LOCUS       sequence-403406-        6811 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for serotonin imaging (conditional + membrane 
            localization tag with enhanced export sequence).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6811)
  AUTHORS   Unger EK, Keller JP, Altermatt M, Liang R, Matsui A, Dong C, Hon OJ,
            Yao Z, Sun J, Banala S, Flanigan ME, Jaffe DA, Hartanto S, Carlen J,
            Mizuno GO, Borden PM, Shivange AV, Cameron LP, Sinning S, Underhill 
            SM, Olson DE, Amara SG, Temple Lang D, Rudnick G, Marvin JS, Lavis 
            LD, Lester HA, Alvarez VA, Fisher AJ, Prescher JA, Kash TL, 
            Yarov-Yarovoy V, Gradinaru V, Looger LL, Tian L
  TITLE     Directed Evolution of a Selective and Sensitive Serotonin Sensor via
            Machine Learning.
  JOURNAL   Cell. 2020 Dec 12. pii: S0092-8674(20)31612-3. doi: 
            10.1016/j.cell.2020.11.040.
  PUBMED    33333022
REFERENCE   2  (bases 1 to 6811)
  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: "Cell. 2020 
            Dec 12. pii: S0092-8674(20)31612-3. doi: 
            10.1016/j.cell.2020.11.040."
COMMENT     Sequence Label: pAAV-CAG-FLEx-iSeroSnFR-EnhancedExport
FEATURES             Location/Qualifiers
     source          1..6811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1162..1617)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1299..1320)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1511..1530
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1689..1818
                     /label=AAV2 ITR
     promoter        2129..2414
                     /label=chicken beta-actin promoter
     primer_bind     2693..2715
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2777..2796
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    2831..2864
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2927..2960
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(3211..3240)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4345..4366)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(4819..4878)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      4875..4884
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(4913..4946)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(5017..5050)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5092..5680
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5145..5165)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5563..5574)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5723..5844
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5734..5753)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5788..5807
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6223..6811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6303..6322)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     1201 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     1261 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     1321 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     1381 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     1441 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     1501 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     1561 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
     1621 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     1681 tacgccagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg
     1741 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
     1801 ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc
     1861 catgctctag gaagagtacc attgacgtca ataatgacgt atgttcccat agtaacgcca
     1921 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     1981 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     2041 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     2101 tacgtattag tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc
     2161 cccatctccc cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
     2221 tgtgcagcga tgggggcggg gggggggggg ggggggggcg cgcgccaggc ggggcggggc
     2281 ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg
     2341 cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa
     2401 gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg
     2461 cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg
     2521 acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc
     2581 tgtggctgcg tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct
     2641 cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc
     2701 ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt
     2761 ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa
     2821 ttggatcccc ataacttcgt ataaagtatc ctatacgaag ttatatcaaa ataggaagac
     2881 caatgcttca ccatcgaccc gaattgccaa gcatcaccat cgacccataa cttcgtataa
     2941 tgtatgctat acgaagttat actagggatc cccttacttg tacagagctc ggcgcgcctc
     3001 gagctacagg gctacctcat tttcatagca aaaggagtta gcgttctcct ggcgtcgcct
     3061 ttcccgcctc ttctgcaagt atttgatgag gatgataagg gagatgatgg tgagcaccac
     3121 cagggccagg atggctgaga tcaccaccac cttaaagggc aaggagtgtg gcaccacgat
     3181 gacctcctgc gtgtcctggc ccacagcatt cagatcctct tctgagatga gtttttgttc
     3241 acgcgtaatt aaacctttct ctttcaaaaa ctccttagct actttcgctg ggtcctgacc
     3301 ctcaccgtca cgcttgtaat tcagtttctg catttcttcc agtgaaattt gattcgcgag
     3361 tttgttcagc acgtccttca gttcaggatg cttatctaag acatcctgac ggatgatggg
     3421 ggcagcataa tacggcggga agaacgcttt atcatcctcc agaattttta atttgtggct
     3481 caccagcaag ccatcagtgg cagtggcatc aattacctga acttcgttat tatcaatggc
     3541 ggtatacgga ataccagcgt ccatgctttt ggtgtgcttg aatttgaaat tgtacagttt
     3601 ttgcaggcct gggtaaccat cgggattttc taaaaacatc atgtttgcac ccagaatcag
     3661 cttatccgag attttcgcta agtcactgaa ggttttaagg ttatactgtt tcgccagttc
     3721 gtctttaacc gcaagcacat acgaagcgtt gaatcccagt ggtttgagcc atacaatatt
     3781 ccatttacgt ttgtactcct tcttcacggt ttcgtatgca ccttctggat cagtagcacc
     3841 gggcggcgga aagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc
     3901 gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg
     3961 ggtcttgtag gtgccgtcgt ccttgaagct gatggtgcgc tcctggacgt agccttcggg
     4021 catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc tgaagcactg
     4081 cacgccgtag gtcagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt
     4141 gcagatgaac ttcagggtca gcttgccgtt ggtggcatcg ccctcgccct cgccgcgcac
     4201 gctgaacttg tggccgttta cgccgccgtc cagctcgacc aggatgggca ccaccccggt
     4261 gaacagctcc tcgcccttgc tcatgctccc tccggtaccg cccttgtaca gctcgtccat
     4321 gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
     4381 gttggggtct ttgctcagca cggactgggt gctcaggtag tggttgtcag gcagcagcac
     4441 agggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc
     4501 ctccacgttg tagcggatct tgaagttcgc cttgatgccg ttcttctgct tgtcggcgat
     4561 gatatagacg ttcggctccg ggtaccccag aatatccaca agaccatacc caacgtactc
     4621 caggtaaatg tcaataccat tattcgcacc tccgcgttta atggcttcaa agttaacgtt
     4681 aaccccgcca aggttcagtt tgcgaaccac cttaaggtct gtatgcgcct caatcatctc
     4741 tgccaacatg tttgcgacga taatctgttc tgtatggtta atcgagccca caactacggt
     4801 gtcgttcgca gatctgtcac cagtggaacc tggaacccag agcagcagta cccatagcag
     4861 gagtgtgtct gtctccatgg tggcactagt gctagcggta ccgaattcta gtataacttc
     4921 gtataggata ctttatacga agttatcatt gggattcttc ctattttgat ccaagcatca
     4981 ccatcgaccc tctagtccag atctcaccat cgacccataa cttcgtatag catacattat
     5041 acgaagttat gtccctcgaa gaggttcgaa ttcgatatca agcttatcga taatcaacct
     5101 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     5161 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     5221 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     5281 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     5341 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     5401 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     5461 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     5521 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     5581 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     5641 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacccggg
     5701 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     5761 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     5821 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     5881 cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
     5941 gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa
     6001 ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     6061 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6121 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6181 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6241 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6301 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6361 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6421 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6481 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6541 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6601 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6661 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6721 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6781 gcagattacg cgcagaaaaa aaggatctca a
//
