LOCUS       sequence-392984-        6436 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetically encoded fluorescent probe for labelling endogenous 
            excitatory synapses in vivo.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6436)
  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 6436)
  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-ZnF-hSyn-DIO-PSD95FingR-mGreenLantern
FEATURES             Location/Qualifiers
     source          1..6436
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(62..650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(142..161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(821..1612,1613..1681))
                     /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     1444..1463
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1682..1786)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1854..1872
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1910..1932)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2032..2051
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2068..2523)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2205..2226)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2417..2436
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2598..2738
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2609..2738
                     /label=AAV2 ITR
     polyA_signal    2778..3254
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     3036..3055
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    3286..3874
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3392..3403
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3801..3821
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    complement(3899..3932)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3983..4016)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             4297..4320
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(4351..4353,4354..5064)
                     /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(4393..4414)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             5116..5457
                     /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             5506..5631
                     /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"
     protein_bind    5646..5679
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5730..5763
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     promoter        complement(5801..6248)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    6278..6289
                     /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)"
     repeat_region   6307..6436
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
       61 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      121 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      181 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      241 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      301 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      361 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      421 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      481 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      541 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      601 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      661 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      721 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      781 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      841 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
      901 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
      961 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1021 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1081 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1141 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1201 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1261 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1321 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1381 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1441 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1501 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1561 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1621 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1681 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1741 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1801 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     1861 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     1921 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     1981 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     2041 agcagattgt actgagagtg caccataaaa ttgtaaacgt taatattttg ttaaaattcg
     2101 cgttaaattt ttgttaaatc agctcatttt ttaaccaata gaccgaaatc ggcaaaatcc
     2161 cttataaatc aaaagaatag cccgagatag agttgagtgt tgttccagtt tggaacaaga
     2221 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     2281 atggcccact acgtgaacca tcacccaaat caagtttttt ggggtcgagg tgccgtaaag
