LOCUS       sequence-310656-        7760 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Peroxynitrite sensor pnGFP-Ultra in mammalian cells (use with 
            pMAH-POLY-eRF1(E55D)).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7760)
  AUTHORS   Chen Z, Zhang S, Li X, Ai HW
  TITLE     A high-performance genetically encoded fluorescent biosensor for 
            imaging physiological peroxynitrite.
  JOURNAL   Cell Chem Biol. 2021 Jan 27. pii: S2451-9456(21)00013-1. doi: 
            10.1016/j.chembiol.2021.01.013.
  PUBMED    33581056
REFERENCE   2  (bases 1 to 7760)
  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: "Cell Chem 
            Biol. 2021 Jan 27. pii: S2451-9456(21)00013-1. doi: 
            10.1016/j.chembiol.2021.01.013."
COMMENT     Sequence Label: pMAH-pnGFP-Ultra
FEATURES             Location/Qualifiers
     source          1..7760
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(111..201)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     promoter        complement(313..561)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        complement(850..940)
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     promoter        complement(1052..1300)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     enhancer        1559..1862
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1863..2066
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2017..2037
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2063..2087
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        2297..2315
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2297..2314
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     2328..2347
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      2383..2392
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2625..2646
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2758..2779)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             3151..3168
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3200..3217)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3206..3430
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3476..3904
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3563..3582)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3773..3794
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3913..3933)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3918..4247
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4098..4233
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4160..4179
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4296..5321
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4296..5321
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    5451..5572
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5488..5507)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5542..5561
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5621..5637)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5621..5637)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5634..5656)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5645..5661
                     /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)."
     promoter        complement(join(5669..5675,5676..5693,5694..5699))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5714..5735
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5852..5869)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6023..6611)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6103..6122)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6782..7573,7574..7642))
                     /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     7405..7424
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7643..7747)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctcgactcgc tagtgttcca aaaaatggtg ggggacggat tcgaaccgcc gaacccaaag
       61 ggagcggatt tagagtccgc ggcgtttggc cacttcgcta ccccaccaga gtggtctcat
      121 acagaactta taagattccc aaatccaaag acatttcacg tttatggtga tttcccagaa
      181 cacatagcga catgcaaata tgaattgatg actgcctgat ggagaaaaaa tggtggggga
      241 cggattcgaa ccgccgaacc caaagggagc ggatttagag tccgcggcgt ttggccactt
      301 cgctacccca ccggtgtttc gtcctttcca caagatatat aaagccaaga aatcgaaata
      361 ctttcaagtt acggtaagca tatgatagtc cattttaaaa cataatttta aaactgcaaa
      421 ctacccaaga aattattact ttctacgtca cgtattttgt actaatatct ttgtgtttac
      481 agtcaaatta attctaatta tctctctaac agccttgtat cgtatatgca aatatgaagg
      541 aatcatggga aataggccct cttcctgccc gaccttcgcg atgtacgggc cagatatacg
      601 ccaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
      661 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      721 aatatgaccg ccatgttggc tcgactcgct agtgttccaa aaaatggtgg gggacggatt
      781 cgaaccgccg aacccaaagg gagcggattt agagtccgcg gcgtttggcc acttcgctac
      841 cccaccagag tggtctcata cagaacttat aagattccca aatccaaaga catttcacgt
      901 ttatggtgat ttcccagaac acatagcgac atgcaaatat gaattgatga ctgcctgatg
      961 gagaaaaaat ggtgggggac ggattcgaac cgccgaaccc aaagggagcg gatttagagt
     1021 ccgcggcgtt tggccacttc gctaccccac cggtgtttcg tcctttccac aagatatata
     1081 aagccaagaa atcgaaatac tttcaagtta cggtaagcat atgatagtcc attttaaaac
     1141 ataattttaa aactgcaaac tacccaagaa attattactt tctacgtcac gtattttgta
     1201 ctaatatctt tgtgtttaca gtcaaattaa ttctaattat ctctctaaca gccttgtatc
     1261 gtatatgcaa atatgaagga atcatgggaa ataggccctc ttcctgcccg accttcgcga
     1321 tgtacgggcc agatatacgc caatattggc cattagccat attattcatt ggttatatag
     1381 cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca
     1441 tttatattgg ctcatgtcca atatgaccgc catgttggct cgagcattga ttattgacta
     1501 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     1561 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     1621 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     1681 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     1741 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     1801 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     1861 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
     1921 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     1981 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
     2041 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc
     2101 atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc cgcggccggg
     2161 aacggtgcat tggaacgcgg attccccgtg ccaagagtga cgtaagtacc gcctatagag
     2221 tctataggcc cacccccttg gcttcgttag aacgcggcta caattaatac ataaccttat
     2281 gtatcataca catacgattt aggtgacact atagaataac atccactttg cctttctctc
     2341 cacaggtgtc cactcccagg tccaactgca cggaagcttg ccgccaccat gggctccagc
     2401 aagaagtaca acagccacaa ggtctatatc accgccgaca agcagaagaa cggcatcaag
     2461 gcgaacttcc agatccgcca caacgtggag gacggcagcg tgcagctcgc cgaccactac
     2521 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     2581 acccaggccg tgctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     2641 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgcacaaggt ggacggcgga
     2701 tccggtggca ccggagtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2761 gtcgagctgg acggcgacgt aaacggccac aagttccgcg tgaggggcga aggcgagggc
     2821 gatgccacca acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     2881 ccctggccca ccctcgtgac caccttgacc tagggcgtgc agtgcttctc ccgctacccc
     2941 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3001 cgcaccatct tcttcaagga cgacggcacc tacaagaccc gcgccgaggt gaagttcgag
     3061 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3121 atcctggggc acaagctgga gtacaactcg caccatcacc atcaccatta ataagggccc
     3181 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3241 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3301 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3361 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3421 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3481 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3541 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3601 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3661 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3721 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3781 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3841 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3901 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3961 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4021 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4081 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4141 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4201 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4261 cttgtatatc cattttcgga tctgatcagc acgtgatgaa aaagcctgaa ctcaccgcga
     4321 cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct
     4381 cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc
     4441 gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat
     4501 cggccgcgct cccgattccg gaagtgcttg acattgggga attcagcgag agcctgacct
     4561 attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc
     4621 ccgctgttct gcagccggtc gcggaggcca tggatgcgat cgctgcggcc gatcttagcc
     4681 agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg
     4741 atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca
     4801 ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc
     4861 ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg
     4921 gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg
     4981 tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact
     5041 tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat atgctccgca
     5101 ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg
     5161 cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa
     5221 tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc gccgatagtg
     5281 gaaaccgacg ccccagcact cgtccgaggg caaaggaata gcacgtgcta cgagatttcg
     5341 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5401 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5461 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5521 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5581 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5641 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5701 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5761 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5821 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5881 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5941 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6001 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6061 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6121 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6181 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6241 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6301 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6361 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6421 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6481 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6541 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6601 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6661 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6721 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6781 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6841 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6901 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6961 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7021 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7081 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7141 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7201 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7261 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7321 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7381 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7441 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7501 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7561 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7621 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7681 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7741 ttccgcgcac atttccccga
//
