LOCUS Exported 5152 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Fluorescent reporter for serotonin (mammalian expression, membrane localization tag). ACCESSION . VERSION . KEYWORDS pDisplay-iSeroSnFR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5152) 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 5152) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5152 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 9..388 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 389..592 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 543..563 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 637..656 /label=T7 /note="T7 promoter, forward primer" promoter 637..655 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" sig_peptide 736..798 /product="leader sequence from mouse immunoglobulin kappa light chain" /label=Ig-kappa leader primer_bind 1248..1269 /label=EGFP-C /note="EGFP, forward primer" CDS 2380..2409 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2413..2559 /codon_start=1 /product="transmembrane domain from platelet derived growth factor receptor beta" /label=PDGFR-beta TM domain /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ KKPR" primer_bind complement(2581..2598) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2587..2811 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin complement(2943..3531) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3023..3042) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3702..4562) /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 4325..4344 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4563..4667) /gene="bla" /label=AmpR promoter rep_origin 4694..5149 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4781..4800) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4991..5012 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 cgcgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt 61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg 121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc 181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc 241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg 301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat 361 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg 421 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag 481 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt 541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagaa ctctctggct 601 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac 661 ccaagcttgg taccgaactc ggatccacta gtaacggccg ccagtgtgct ggaattcggc 721 ttggggatat ccaccatgga gacagacaca ctcctgctat gggtactgct gctctgggtt 781 ccaggttcca ctggtgacag atctgcgaac gacaccgtag ttgtgggctc gattaaccat 841 acagaacaga ttatcgtcgc aaacatgttg gcagagatga ttgaggcgca tacagacctt 901 aaggtggttc gcaaactgaa ccttggcggg gttaacgtta actttgaagc cattaaacgc 961 ggaggtgcga ataatggtat tgacatttac ctggagtacg ttgggtatgg tcttgtggat 1021 attctggggt acccggagcc gaacgtctat atcatcgccg acaagcagaa gaacggcatc 1081 aaggcgaact tcaagatccg ctacaacgtg gaggacggca gcgtgcagct cgccgaccac 1141 taccagcaga acacccccat cggcgacggc cctgtgctgc tgcctgacaa ccactacctg 1201 agcacccagt ccgtgctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg 1261 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggcggtacc 1321 ggagggagca tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag 1381 ctggacggcg gcgtaaacgg ccacaagttc agcgtgcgcg gcgagggcga gggcgatgcc 1441 accaacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg 1501 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac 1561 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc 1621 atcagcttca aggacgacgg cacctacaag acccgcgccg aggtgaagtt cgagggcgac 1681 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg 1741 gggcacaagc tggagtacaa ctttccgccg cccggtgcta ctgatccaga aggtgcatac 1801 gaaaccgtga agaaggagta caaacgtaaa tggaatattg tatggctcaa accactggga 1861 ttcaacgctt cgtatgtgct tgcggttaaa gacgaactgg cgaaacagta taaccttaaa 1921 accttcagtg acttagcgaa aatctcggat aagctgattc tgggtgcaaa catgatgttt 1981 ttagaaaatc ccgatggtta cccaggcctg caaaaactgt acaatttcaa attcaagcac 2041 accaaaagca tggacgctgg tattccgtat accgccattg ataataacga agttcaggta 2101 attgatgcca ctgccactga tggcttgctg gtgagccaca aattaaaaat tctggaggat 2161 gataaagcgt tcttcccgcc gtattatgct gcccccatca tccgtcagga tgtcttagat 2221 aagcatcctg aactgaagga cgtgctgaac aaactcgcga atcaaatttc actggaagaa 2281 atgcagaaac tgaattacaa gcgtgacggt gagggtcagg acccagcgaa agtagctaag 2341 gagtttttga aagagaaagg tttaattctg caggtcgacg aacaaaaact catctcagaa 2401 gaggatctga atgctgtggg ccaggacacg caggaggtca tcgtggtgcc acactccttg 2461 ccctttaagg tggtggtgat ctcagccatc ctggccctgg tggtgctcac catcatctcc 2521 cttatcatcc tcatcatgct ttggcagaag aagccacgtt aggcggccgc tcgagatcag 2581 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct 2641 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc 2701 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg 2761 aggattggga agacaatagc aggcgtgctg gggatgcggt gggctctatg gcttctgagg 2821 cggaaagaac cagtggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 2881 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 2941 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 3001 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 3061 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 3121 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 3181 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 3241 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 3301 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 3361 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt 3421 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 3481 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 3541 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 3601 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 3661 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 3721 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 3781 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 3841 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 3901 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 3961 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 4021 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 4081 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 4141 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 4201 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 4261 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 4321 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 4381 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 4441 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 4501 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 4561 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 4621 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 4681 aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 4741 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 4801 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 4861 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 4921 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 4981 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 5041 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt 5101 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta cg //