     2341 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     2401 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctaaggcg ctggcaagtg
     2461 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     2521 cgtactatgg ttgctttgac gtatgcggtg tgaaataccg cacagatgcg taaggagaaa
     2581 ataccgcatc aggcgcccct gcaggcagct gcgcgctcgc tcgctcactg aggccgcccg
     2641 ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg
     2701 cagagaggga gtggccaact ccatcactag gggttcctgc ggccgctcgg tccgcacgtg
     2761 gttacctaca aaatcagaag gacagggaag ggagcagtgg ttcacgcctg taatcccagc
     2821 aatttgggag gccaaggtgg gtagatcacc tgagattagg agttggagac cagcctggcc
     2881 aatatggtga aaccccgtct ctaccaaaaa aacaaaaatt agctgagcct ggtcatgcat
     2941 gcctggaatc ccaacaactc gggaggctga ggcaggagaa tcgcttgaac ccaggaggcg
     3001 gagattgcag tgagccaaga ttgtgccact gcactccagc ttggttccca atagaccccg
     3061 caggccctac aggttgtctt cccaacttgc cccttgctcc ataccacccc cctccacccc
     3121 ataatattat agaaggacac ctagtcagac aaaatgatgc aacttaattt tattaggaca
     3181 aggctggtgg gcactggagt ggcaacttcc agggccagga gaggcactgg ggaggggtca
     3241 cagggatgcc acccgtagat ctctcgagca gcgctcggta tcgatgcggg gaggcggccc
     3301 aaagggagat ccgactcgtc tgagggcgaa ggcgaagacg cggaagaggc cgcagagccg
     3361 gcagcaggcc gcgggaagga aggtccgctg gattgagggc cgaagggacg tagcagaagg
     3421 acgtcccgcg cagaatccag gtggcaacat aggcgagcag ccaaggaaag gacgatgatt
     3481 tccccgacaa caccacggaa ttgtcagtgc ccaacagccg agcccctgtc cagcagcggg
     3541 caaggcaggc ggcgatgagt tccgccgtgg caatagggag ggggaaagcg aaagtcccgg
     3601 aaaggagctg acaggtggtg gcaatgcccc aaccagtggg ggttgcgtca gcaaacacag
     3661 tgcacaccac gccacgttgc ctgacaacgg gccacaactc ctcataaaga gacagcaacc
     3721 aggatttata caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag
     3781 gcattaaagc agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca
     3841 atctttcaca aattttgtaa tccagaggtt gattatcgat aagcttgata tcgaattcat
     3901 aacttcgtat agcatacatt atacgaagtt attctttgca ccattctaaa gaataacagt
     3961 gataatttct gggttaaggc aaataacttc gtataggata ctttatacga agttatgcta
     4021 gccaccatgc tcgaagtcaa ggaagcatca ccaaccagca tccagatcag ctgggtgctc
     4081 cacttgcgcc acgttcgcta ctaccgcatc acctacggtg aaactggtgg caatagccct
     4141 gtccaggaat tcaccgtgcc tggcagcaag tccactgcta ccatcagcgg cctgaaacct
     4201 ggtgtcgact ataccatcac ggtgtacgcc gtcacgatct tcagcgccta ccgctccgcc
     4261 tggccgccga tctccatcaa ctaccgcacc ggaaccgatt acaaggatga cgacgataag
     4321 ggtagcggct ccagtagatc tgggcaattg gtgagcaagg gcgaggagct gttcaccggg
     4381 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtccgc
     4441 ggcgagggcg agggcgatgc caccaacggc aagctgaccc tgaagttcat ctgcaccacc
     4501 ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct taggctacgg cgtggcctgc
     4561 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     4621 ggctacgtcc aggagcgcac catctctttc aaggacgacg gtacctacaa gacccgcgcc
     4681 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg tgctgaaggg catcgacttc
     4741 aaggaggacg gcaacatcct ggggcacaag ctggagtaca acttcaacag ccacaaggtc
     4801 tatatcacgg ccgacaagca gaagaacggc atcaaggcta acttcaagac ccgccacaac
     4861 gttgaggacg gcggcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     4921 ggccccgtgc tgctgcccga caaccactac ctgagccatc agtccaaact gagcaaagac
     4981 cccaacgaga agcgcgatca catggtcctg aaggagaggg tgaccgccgc cgggattaca
     5041 catgacatgg acgagctgta caagactagt tctaggggag ctggcgctgg agcgggtgca
     5101 ggggctggct ctagattcca gtgccggatc tgcatgcgga acttcagcga ccggtccaac
     5161 ctgagcaggc acatcagaac ccacaccgga gaaaagccct tcgcctgcga catttgcggc
     5221 cggaagttcg ccatcagcag caacctgaac agccacacca agatccacac tggcagccag
     5281 aaacctttcc agtgcagaat ttgtatgaga aactttagca gaagcgacaa cctggccaga
     5341 cacatccgga cacatactgg tgaaaaacct tttgcctgtg atatctgtgg cagaaagttt
     5401 gccacctccg gcaatctgac ccggcacaca aagattcacc tgcggggcag ccagctatcg
     5461 attgtcgacg ctcctgaaca acgtgaaggt gcttctcaag tttctgtttc tgttactttt
     5521 gaagatgttg ctgttctttt tactcgtgat gaatggaaaa aacttgatct ttctcaacgt
     5581 tctctttatc gtgaagttat gcttgaaaat tattctaatc ttgcttctat ggcttaaggc
     5641 gcgccataac ttcgtataat gtatgctata cgaagttatt aagaggtttc atattgctaa
     5701 tagcagctac aatccagcta ccattctgca taacttcgta taaagtatcc tatacgaagt
     5761 tattccggag tcgactctag aggatccggt accttctcga ctgcgctctc aggcacgaca
     5821 cgactcctcc gctgcccacc gcagactgag gcagcgctga gtcgccggcg ccgcagcgca
     5881 gatggtcgcg cccgtgcccc cctatctcgc gcctcgcgtg gtgcggtccg gctgggccgg
     5941 cggcggcgcg gacgcgacca aggtggccgg gaaggggagt ttgcggggga ccggcgagtg
     6001 acgtcagcgc gccttcagtg ctgaggcggc ggtggcgcgc gccgccaggc gggggcgaag
     6061 gcactgtccg cggtgctgaa gctggcagtg cgcacgcgcc tcgccgcatc ctgtttcccc
     6121 tccccctctc tgatagggga tgcgcaattt ggggaatggg ggttgggtgc ttgtccagtg
     6181 ggtcggggtc ggtcgtcagg taggcacccc caccccgcct catcctggtc ctaaaaccca
     6241 cttgcactca tacgcagggc cctctgcagt ctaggacgat gaggatgact ctacgcgtgc
     6301 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     6361 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag cgagcgagcg
     6421 cgcagctgcc tgcagg
//
