LOCUS       Exported                5419 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pC1-HyPer-3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5419)
  AUTHORS   Bilan DS, Pase L, Joosen L, Gorokhovatsky AY, Ermakova YG, Gadella 
            TW, Grabher C, Schultz C, Lukyanov S, Belousov VV
  TITLE     HyPer-3: a genetically encoded H2O2 probe with improved performance 
            for ratiometric and fluorescence lifetime imaging.
  JOURNAL   ACS Chem Biol. 2012 Dec 20.
  PUBMED    23256573
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"
     primer_bind     complement(54..75)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(315..334)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    920..1041
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(957..976)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1011..1030
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1048..1503)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1185..1206)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1397..1416
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1530..1634
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1636..1993
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(1659..1679)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1844..1979
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1906..1925
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2028..2822
                     /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(2082..2101)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2692..2711
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3010..3029)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3054..3101
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3430..4018
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3919..3938
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4193..4496
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4497..4700
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4651..4671
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4729..4738
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4735..752
                     /codon_start=1
                     /product="improved fluorescent indicator for intracellular 
                     H2O2 (Bilan et al., 2013)"
                     /label=HyPer-3
                     /translation="MEMASQQGETMSGPLHIGLIPTVGPYLLPHIIPMLYQTFPKLEMY
                     LHEAQTHQLLAQLDSGKLDCVILALVKESEAFIEVPLFDEPMLLAIYEDHPWANRECVP
                     MADLAGEKLLMLEDGHCLRDQAMSAGYNSDNVYIMADKQKNGIKANFKIRHNVEDGSVQ
                     LADHYQQNTPIGDGPVLLPDNHYLSFQSVLSKDPNEKRDHMVLLEFVTAAGITLGMDEL
                     YNVDGGSGGTGSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKLICT
                     TGKLPVPWPTLVTTLGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKT
                     RAEVKFEGDTLVNRIELKGIGFKEDGNILGHKLEYNGTGFCFEAGADEDTHFRATSLET
                     LRNMVAAGSGITLLPALAVPPERKRDGVVYLPCIKPEPRRTIGLVYRPGSPLRSRYEQL
                     AEAIRARMDGHFDKVLKQAV"
     primer_bind     5343..5364
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gcggtggcac cggcagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
       61 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      121 ccacctacgg caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct
      181 ggcccaccct cgtgaccacc ctcggctacg gcctgaagtg cttcgcccgc taccccgacc
      241 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      301 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      361 acaccctggt gaaccgcatc gagctgaagg gcatcggctt caaggaggac ggcaacatcc
      421 tggggcacaa gctggagtac aacggcaccg gtttctgttt tgaagccggg gcggatgaag
      481 atacacactt ccgcgcgacc agcctggaaa ctctgcgcaa catggtggcg gcaggtagcg
      541 ggatcacttt actgccagcg ctggctgtgc cgccggagcg caaacgcgat ggggttgttt
      601 atctgccgtg cattaagccg gaaccacgcc gcactattgg tctggtttat cgtcctggct
      661 caccgctgcg cagccgctat gagcagctgg cagaggccat ccgcgcaaga atggatggcc
      721 atttcgataa agttttaaaa caggcggttt aaaagcttcg aattctgcag tcgacggtac
      781 cgcgggcccg ggatccaccg gatctagata actgatcata atcagccata ccacatttgt
      841 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
      901 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
      961 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     1021 caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     1081 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     1141 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     1201 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     1261 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     1321 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     1381 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     1441 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     1501 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1561 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1621 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     1681 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     1741 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     1801 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     1861 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     1921 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     1981 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     2041 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     2101 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     2161 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     2221 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     2281 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     2341 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     2401 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     2461 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     2521 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     2581 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     2641 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     2701 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     2761 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     2821 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     2881 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     2941 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     3001 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     3061 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     3121 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     3181 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     3241 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     3301 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     3361 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3421 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3481 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3541 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     3601 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3661 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3721 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3781 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3841 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3901 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3961 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4021 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     4081 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
     4141 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     4201 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     4261 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     4321 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     4381 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     4441 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     4501 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     4561 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     4621 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     4681 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc caccatggag
     4741 atggcaagcc agcagggcga gacgatgtcc ggaccgctgc acattggttt gattcccaca
     4801 gttggaccgt acctgctacc gcatattatc cctatgctgt accagacctt tccaaagctg
     4861 gaaatgtatc tgcatgaagc acagacccac cagttactgg cgcaactgga cagcggcaaa
     4921 ctcgattgcg tgatcctcgc actggtgaaa gagagcgaag cattcattga agtgccgttg
     4981 tttgatgagc caatgttgct ggctatctat gaagatcacc cgtgggcgaa ccgcgaatgc
     5041 gtaccgatgg ccgatctggc aggggaaaaa ctgctgatgc tggaagatgg tcactgtttg
     5101 cgcgatcagg caatgtccgc cggctacaac agcgacaacg tctatatcat ggccgacaag
     5161 cagaagaacg gcatcaaggc caacttcaag atccgccaca acgtcgagga cggcagcgtg
     5221 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     5281 gacaaccact acctgagctt ccagtccgtc ctgagcaaag accccaacga gaagcgcgat
     5341 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     5401 tacaacgtgg atggcggta
//
