LOCUS       sequence-365037-        5641 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  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-sDarken
FEATURES             Location/Qualifiers
     source          1..5641
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        83..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1543..1564
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1670..1691)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1931..1950)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2451..2572
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2488..2507)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2542..2561
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2579..3034)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2716..2737)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2928..2947
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3061..3165
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3167..3524
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3167..3358
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3190..3210)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3375..3510
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3559..4353
                     /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             3559..4353
                     /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(3613..3632)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4223..4242
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4541..4560)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4585..4632
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4961..5549
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5450..5469
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
      601 accgtcagat ccgctagcgc taccggactc agatctcgag ctcaagctta tggatgtgct
      661 cagccctggt cagggcaaca acaccacatc accaccggct ccctttgaga ccggcggcaa
      721 cactactggt atctccgacg tgaccgtcag ctaccaagtg atcacctctc tgctgctggg
      781 cacgctcatc ttctgcgcgg tgctgggcaa tgcgtgcgtg gtggctgcca tcgccttgga
      841 gcgctccctg cagaacgtgg ccaattatct tattggctct ttggcggtca ccgacctcat
      901 ggtgtcggtg ttggtgctgc ccatggccgc gctgtatcag gtgctcaaca agtggacact
      961 gggccaggta acctgcgacc tgttcatcgc cctcgacgtg ctgtgctgca cctcatccat
     1021 cttgcacctg tgcgccatcg cgctggacag gtactgggcc atcacggacc ccatcgacta
     1081 cgtgaacaag aggacgcccc ggcgcgccgc tgcgctcatc tcgctcactt ggcttattgg
     1141 cttcctcatc tctatcccgc ccatgctggg ctggcgcacc ccggaagacc gctcggaccc
     1201 cgacgcatgc accattagca aggatcatgg ctacactatc tattccacct ttggagcttt
     1261 ctacatcccg ctgctgctca tgctggttct ctatgggcgc atattccgag ctgcgcgctt
     1321 cctgagctca tataagaacg tctatatcaa ggccgacaag cagaagaacg gcatcaaggc
     1381 gaacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgcct accactacca
     1441 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcgt
     1501 gcagtccaaa ctttcgaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1561 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggcg gtaccggagg
     1621 gagcatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1681 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggtgagg gcgatgccac
     1741 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1801 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     1861 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacatccagg agcgcaccat
     1921 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1981 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2041 gcacaagctg gagtacaact gttttgacca actgcgagag aggaagacag tgaagacgct
     2101 gggcatcatc atgggcacct tcatcctctg ctggctgccc ttcttcatcg tggctcttgt
     2161 tctgcccttc tgcgagagca gctgccacat gcccaccctg ttgggcgcca taatcaattg
     2221 gctgggctac tccaactctc tgcttaaccc cgtcatttac gcatacttca acaaggactt
     2281 tcaaaacgcg tttaagaaga tcattaagtg taagttctgc cgccagtgag cggccgcgac
     2341 tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     2401 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     2461 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     2521 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa
     2581 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     2641 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     2701 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     2761 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     2821 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     2881 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     2941 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     3001 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     3061 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3121 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     3181 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3241 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3301 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3361 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3421 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     3481 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     3541 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     3601 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     3661 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     3721 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     3781 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     3841 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     3901 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     3961 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4021 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4081 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4141 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4201 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4261 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4321 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4381 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4441 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4501 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     4561 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     4621 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     4681 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     4741 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     4801 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4861 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4921 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4981 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5041 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5101 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5161 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5221 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5281 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5341 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5401 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5461 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5521 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5581 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     5641 a
//
