LOCUS       sequence-260416-        6904 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for serotonin (conditional + membrane 
            localization tag).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6904)
  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 6904)
  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
FEATURES             Location/Qualifiers
     source          1..6904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        251..528
                     /label=chicken beta-actin promoter
     primer_bind     807..829
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     891..910
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    945..978
                     /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    1041..1074
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1121..1267)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(1271..1300)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2411..2432)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(2885..2944)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      2941..2950
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2979..3012)
                     /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(3083..3116)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3158..3746
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3211..3231)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3629..3640)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3789..3910
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3800..3819)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3854..3873
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4072..4201
                     /label=AAV2 ITR
     rep_origin      complement(4438..5026)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4518..4537)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5197..5988,5989..6057))
                     /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     5820..5839
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6058..6162)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6188..6643)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6325..6346)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6537..6556
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6715..6844
                     /label=AAV2 ITR
ORIGIN
        1 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
       61 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      121 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      181 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      241 attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
      301 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
      361 gggggggggg ggcgcgcgcc gggggggggg ggggggggsg gggggsgggg sggggcgagg
      421 cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
      481 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg
      541 cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga
      601 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat
      661 tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg
      721 ctccgggagg gccctttgtg cggggggagc ggctcggggc tgtccgcggg gggacggctg
      781 ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag
      841 agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct
      901 ggttattgtg ctgtctcatc attttggcaa agaattggat ccccataact tcgtataaag
      961 tatcctatac gaagttatat caaaatagga agaccaatgc ttcaccatcg acccgaattg
     1021 ccaagcatca ccatcgaccc ataacttcgt ataatgtatg ctatacgaag ttatactagg
     1081 gatcccctta cttgtacaga gctcggcgcg cctcgagcta acgtggcttc ttctgccaaa
     1141 gcatgatgag gatgataagg gagatgatgg tgagcaccac cagggccagg atggctgaga
     1201 tcaccaccac cttaaagggc aaggagtgtg gcaccacgat gacctcctgc gtgtcctggc
     1261 ccacagcatt cagatcctct tctgagatga gtttttgttc gtcgacctgc agaattaaac
     1321 ctttctcttt caaaaactcc ttagctactt tcgctgggtc ctgaccctca ccgtcacgct
     1381 tgtaattcag tttctgcatt tcttccagtg aaatttgatt cgcgagtttg ttcagcacgt
     1441 ccttcagttc aggatgctta tctaagacat cctgacggat gatgggggca gcataatacg
     1501 gcgggaagaa cgctttatca tcctccagaa tttttaattt gtggctcacc agcaagccat
     1561 cagtggcagt ggcatcaatt acctgaactt cgttattatc aatggcggta tacggaatac
     1621 cagcgtccat gcttttggtg tgcttgaatt tgaaattgta cagtttttgc aggcctgggt
     1681 aaccatcggg attttctaaa aacatcatgt ttgcacccag aatcagctta tccgagattt
     1741 tcgctaagtc actgaaggtt ttaaggttat actgtttcgc cagttcgtct ttaaccgcaa
     1801 gcacatacga agcgttgaat cccagtggtt tgagccatac aatattccat ttacgtttgt
     1861 actccttctt cacggtttcg tatgcacctt ctggatcagt agcaccgggc ggcggaaagt
     1921 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     1981 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtaggtgc
     2041 cgtcgtcctt gaagctgatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
     2101 agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca
     2161 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca
     2221 gggtcagctt gccgttggtg gcatcgccct cgccctcgcc gcgcacgctg aacttgtggc
     2281 cgtttacgcc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
     2341 ccttgctcat gctccctccg gtaccgccct tgtacagctc gtccatgccg agagtgatcc
     2401 cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg gggtctttgc
     2461 tcagcacgga ctgggtgctc aggtagtggt tgtcaggcag cagcacaggg ccgtcgccga
     2521 tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcc acgttgtagc
     2581 ggatcttgaa gttcgccttg atgccgttct tctgcttgtc ggcgatgata tagacgttcg
     2641 gctccgggta ccccagaata tccacaagac catacccaac gtactccagg taaatgtcaa
     2701 taccattatt cgcacctccg cgtttaatgg cttcaaagtt aacgttaacc ccgccaaggt
     2761 tcagtttgcg aaccacctta aggtctgtat gcgcctcaat catctctgcc aacatgtttg
     2821 cgacgataat ctgttctgta tggttaatcg agcccacaac tacggtgtcg ttcgcagatc
     2881 tgtcaccagt ggaacctgga acccagagca gcagtaccca tagcaggagt gtgtctgtct
     2941 ccatggtggc actagtgcta gcggtaccga attctagtat aacttcgtat aggatacttt
     3001 atacgaagtt atcattggga ttcttcctat tttgatccaa gcatcaccat cgaccctcta
     3061 gtccagatct caccatcgac ccataacttc gtatagcata cattatacga agttatgtcc
     3121 ctcgaagagg ttcgaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     3181 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     3241 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     3301 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     3361 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     3421 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     3481 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     3541 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     3601 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     3661 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     3721 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cccgggcggc cgcttcgagc
     3781 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     3841 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3901 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     3961 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatcttcc
     4021 tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac aaggaacccc
     4081 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     4141 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     4201 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4261 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4321 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4381 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4441 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4501 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4561 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4621 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4681 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4741 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4801 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4861 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4921 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4981 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5041 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5101 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5161 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5221 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5281 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5341 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5401 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5461 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5521 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5581 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5641 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5701 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5761 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5821 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5881 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5941 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6001 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6061 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6121 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6181 gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     6241 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     6301 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     6361 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     6421 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     6481 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     6541 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     6601 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg
     6661 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctgcgc
     6721 gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc
     6781 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt
     6841 tccttgtagt taatgattaa cccgccatgc tacttatcta cgtagccatg ctctaggaag
     6901 agta
//
