cagcaagtcc actgctacca tcagcggcct gaaacctggt
     1921 gtcgactata ccatcacggt gtacgccgtc acgatcttca gcgcctaccg ctccgcctgg
     1981 ccgccgatct ccatcaacta ccgcaccgga accgattaca aggatgacga cgataagggt
     2041 agcggctcca gtagatctgg gcaattggtg agcaagggcg aggagctgtt caccggggtg
     2101 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtccgcggc
     2161 gagggcgagg gcgatgccac caacggcaag ctgaccctga agttcatctg caccaccggc
     2221 aagctgcccg tgccctggcc caccctcgtg accaccttag gctacggcgt ggcctgcttc
     2281 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     2341 tacgtccagg agcgcaccat ctctttcaag gacgacggta cctacaagac ccgcgccgag
     2401 gtgaagttcg agggcgacac cctggtgaac cgcatcgtgc tgaagggcat cgacttcaag
     2461 gaggacggca acatcctggg gcacaagctg gagtacaact tcaacagcca caaggtctat
     2521 atcacggccg acaagcagaa gaacggcatc aaggctaact tcaagacccg ccacaacgtt
     2581 gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     2641 cccgtgctgc tgcccgacaa ccactacctg agccatcagt ccaaactgag caaagacccc
     2701 aacgagaagc gcgatcacat ggtcctgaag gagagggtga ccgccgccgg gattacacat
     2761 gacatggacg agctgtacaa gactagttct aggggagctg gcgctggagc gggtgcaggg
     2821 gctggctcta gattccagtg ccggatctgc atgcggaact tcagcgaccg gtccaacctg
     2881 agcaggcaca tcagaaccca caccggagaa aagcccttcg cctgcgacat ttgcggccgg
     2941 aagttcgcca tcagcagcaa cctgaacagc cacaccaaga tccacactgg cagccagaaa
     3001 cctttccagt gcagaatttg tatgagaaac tttagcagaa gcgacaacct ggccagacac
     3061 atccggacac atactggtga aaaacctttt gcctgtgata tctgtggcag aaagtttgcc
     3121 acctccggca atctgacccg gcacacaaag attcacctgc ggggcagcca gctatcgatt
     3181 gtcgacgctc ctgaacaacg tgaaggtgct tctcaagttt ctgtttctgt tacttttgaa
     3241 gatgttgctg ttctttttac tcgtgatgaa tggaaaaaac ttgatctttc tcaacgttct
     3301 ctttatcgtg aagttatgct tgaaaattat tctaatcttg cttctatggc ttaaggcgcg
     3361 ccccacccct gcagggaatt cgatatcaag cttatcgata atcaacctct ggattacaaa
     3421 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3481 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3541 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     3601 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     3661 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3721 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3781 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt
     3841 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     3901 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     3961 cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctgtg
     4021 atagcggcca tcaagctggc cgcgactcta gatcataatc agccatacca catttgtaga
     4081 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     4141 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4201 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4261 actcatcaat gtatcagctt atcgataccg catgcacgtg cggaccgagc ggccgcagga
     4321 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     4381 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     4441 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4501 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     4561 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     4621 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4681 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4741 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4801 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4861 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     4921 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     4981 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     5041 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     5101 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     5161 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     5221 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5281 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5341 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5401 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5461 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5521 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5581 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5641 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5701 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5761 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5821 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5881 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5941 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     6001 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     6061 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6121 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6181 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6241 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6301 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6361 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttc
//
