LOCUS Exported 4452 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Fluorescent reporter for serotonin (bacterial expression). ACCESSION . VERSION . KEYWORDS pRSET-iSeroSnFR 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4452 /organism="synthetic DNA construct" /mol_type="other DNA" 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 169..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 2597..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 //