LOCUS       Exported                5407 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Hydrogen peroxide insensitive version of HyPerRed to be used as a pH
            control in HyPer experiments..
ACCESSION   .
VERSION     .
KEYWORDS    pC1-HyPer-Red-C199S
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5407)
  AUTHORS   Ermakova YG, Bilan DS, Matlashov ME, Mishina NM, Markvicheva KN, 
            Subach OM, Subach FV, Bogeski I, Hoth M, Enikolopov G, Belousov VV
  TITLE     Red fluorescent genetically encoded indicator for intracellular 
            hydrogen peroxide.
  JOURNAL   Nat Commun. 2014 Oct 21;5:5222. doi: 10.1038/ncomms6222.
  PUBMED    25330925
REFERENCE   2  (bases 1 to 5407)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5407
                     /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      598..607
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             604..2031
                     /codon_start=1
                     /product="red fluorescent genetically encoded indicator for
                     H2O2 ()"
                     /label=HyPerRed
                     /translation="MEMASQQGETMSGPLHIGLIPTVGPYLLPHIIPMLHQTFPKLEMY
                     LHEAQTHQLLAQLDSGKLDCVILALVKESEAFIEVPLFDEPMLLAIYEDHPWANRECVP
                     MADLAGEKLLMLEDGHSLRDQAMPRTVSERMYPEDGALKSEIKKGLRLKDGGHYAAEVK
                     TTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQCERAEGRHSTGGMDELYKGGTGGS
                     LVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEAFQTAKLKVTKGGPL
                     PFAWDILSPQFMYGSKAYIKHPADIPDYFKLSFPEGFRWERVMNFEDGGIIHVNQDSSL
                     QDGVFIYKVKLRGTNFPPDGPVMQKKTMGWEAAGVGFCFEAGADEDTHFRATSLETLRN
                     MVAAGSGITLLPALAVPPERKRDGVVYLPCIKPEPRRTIGLVYRPGSPLRSRYEQLAEA
                     IRARMDGHFDKVLKQAV"
     primer_bind     1683..1702
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2195..2316
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2232..2251)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2286..2305
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2323..2778)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2460..2481)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2672..2691
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2805..2909
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2911..3268
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2934..2954)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3119..3254
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3181..3200
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3303..4097
                     /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(3357..3376)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3967..3986
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4285..4304)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4329..4376
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4705..5293
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5194..5213
                     /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 gctagccgcc
      601 accatggaga tggcgagcca gcagggcgag acgatgtccg gaccgctgca cattggtttg
      661 attcccacag ttggaccgta cctgctaccg catattatcc ctatgctgca ccagaccttt
      721 ccaaagctgg aaatgtatct gcatgaagca cagacccacc agttactggc gcaactggac
      781 agcggcaaac tcgattgcgt gatcctcgcg ctggtgaaag agagcgaagc attcattgaa
      841 gtgccgttgt ttgatgagcc aatgttgctg gctatctatg aagatcaccc gtgggcgaac
      901 cgcgaatgcg taccgatggc cgatctggca ggggaaaaac tgctgatgct ggaagatggt
      961 cactctttgc gcgatcaggc aatgccccgc acggtttccg agcggatgta ccccgaggac
     1021 ggcgccctga agagcgagat caagaagggg ctgaggctga aggacggcgg ccactacgcc
     1081 gccgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacatc
     1141 gtcgacatca agttggacat cgtgtcccac aacgaggact acaccatcgt ggaacagtgc
     1201 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gggaggtaca
     1261 ggcgggagtc tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
     1321 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     1381 ggcgagggcc gcccctacga ggcctttcag accgctaagc tgaaggtgac caagggtggc
     1441 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
     1501 attaagcacc cagccgacat ccccgactac ttcaagctgt ccttccccga gggcttcagg
     1561 tgggagcgcg tgatgaactt cgaggacggc ggcattattc acgttaacca ggactcctcc
     1621 ctgcaggacg gcgtattcat ctacaaggtg aagctgcgcg gcaccaactt cccccccgac
     1681 ggccccgtaa tgcagaagaa gaccatgggc tgggaggctg ccggggtggg tttctgtttt
     1741 gaagccgggg cggatgaaga tacacacttc cgcgcgacca gcctggaaac tctgcgcaac
     1801 atggtggcgg caggtagcgg gatcacttta ctgccagcgc tggctgtgcc gccggagcgc
     1861 aaacgcgatg gggttgttta tctgccgtgc attaagccgg aaccacgccg cactattggt
     1921 ctggtttatc gtcctggctc accgctgcgc agccgctatg agcagctggc agaggccatc
     1981 cgcgcaagaa tggatggcca tttcgataaa gttttaaaac aggcggttta atgagaattc
     2041 tgcagtcgac ggtaccgcgg gcccgggatc caccggatct agataactga tcataatcag
     2101 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2161 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2221 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2281 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     2341 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     2401 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     2461 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     2521 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     2581 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     2641 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     2701 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     2761 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     2821 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2881 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     2941 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3001 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3061 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3121 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3181 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3241 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     3301 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3361 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3421 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3481 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     3541 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     3601 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     3661 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     3721 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     3781 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     3841 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     3901 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     3961 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4021 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4081 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4141 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4201 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4261 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     4321 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     4381 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     4441 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     4501 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     4561 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4621 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4681 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4741 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4801 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4861 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4921 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4981 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5041 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5101 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5161 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5221 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5281 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5341 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5401 catgcat
//
