LOCUS       sequence-330581-        6718 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Chemogenetic fluorescent H2O2 reporter targeted to the nucleus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6718)
  AUTHORS   Saeedi Saravi SS, Eroglu E, Waldeck-Weiermair M, Sorrentino A, 
            Steinhorn B, Belousov V, Michel T
  TITLE     Differential endothelial signaling responses elicited by 
            chemogenetic H2O2 synthesis.
  JOURNAL   Redox Biol. 2020 Sep;36:101605. doi: 10.1016/j.redox.2020.101605. 
            Epub 2020 Jun 16.
  PUBMED    32590330
REFERENCE   2  (bases 1 to 6718)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Redox 
            Biol."; date: "2020-09"; volume: "36:101605. doi"; pages: " 
            10.1016/j.redox.2020.101605. Epub 2020 Jun 16"
COMMENT     Sequence Label: HyPer7.2DAAO-NLS
FEATURES             Location/Qualifiers
     source          1..6718
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      100..555
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(187..206)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     397..418
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        581..685
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(686..754,755..1546)
                     /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     complement(904..923)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1717..2305
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2206..2225
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2459..2476
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2593..2614
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2629..2634,2635..2652,2653..2659)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2667..2683
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2672..2694
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2691..2707
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2691..2707
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2726..2746
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2728..2746
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    2859..2993
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2896..2915)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2950..2969
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3042..3062)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(3066..3086)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(3090..3110)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     4636..4655
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4895..4916
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5025..5046)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(5710..5728)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5711..5728)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(join(5763..6718,1..22))
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 atatacgcca tattgaattg gcggtcgact ggcgtaatag cgaagaggcc cgcaccgatc
       61 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
      121 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
      181 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
      241 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
      301 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
      361 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
      421 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
      481 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
      541 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
      601 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
      661 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
      721 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
      781 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
      841 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
      901 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
      961 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     1021 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     1081 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     1141 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     1201 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     1261 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     1321 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     1381 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     1441 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     1501 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     1561 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     1621 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     1681 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     1741 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     1801 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     1861 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     1921 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     1981 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2041 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2101 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2161 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2221 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     2281 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     2341 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     2401 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     2461 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     2521 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     2581 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     2641 gctcccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     2701 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctggg
