LOCUS       sequence-383222-        6048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Serotonin Sensor sDarken under the control of a hSyn Promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6048)
  AUTHORS   Kubitschke M, Muller M, Wallhorn L, Pulin M, Mittag M, Pollok S, 
            Ziebarth T, Bremshey S, Gerdey J, Claussen KC, Renken K, Gross J, 
            Gneisse P, Meyer N, Wiegert JS, Reiner A, Fuhrmann M, Masseck OA
  TITLE     Next generation genetically encoded fluorescent sensors for 
            serotonin.
  JOURNAL   Nat Commun. 2022 Dec 6;13(1):7525. doi: 10.1038/s41467-022-35200-w.
  PUBMED    36473867
REFERENCE   2  (bases 1 to 6048)
  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: "Nat 
            Commun."; date: "2022-12-6"; volume: "13(1):7525. doi"; pages: " 
            10.1038/s41467-022-35200-w"
COMMENT     Sequence Label: pAAV-SYN-sDarken
FEATURES             Location/Qualifiers
     source          1..6048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     139..158
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   317..446
                     /label=AAV2 ITR
     promoter        546..993
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      1006..1015
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1905..1926
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2032..2053)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2293..2312)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2716..3304
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2769..2789)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3187..3198)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3345..3552
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3722..3851
                     /label=AAV2 ITR
     rep_origin      complement(4088..4676)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4168..4187)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4847..5638,5639..5707))
                     /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     5470..5489
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5708..5812)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(5838..6048,1..245))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5975..5996)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
       61 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
      121 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
      181 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
      241 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
      301 ttcgctatta cgccagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg
      361 gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt
      421 ggccaactcc atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc
      481 tacgtagcca tgctctagga agatcgtagg ttaattaatc tagactgcag agggccctgc
      541 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      601 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      661 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      721 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      781 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      841 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      901 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      961 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagag gatccgccac catggatgtg
     1021 ctcagccctg gtcagggcaa caacaccaca tcaccaccgg ctccctttga gaccggcggc
     1081 aacactactg gtatctccga cgtgaccgtc agctaccaag tgatcacctc tctgctgctg
     1141 ggcacgctca tcttctgcgc ggtgctgggc aatgcgtgcg tggtggctgc catcgccttg
     1201 gagcgctccc tgcagaacgt ggccaattat cttattggct ctttggcggt caccgacctc
     1261 atggtgtcgg tgttggtgct gcccatggcc gcgctgtatc aggtgctcaa caagtggaca
     1321 ctgggccagg taacctgcga cctgttcatc gccctcgacg tgctgtgctg cacctcatcc
     1381 atcttgcacc tgtgcgccat cgcgctggac aggtactggg ccatcacgga ccccatcgac
     1441 tacgtgaaca agaggacgcc ccggcgcgcc gctgcgctca tctcgctcac ttggcttatt
     1501 ggcttcctca tctctatccc gcccatgctg ggctggcgca ccccggaaga ccgctcggac
     1561 cccgacgcat gcaccattag caaggatcat ggctacacta tctattccac ctttggagct
     1621 ttctacatcc cgctgctgct catgctggtt ctctatgggc gcatattccg agctgcgcgc
     1681 ttcctgagct catataagaa cgtctatatc aaggccgaca agcagaagaa cggcatcaag
     1741 gcgaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc ctaccactac
     1801 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1861 gtgcagtcca aactttcgaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1921 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg cggtaccgga
     1981 gggagcatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     2041 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggtga gggcgatgcc
     2101 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     2161 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     2221 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacatcca ggagcgcacc
     2281 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     2341 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     2401 gggcacaagc tggagtacaa ctgttttgac caactgcgag agaggaagac agtgaagacg
     2461 ctgggcatca tcatgggcac cttcatcctc tgctggctgc ccttcttcat cgtggctctt
     2521 gttctgccct tctgcgagag cagctgccac atgcccaccc tgttgggcgc cataatcaat
     2581 tggctgggct actccaactc tctgcttaac cccgtcattt acgcatactt caacaaggac
     2641 tttcaaaacg cgtttaagaa gatcattaag tgtaagttct gccgccagtg agaattcgat
     2701 atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2761 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2821 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     2881 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     2941 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     3001 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     3061 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     3121 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     3181 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     3241 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     3301 ccgcatcgat accgtcgacc cgggcggccg ctcataaagc tcgactgtgc cttctagttg
     3361 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3421 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3481 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3541 gcatgctggg gatgcggtcc ggactgtaca attgcattca ttttatgttt caggttcagg
     3601 gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata
     3661 aggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac
     3721 aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     3781 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     3841 cgagcgcgca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3901 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3961 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4021 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4081 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4141 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4201 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4261 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4321 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4381 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4441 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4501 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4561 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4621 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4681 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4741 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4801 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4861 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4921 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4981 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5041 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5101 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5161 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5221 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5281 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5341 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5401 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5461 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5521 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5581 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5641 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5701 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5761 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5821 cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     5881 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     5941 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     6001 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcaggg
//
