LOCUS       sequence-383224-        6048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High affinity version of the 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-H-sDarken
FEATURES             Location/Qualifiers
     source          1..6048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     162..181
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   340..469
                     /label=AAV2 ITR
     promoter        569..1016
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      1029..1038
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1928..1949
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2055..2076)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    2739..3327
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2792..2812)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3210..3221)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3368..3575
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3745..3874
                     /label=AAV2 ITR
     rep_origin      complement(4111..4699)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4191..4210)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4870..5661,5662..5730))
                     /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     5493..5512
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5731..5835)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(5861..6048,1..268))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5998..6019)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
       61 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
      121 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
      181 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
      241 gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg
      301 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tgcgcgctcg ctcgctcact
      361 gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc
      421 gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctt gtagttaatg
      481 attaacccgc catgctactt atctacgtag ccatgctcta ggaagatcgt aggttaatta
      541 atctagactg cagagggccc tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc
      601 ggggtggggg tgcctacctg acgaccgacc ccgacccact ggacaagcac ccaaccccca
      661 ttccccaaat tgcgcatccc ctatcagaga gggggagggg aaacaggatg cggcgaggcg
      721 cgtgcgcact gccagcttca gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc
      781 gccaccgccg cctcagcact gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc
      841 cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg
      901 cgaggcgcga gatagggggg cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag
      961 cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcga
     1021 gaggatccgc caccatggat gtgctcagcc ctggtcaggg caacaacacc acatcaccac
     1081 cggctccctt tgagaccggc ggcaacacta ctggtatctc cgacgtgacc gtcagctacc
     1141 aagtgatcac ctctctgctg ctgggcacgc tcatcttctg cgcggtgctg ggcaatgcgt
     1201 gcgtggtggc tgccatcgcc ttggagcgct ccctgcagaa cgtggccaat tatcttattg
     1261 gctctttggc ggtcaccgac ctcatggtgt cggtgttggt gctgcccatg gccgcgctgt
     1321 atcaggtgct caacaagtgg acactgggcc aggtaacctg cgacctgttc atcgccctcg
     1381 acgtgctgtg ctgcacctca tccatcttgc acctgtgcgc catcgcgctg gacaggtact
     1441 gggccatcac ggaccccatc gactacgtga acaagaggac gccccggcgc gccgctgcgc
     1501 tcatctcgct cacttggctt attggcttcc tcatctctat cccgcccatg ctgggctggc
     1561 gcaccccgga agaccgctcg gaccccgacg catgcaccat tagcaaggat catggctaca
     1621 ctatctattc cacctttgga gctttctaca tcccgctgct gctcatgctg gttctctatg
     1681 ggcgcatatt ccgagctgcg cgcttcctga gctcatataa gaacgtctat atcaccgccg
     1741 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacgtg gaggacggca
     1801 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1861 tgcccgacaa ccactacctg agcacccagt ccgtgctgag caaagacccc aacgagaagc
     1921 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactcac ggcatggacg
     1981 agctgtacaa gggcggtacc ggagggagca tggtgagcaa gggcgaggag ctgttcaccg
     2041 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgc
     2101 gcggcgaggg cgagggcgat gccaccaacg gcaagctgac cctgaagttc atctgcacca
     2161 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     2221 gcttcagccg ctaccccgac cacatgaagc gccacgactt cttcaagtcc gccatgcccg
     2281 aaggctacgt ccaggagcgc accatcagct tcaaggacga cggcacctac aagacccgcg
     2341 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     2401 tcaaggagga cggcaacatc ctggggcaca agctggagta caactgtttt gaccaactgc
     2461 gagagaggaa gacagtgaag acgctgggca tcatcatggg caccttcatc ctctgctggc
     2521 tgcccttctt catcgtggct cttgttctgc ccttctgcga gagcagctgc cacatgccca
     2581 ccctgttggg cgccataatc aattggctgg gctactccaa ctctctgctt aaccccgtca
     2641 tttacgcata cttcaacaag gactttcaaa acgcgtttaa gaagatcatt aagtgtaagt
     2701 tctgccgcca gtgagaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa
     2761 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     2821 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     2881 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     2941 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     3001 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     3061 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     3121 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     3181 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     3241 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     3301 ggatctccct ttgggccgcc tccccgcatc gataccgtcg acccgggcgg ccgctcataa
     3361 agctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3421 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3481 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3541 gaggattggg aagacaatag caggcatgct ggggatgcgg tccggactgt acaattgcat
     3601 tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct
     3661 ctacaaatgt ggtaaaatcg ataaggatct tcctagagca tggctacgta gataagtagc
     3721 atggcgggtt aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc
     3781 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     3841 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcat taatgaatcg gccaacgcgc
     3901 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3961 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4021 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4081 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4141 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4201 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4261 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4321 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4381 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4441 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4501 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4561 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4621 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4681 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4741 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4801 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4861 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4921 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4981 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5041 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5101 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5161 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5221 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5281 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5341 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5401 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5461 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5521 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5581 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5641 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5701 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5761 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5821 aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt
     5881 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     5941 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     6001 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaa
//
