LOCUS       sequence-260219-        4452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for serotonin (bacterial expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4452)
  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 4452)
  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: pRSET-iSeroSnFR
FEATURES             Location/Qualifiers
     source          1..4452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             112..129
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             133..165
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(169..177,178..192)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     651..672
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1853..1871)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1867..1914
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2011..2466
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2098..2117)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2308..2329
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2492..2596
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2597..2665,2666..3457)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTAGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2815..2834)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3628..4216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4117..4136
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4370..4387
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttc tcatcatcat
      121 catcatcatg gtatggcaag catgactggt ggacagcaaa tgggtcggga tctgtacgac
      181 gatgacgata aggatcgatg gggatccgcg aacgacaccg tagttgtggg ctcgattaac
      241 catacagaac agattatcgt cgcaaacatg ttggcagaga tgattgaggc gcatacagac
      301 cttaaggtgg ttcgcaaact gaaccttggc ggggttaacg ttaactttga agccattaaa
      361 cgcggaggtg cgaataatgg tattgacatt tacctggagt acgttgggta tggtcttgtg
      421 gatattctgg ggtacccgga gccgaacgtc tatatcatcg ccgacaagca gaagaacggc
      481 atcaaggcga acttcaagat ccgctacaac gtggaggacg gcagcgtgca gctcgccgac
      541 cactaccagc agaacacccc catcggcgac ggccctgtgc tgctgcctga caaccactac
      601 ctgagcaccc agtccgtgct gagcaaagac cccaacgaga agcgcgatca catggtcctg
      661 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggcggt
      721 accggaggga gcatgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
      781 gagctggacg gcggcgtaaa cggccacaag ttcagcgtgc gcggcgaggg cgagggcgat
      841 gccaccaacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
      901 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
      961 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1021 accatcagct tcaaggacga cggcacctac aagacccgcg ccgaggtgaa gttcgagggc
     1081 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1141 ctggggcaca agctggagta caactttccg ccgcccggtg ctactgatcc agaaggtgca
     1201 tacgaaaccg tgaagaagga gtacaaacgt aaatggaata ttgtatggct caaaccactg
     1261 ggattcaacg cttcgtatgt gcttgcggtt aaagacgaac tggcgaaaca gtataacctt
     1321 aaaaccttca gtgacttagc gaaaatctcg gataagctga ttctgggtgc aaacatgatg
     1381 tttttagaaa atcccgatgg ttacccaggc ctgcaaaaac tgtacaattt caaattcaag
     1441 cacaccaaaa gcatggacgc tggtattccg tataccgcca ttgataataa cgaagttcag
     1501 gtaattgatg ccactgccac tgatggcttg ctggtgagcc acaaattaaa aattctggag
     1561 gatgataaag cgttcttccc gccgtattat gctgccccca tcatccgtca ggatgtctta
     1621 gataagcatc ctgaactgaa ggacgtgctg aacaaactcg cgaatcaaat ttcactggaa
     1681 gaaatgcaga aactgaatta caagcgtgac ggtgagggtc aggacccagc gaaagtagct
     1741 aaggagtttt tgaaagagaa aggtttaatt taactgcagc tggtaccatg gaattcgaag
     1801 cttgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     1861 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     1921 ggaggaacta tatccggatc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     1981 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     2041 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2101 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2161 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2221 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     2281 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     2341 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     2401 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     2461 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2521 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2581 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     2641 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     2701 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     2761 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     2821 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     2881 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     2941 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3001 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3061 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3121 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     3181 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     3241 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     3301 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     3361 tatcgtagtt atctacacgg cggggagtca ggcaactatg gatgaacgaa atagacagat
     3421 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3481 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3541 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3601 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3661 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3721 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3781 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3841 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3901 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3961 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4021 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4081 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4141 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4201 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4261 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     4321 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     4381 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     4441 tcattaatgc ag
//
