LOCUS       sequence-365012-        5641 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High affinity version of the Serotonin Sensor sDarken under the 
            control of a CMV Promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5641)
  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 5641)
  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: pN1-CMV-H-sDarken
FEATURES             Location/Qualifiers
     source          1..5641
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        75..378
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        379..582
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     533..553
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1535..1556
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1662..1683)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    2443..2564
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2480..2499)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2534..2553
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2571..3026)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2708..2729)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2920..2939
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3053..3157
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3159..3516
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3159..3350
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3182..3202)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3367..3502
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3551..4345
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3551..4345
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3605..3624)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4215..4234
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4533..4552)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4577..4624
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4953..5541
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5442..5461
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
       61 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      121 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      181 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      241 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      301 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      361 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      421 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      481 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      541 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
      601 atccgctagc gctaccggac tcagatctcg agctcaagct tatggatgtg ctcagccctg
      661 gtcagggcaa caacaccaca tcaccaccgg ctccctttga gaccggcggc aacactactg
      721 gtatctccga cgtgaccgtc agctaccaag tgatcacctc tctgctgctg ggcacgctca
      781 tcttctgcgc ggtgctgggc aatgcgtgcg tggtggctgc catcgccttg gagcgctccc
      841 tgcagaacgt ggccaattat cttattggct ctttggcggt caccgacctc atggtgtcgg
      901 tgttggtgct gcccatggcc gcgctgtatc aggtgctcaa caagtggaca ctgggccagg
      961 taacctgcga cctgttcatc gccctcgacg tgctgtgctg cacctcatcc atcttgcacc
     1021 tgtgcgccat cgcgctggac aggtactggg ccatcacgga ccccatcgac tacgtgaaca
     1081 agaggacgcc ccggcgcgcc gctgcgctca tctcgctcac ttggcttatt ggcttcctca
     1141 tctctatccc gcccatgctg ggctggcgca ccccggaaga ccgctcggac cccgacgcat
     1201 gcaccattag caaggatcat ggctacacta tctattccac ctttggagct ttctacatcc
     1261 cgctgctgct catgctggtt ctctatgggc gcatattccg agctgcgcgc ttcctgagct
     1321 catataagaa cgtctatatc accgccgaca agcagaagaa cggcatcaag gccaacttca
     1381 agatccgcca caacgtggag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     1441 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     1501 tgctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1561 ccgccgggat cactcacggc atggacgagc tgtacaaggg cggtaccgga gggagcatgg
     1621 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1681 acgtaaacgg ccacaagttc agcgtgcgcg gcgagggcga gggcgatgcc accaacggca
     1741 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1801 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcgcc
     1861 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcagcttca
     1921 aggacgacgg cacctacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1981 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     2041 tggagtacaa ctgttttgac caactgcgag agaggaagac agtgaagacg ctgggcatca
     2101 tcatgggcac cttcatcctc tgctggctgc ccttcttcat cgtggctctt gttctgccct
     2161 tctgcgagag cagctgccac atgcccaccc tgttgggcgc cataatcaat tggctgggct
     2221 actccaactc tctgcttaac cccgtcattt acgcatactt caacaaggac tttcaaaacg
     2281 cgtttaagaa gatcattaag tgtaagttct gccgccagtg agcggccgcg actctagatc
     2341 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     2401 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     2461 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     2521 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag
     2581 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     2641 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     2701 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     2761 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     2821 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     2881 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     2941 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     3001 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     3061 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3121 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     3181 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3241 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3301 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3361 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3421 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3481 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     3541 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3601 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3661 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3721 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3781 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     3841 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     3901 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     3961 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4021 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     4081 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4141 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4201 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4261 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4321 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4381 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4441 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4501 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     4561 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     4621 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     4681 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     4741 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     4801 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4861 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4921 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4981 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5041 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5101 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5161 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5221 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5281 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5341 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5401 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5461 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5521 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5581 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5641 a
//
