LOCUS       sequence-422889-        8134 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent biosensor for intracellular serotonin (5-HT).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8134)
  AUTHORS   Bukowski L, Strøm M, Andersen J, Maesen J, Tian L, Sinning S
  TITLE     5-HT_FAsTR: A versatile, label-free, microplate assay to quantify 
            serotonin transport and release using the genetically encoded 
            fluorescent biosensor iSeroSnFR
  JOURNAL   Research Square PREPRINT. 18 August 2023. rs.3.rs-3272320/v1
REFERENCE   2  (bases 1 to 8134)
  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: "Research 
            Square PREPRINT. 18 August 2023. rs.3.rs-3272320/v1"
COMMENT     Sequence Label: pIRES_iSeroSnFR_cytosolic
FEATURES             Location/Qualifiers
     source          1..8134
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1558..1579
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2690..2719
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    join(2802..3376,3377..3379,3380..3388)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2969..2986)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3196..3215
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3393..3791
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3838..3959
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3849..3868)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3903..3922
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4145..4600
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4232..4251)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4442..4463
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4717..5074
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4717..4908
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4740..4760)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4925..5060
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5125..5919
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5179..5198)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5789..5808
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5983..6031
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6072..6091)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6191..6213
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6251..6269)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6337..6441
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6442..6510,6511..7302)
                     /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     complement(6660..6679)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7473..8061
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7962..7981
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc ctcgatagcc tcgagggatc catggcgaac gacaccgtag ttgtgggctc
     1141 gattaaccat acagaacaga ttatcgtcgc aaacatgttg gcagagatga ttgaggcgca
     1201 tacagacctt aaggtggttc gcaaactgaa ccttggcggg gttaacgtta actttgaagc
     1261 cattaaacgc ggaggtgcga ataatggtat tgacatttac ctggagtacg ttgggtatgg
     1321 tcttgtggat attctggggt acccggagcc gaacgtctat atcatcgccg acaagcagaa
     1381 gaacggcatc aaggcgaact tcaagatccg ctacaacgtg gaggacggca gcgtgcagct
     1441 cgccgaccac taccagcaga acacccccat cggcgacggc cctgtgctgc tgcctgacaa
     1501 ccactacctg agcacccagt ccgtgctgag caaagacccc aacgagaagc gcgatcacat
     1561 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1621 gggcggtacc ggagggagca tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     1681 cctggtcgag ctggacggcg gcgtaaacgg ccacaagttc agcgtgcgcg gcgagggcga
     1741 gggcgatgcc accaacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     1801 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     1861 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     1921 ggagcgcacc atcagcttca aggacgacgg cacctacaag acccgcgccg aggtgaagtt
     1981 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     2041 caacatcctg gggcacaagc tggagtacaa ctttccgccg cccggtgcta ctgatccaga
     2101 aggtgcatac gaaaccgtga agaaggagta caaacgtaaa tggaatattg tatggctcaa
     2161 accactggga ttcaacgctt cgtatgtgct tgcggttaaa gacgaactgg cgaaacagta
     2221 taaccttaaa accttcagtg acttagcgaa aatctcggat aagctgattc tgggtgcaaa
     2281 catgatgttt ttagaaaatc ccgatggtta cccaggcctg caaaaactgt acaatttcaa
     2341 attcaagcac accaaaagca tggacgctgg tattccgtat accgccattg ataataacga
     2401 agttcaggta attgatgcca ctgccactga tggcttgctg gtgagccaca aattaaaaat
     2461 tctggaggat gataaagcgt tcttcccgcc gtattatgct gcccccatca tccgtcagga
     2521 tgtcttagat aagcatcctg aactgaagga cgtgctgaac aaactcgcga atcaaatttc
     2581 actggaagaa atgcagaaac tgaattacaa gcgtgacggt gagggtcagg acccagcgaa
     2641 agtagctaag gagtttttga aagagaaagg tttaattctg caggtcgacg aacaaaaact
     2701 catctcagaa gaggatctgt aagctctaga attcacgcgt cgctagaatt cacgcgtcga
     2761 gcatgcatct agggcggcca attccgcccc tctccccccc ccccctctcc ctcccccccc
     2821 cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta
     2881 ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc
     2941 ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat
     3001 gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc
     3061 ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt
     3121 gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt
     3181 gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag
     3241 aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt
     3301 tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga
     3361 aaaacacgat gataagcttg ccacaacccg ggatggccaa gcctttgtct caagaagaat
     3421 ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca
     3481 gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc
     3541 attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag
     3601 ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct
     3661 gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg
     3721 acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt
     3781 gggagggcta agcggccgct tccctttagt gagggttaat gcttcgagca gacatgataa
     3841 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     3901 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     3961 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt
     4021 aaagcaagta aaacctctac aaatgtggta aaatccgata aggatcgatc cgggctggcg
     4081 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     4141 atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4201 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4261 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     4321 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4381 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4441 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4501 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4561 tttaacgcga attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc
     4621 ttacgcatct gtgcggtatt tcacaccgca tacgcggatc tgcgcagcac catggcctga
     4681 aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg
     4741 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4801 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4861 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4921 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4981 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     5041 aggaggcttt tttggaggcc taggcttttg caaaaagctt gattcttctg acacaacagt
     5101 ctcgaactta aggctagagc caccatgatt gaacaagatg gattgcacgc aggttctccg
     5161 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5221 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5281 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     5341 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5401 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5461 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5521 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5581 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5641 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5701 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5761 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5821 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5881 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5941 tgaccgacca agcgacgccc aacctgccat cacgatggcc gcaataaaat atctttattt
     6001 tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag cgataaggat ccgcgtatgg
     6061 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     6121 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     6181 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     6241 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     6301 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     6361 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6421 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6481 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6541 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6601 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6661 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6721 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6781 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6841 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6901 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6961 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     7021 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     7081 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     7141 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     7201 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     7261 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     7321 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     7381 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7441 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7501 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7561 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7621 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7681 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7741 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7801 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7861 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7921 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7981 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     8041 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     8101 tgctggcctt ttgctcacat ggctcgacag atct
//