     2761 taccgggccc aatgcattgg cgccgcggcc gcgaattaaa aaacctccca cacctccccc
     2821 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     2881 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2941 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctacgtaa
     3001 tacgactcac tatagttcta gaggctcgag gcaagctttt atacctttct cttctttttt
     3061 ggatctacct ttctcttctt ttttggatct acctttctct tcttttttgg atcgctctcc
     3121 ctagctgcgc cgtggtatct ttggaaggct tcatcgacca gctgagccac gtcctccacg
     3181 gctccccagg attgctgata ccctgcgcta ctaaagccgt aggcgtggac aagggtcact
     3241 tccttctcca ttgcagcccg tgcggaccct cttcccagag agagagggct cttggtccgg
     3301 tccaggggca agacgattct ttcagcctcc accctaggtc ctcctctcct ggcgggtctc
     3361 aggccaacgt tatgtctcag cacttcaatg ccctcgatag ttccatcgga gctaatggta
     3421 gggtcaagtc tcaggcagtg cttgagaatc ctttggactg tttcgggatt cacgctcagg
     3481 tcccaatcgc cgactccgta ggttccgcca cagatcactt ctcctcctgg tctaggaatg
     3541 atataagcgg gagaagcagg gtcggagcta tccattgtgc acctcttaca gggagatttg
     3601 acaagcacgg tctgtcccct aatgggctcg gcagcctggt catcgatgcc ggcaatggac
     3661 ttagctccca gcccggtggc gttgaccacg aggtcagctc catcgaaggc ttgctccagg
     3721 ctagtcactg ttctcctttc gaaggtagcc ccaagtttct gcagctctct ggcaaggtat
     3781 ctgcaatact tgggagcgtg gacgctcagg gtgtcgtatg tcactccaat ggctccaggt
     3841 ggacattcag aaggtggaag tggcctatag ttaggggtga tgtccttgta ccaatgtccc
     3901 aggaggccat cctcattctg ggcaaacctt ctggtccctt tcagccacat agcgtggcct
     3961 gtgggaacca gctccaccca tttcttgaag gtggactctt cccacttggc ttgtctgggt
     4021 ccgtctgtca gggtcataaa tggtgtccag ttagctccgg cccaggggct tgcgaaggtc
     4081 tgggagctga catcctcagg caagtctctg gccaggatat gcacggagta gccttttcta
     4141 gcgagaatca gggcgccaga gagcccgatg actccggagc ccagcacgac caccctcttc
     4201 tggctgtgac cactgctagc atcgcagatg aagctaacac catgcagttg gctttgcata
     4261 attgcggctt tcatggctga aagggcctta gggcggacaa agttgcggcg ataggccagg
     4321 acaacgcggc ggctaggcgg ggtgccttca aacgggataa tgctaaacag catgtggtcg
     4381 ttttcggtaa gggcagtggc aggcaagacg ctgattgcca agccactggc aaccatgtgg
     4441 cggatggtat tgatgcttga gccttgaagg gtgttggtga gaccctgaat gcgctgtttg
     4501 gcggcaagct cgctgcaact ggggccgttg tactccagct tgtgccccag gatgttgccg
     4561 tcctccttga agccgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac
     4621 ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg
     4681 acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag
     4741 cgggcgaagc acttcaggcc gtagctgagg gtggtcacga gggtgggcca gggcacgggc
     4801 agcttgccgg tggtgcagat cagcttcagg gtcagcttgc cgtaggtggc atcgccctcg
     4861 ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg
     4921 ggcaccaccc cggtgaacag ctcctcgccc ttgctgccgg tgccaccgct accgccatcc
     4981 acgttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg
     5041 accatgtgat cgcgcttctc gttggggtct ttgctcagga cggactggaa gctcaggtag
     5101 tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg
     5161 gcgagctgca cgctgccgtc ctcgacgttg tggcggatct tgaagttggc cttgatgccg
     5221 ttcttctgct tgtcggccat gatatagacg ttgttgctgt tgaagccggc ggagctaagc
     5281 acctggtcac gcatacagtt gccctcggtc agaagcagaa cttgttcttc gcccaacatg
     5341 cgagggctaa cggcatccaa ctcctcgaag ctatggccct tcggaacgat aacgaaaaac
     5401 ggctcgtcat acaaaggttc agtcacaata cccggctctt gaaacggctc tgccacaatg
     5461 atggcgtcaa cgtcgccgcg tttcagtgac tcggtaaggg tgtgggtgta attctcctcc
     5521 aacatcaaag gcaccttcgg ggcggtgcgg cgcaggctga cgatcaactt aggcagcaag
     5581 tacggggcga cggtaaagat caagcccacc ttaaaagcac cttccaattc gttttgctcc
     5641 tcattagcca ggtgcatggt ggcgaattcg aatcgatggg atcctgcaaa aagaacaagt
     5701 agcttgtatt ctatagtgtc acctaaatca agcttgctcc gacgtcccca ggcagaatgg
     5761 cggttcccta aacgagcatt gcttatatag acctcccatt aggcacgcct accgcccatt
     5821 tacgtcaatg gaacgcccat ttgcgtcatt gcccctcccc attgacgtca atggggatgt
     5881 acttggcagc catcgcgggc catttaccgc cattgacgtc aatgggagta ctgccaatgt
     5941 accctggcgt acttccaata gtaatgtact tgccaagtta ctattaatag atattgatgt
     6001 actgccaagt gggccattta ccgtcattga cgtcaatagg gggcgtgaga acggatatga
     6061 atgggcaatg agccatccca ttgacgtcaa tggtgggtgg tcctattgac gtcaatgggc
     6121 attgagccag gcgggccatt taccgtaatt gacgtcaatg ggggaggcgc catatacgtc
     6181 aataggaccg cccatatgac gtcaatagga aagaccatat atagagacca ttgacgtcaa
     6241 tgggggagtg gctatgggcg gtattaggaa gccccatata tggtatatgg gaccgcccat
     6301 tgggaggggc tatctacgtc aataggaaaa cccatatatg gaatactata tggcataggg
     6361 ccaatacata gtattgaacc tggccaatag ccatattggc atagggccat attggatatt
     6421 gcctatatat tgatcctggc atatagccaa tatggccgcc attattggca ccatgccaat
     6481 tcaatatggc ggacttggca cagggccaag acccctcctc aaacttggca ccgtgccaag
     6541 tagacccctc cccagttggc acagtgccag gtccgccata ttgaattggc atggtgccaa
     6601 gtccgccata ttgaattggc tggggtccag atccaccata ttgaattggc acgagtccat
     6661 atacgccata ttgaattggc acaggtccag atccaccata ttgaattggc atgagtcc
//
