LOCUS Exported 5419 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Peroxide-insensitive version of HyPer to be used as a pH-control. Can be applied as a pH sensor.. ACCESSION . VERSION . KEYWORDS pC1-HyPer-C199S SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5419) AUTHORS Belousov VV, Fradkov AF, Lukyanov KA, Staroverov DB, Shakhbazov KS, Terskikh AV, Lukyanov S TITLE Genetically encoded fluorescent indicator for intracellular hydrogen peroxide. JOURNAL Nat Methods. 2006 Apr;3(4):281-6. PUBMED 16554833 REFERENCE 2 (bases 1 to 5419) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5419 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 597..606 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 603..2039 /codon_start=1 /product="improved fluorescent indicator for intracellular H2O2 (Bilan et al., 2013)" /label=HyPer-3 /translation="MEMASQQGETMSGPLHIGLIPTVGPYLLPHIIPMLHQTFPKLEMY LHEAQTHQLLAQLDSGKLDCVILALVKESEAFIEVPLFDEPMLLAIYEDHPWANRECVP MADLAGEKLLMLEDGHSLRDQAMSAGYNSDNVYIMADKQKNGIKANFKIRHNVEDGSVQ LADHYQQNTPIGDGPVLLPDNHYLSFQSVLSKDPNEKRDHMVLLEFVTAAGITLGMDEL YNVDGGSGGTGSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKLICT TGKLPVPWPTLVTTLGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKT RAEVKFEGDTLVNRIELKGIGFKEDGNILGHKLEYNGTGFCFEAGADEDTHFRATSLET LRNMVAAGSGITLLPALAVPPERKRDGVVYLPCIKPEPRRTIGLVYRPGSPLRSRYEQL AEAIRARMDGHFDKVLKQAV" primer_bind 1211..1232 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1341..1362) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1602..1621) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" polyA_signal 2207..2328 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2244..2263) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2298..2317 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2335..2790) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2472..2493) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2684..2703 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2817..2921 /gene="bla" /label=AmpR promoter promoter 2923..3280 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2946..2966) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3131..3266 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3193..3212 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3315..4109 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3369..3388) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3979..3998 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4297..4316) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4341..4388 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4717..5305 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5206..5225 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcca 601 ccatggagat ggcaagccag cagggcgaga cgatgtccgg accgctgcac attggtttga 661 ttcccacagt tggaccgtac ctgctaccgc atattatccc tatgctgcac cagacctttc 721 caaagctgga aatgtatctg catgaagcac agacccacca gttactggcg caactggaca 781 gcggcaaact cgattgcgtg atcctcgcgc tggtgaaaga gagcgaagca ttcattgaag 841 tgccgttgtt tgatgagcca atgttgctgg ctatctatga agatcacccg tgggcgaacc 901 gcgaatgcgt accgatggcc gatctggcag gggaaaaact gctgatgctg gaagatggtc 961 actctttgcg cgatcaggca atgtccgccg gctacaacag cgacaacgtc tatatcatgg 1021 ccgacaagca gaagaacggc atcaaggcca acttcaagat ccgccacaac gtcgaggacg 1081 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc 1141 tgctgcccga caaccactac ctgagcttcc agtccgtcct gagcaaagac cccaacgaga 1201 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg 1261 acgagctgta caacgtggat ggcggtagcg gtggcaccgg cagcaagggc gaggagctgt 1321 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca 1381 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct 1441 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctc ggctacggcc 1501 tgaagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca 1561 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga 1621 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca 1681 tcggcttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac ggcaccggtt 1741 tctgttttga agccggggcg gatgaagata cacacttccg cgcgaccagc ctggaaactc 1801 tgcgcaacat ggtggcggca ggtagcggga tcactttact gccagcgctg gctgtgccgc 1861 cggagcgcaa acgcgatggg gttgtttatc tgccgtgcat taagccggaa ccacgccgca 1921 ctattggcct ggtttatcgt cctggctcac cgctgcgcag ccgctatgag cagctggcag 1981 aggccatccg cgcaagaatg gatggccatt tcgataaagt tttaaaacag gcggtttaaa 2041 agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccaccggat ctagataact 2101 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca 2161 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc 2221 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt 2281 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg 2341 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta 2401 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt 2461 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca 2521 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa 2581 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat 2641 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag 2701 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg 2761 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg 2821 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat 2881 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca 2941 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag 3001 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc 3061 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct 3121 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg 3181 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa 3241 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat 3301 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg 3361 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg 3421 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg 3481 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc 3541 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg 3601 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca 3661 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc 3721 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg 3781 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg 3841 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata 3901 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg 3961 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat 4021 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct 4081 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca 4141 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt 4201 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat 4261 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc 4321 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc 4381 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag 4441 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc 4501 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg 4561 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt 4621 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg 4681 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt 4741 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca 4801 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac 4861 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac 4921 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct 4981 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg 5041 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca 5101 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt 5161 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta 5221 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc 5281 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc 5341 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa 5401 ccgtattacc gccatgcat //