LOCUS       Exported                6673 bp ds-DNA     circular SYN 30-APR-2021
DEFINITION  Chemogenetic fluorescent H2O2 reporter targeted to the cytoplasm.
ACCESSION   .
VERSION     .
KEYWORDS    HyPer7.2DAAO-NES
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6673)
  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 6673)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Apr 30, 2021 from SnapGene Viewer 5.1.4.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Redox
            Biol."; date: "2020-09"; pages: "
            10.1016/j.redox.2020.101605. Epub 2020 Jun 16"
FEATURES             Location/Qualifiers
     source          1..6673
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian 
                     cytomegalovirusmajor immediate early transcription unit 
                     IE94"
                     /note="color: #ffffff"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /note="color: #ffffff"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
                     /note="color: #a020f0; direction: RIGHT"
     misc_feature    1084..2520
                     /label=HyPer7.2
                     /note="color: #ffcc00"
     misc_signal     1132..1134
                     /label=L17V
                     /note="color: #a6acb3"
     misc_feature    1207..1209
                     /label=M42V
                     /note="color: #a6acb3"
     primer_bind     1695..1716
                     /label=EGFP-C
                     /note="EGFP, forward primer"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     complement(1825..1846)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(2086..2105)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
                     /note="color: #a020f0; direction: LEFT"
     misc_feature    2527..3627
                     /label=DAAO
                     /note="color: #a6acb3"
     misc_feature    3628..3654
                     /label=NES
                     /note="color: #993366; direction: RIGHT"
     promoter        complement(3681..3698)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     polyA_signal    3703..3837
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(3727..3746)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     3781..3800
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     complement(3950..3970)
                     /label=T3
                     /note="T3 promoter, forward primer"
                     /note="color: #a020f0; direction: LEFT"
     promoter        complement(3950..3968)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(3989..4005)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(3989..4005)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     complement(4002..4024)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    4013..4029
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator 
                     toinhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /note="color: #31849b"
     promoter        complement(4037..4067)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="color: #ffffff; direction: LEFT"
     protein_bind    4082..4103
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the 
                     presenceof cAMP."
                     /note="color: #31849b"
     primer_bind     complement(4220..4237)
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: #a020f0; direction: LEFT"
     rep_origin      complement(4391..4979)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     complement(4471..4490)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: #a020f0; direction: LEFT"
     CDS             complement(5150..6010)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="color: #993366"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5773..5792
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: #a020f0; direction: RIGHT"
     promoter        complement(6011..6115)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      complement(6141..6596)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; 
                     arrowindicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     primer_bind     complement(6278..6299)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     6490..6509
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
                     /note="color: #a020f0; direction: RIGHT"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat cccatcgatt cgaattcgcc
     1081 accatgcacc tggctaatga ggagcaaaac gaattggaag gtgcttttaa ggtgggcttg
     1141 atctttaccg tcgccccgta cttgctgcct aagttgatcg tcagcctgcg ccgcaccgcc
     1201 ccgaaggtgc ctttgatgtt ggaggagaat tacacccaca cccttaccga gtcactgaaa
     1261 cgcggcgacg ttgacgccat cattgtggca gagccgtttc aagagccggg tattgtgact
     1321 gaacctttgt atgacgagcc gtttttcgtt atcgttccga agggccatag cttcgaggag
     1381 ttggatgccg ttagccctcg catgttgggc gaagaacaag ttctgcttct gaccgagggc
     1441 aactgtatgc gtgaccaggt gcttagctcc gccggcttca acagcaacaa cgtctatatc
     1501 atggccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacgtcgag
     1561 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1621 gtgctgctgc ccgacaacca ctacctgagc ttccagtccg tcctgagcaa agaccccaac
     1681 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1741 atggacgagc tgtacaacgt ggatggcggt agcggtggca ccggcagcaa gggcgaggag
     1801 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1861 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctg
     1921 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctcagctac
     1981 ggcctgaagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     2041 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     2101 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     2161 ggcatcggct tcaaggagga cggcaacatc ctggggcaca agctggagta caacggcccc
     2221 agttgcagcg agcttgccgc caaacagcgc attcagggtc tcaccaacac ccttcaaggc
     2281 tcaagcatca ataccatccg ccacatggtt gccagtggct tggcaatcag cgtcttgcct
     2341 gccactgccc ttaccgaaaa cgaccacatg ctgtttagca ttatcccgtt tgaaggcacc
     2401 ccgcctagcc gccgcgttgt cctggcctat cgccgcaact ttgtccgccc taaggccctt
     2461 tcagccatga aagccgcaat tatgcaaagc caactgcatg gtgttagctt catctgcgat
     2521 gctagcagtg gtcacagcca gaagagggtg gtcgtgctgg gctccggagt catcgggctc
     2581 tctggcgccc tgattctcgc tagaaaaggc tactccgtgc atatcctggc cagagacttg
     2641 cctgaggatg tcagctccca gaccttcgca agcccctggg ccggagctaa ctggacacca
     2701 tttatgaccc tgacagacgg acccagacaa gccaagtggg aagagtccac cttcaagaaa
     2761 tgggtggagc tggttcccac aggccacgct atgtggctga aagggaccag aaggtttgcc
     2821 cagaatgagg atggcctcct gggacattgg tacaaggaca tcacccctaa ctataggcca
     2881 cttccacctt ctgaatgtcc acctggagcc attggagtga catacgacac cctgagcgtc
     2941 cacgctccca agtattgcag ataccttgcc agagagctgc agaaacttgg ggctaccttc
     3001 gaaaggagaa cagtgactag cctggagcaa gccttcgatg gagctgacct cgtggtcaac
     3061 gccaccgggc tgggagctaa gtccattgcc ggcatcgatg accaggctgc cgagcccatt
     3121 aggggacaga ccgtgcttgt caaatctccc tgtaagaggt gcacaatgga tagctccgac
     3181 cctgcttctc ccgcttatat cattcctaga ccaggaggag aagtgatctg tggcggaacc
     3241 tacggagtcg gcgattggga cctgagcgtg aatcccgaaa cagtccaaag gattctcaag
     3301 cactgcctga gacttgaccc taccattagc tccgatggaa ctatcgaggg cattgaagtg
     3361 ctgagacata acgttggcct gagacccgcc aggagaggag gacctagggt ggaggctgaa
     3421 agaatcgtct tgcccctgga ccggaccaag agccctctct ctctgggaag agggtccgca
     3481 cgggctgcaa tggagaagga agtgaccctt gtccacgcct acggctttag tagcgcaggg
     3541 tatcagcaat cctggggagc cgtggaggac gtggctcagc tggtcgatga agccttccaa
     3601 agataccacg gcgcagctag ggagagcctg ccccccctgg agcggctgac cctgtaaaag
     3661 cttgcctcga gcctctagaa ctatagtgag tcgtattacg tagatccaga catgataaga
     3721 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3781 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3841 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     3901 ttcgcggccg cggcgccaat gcattgggcc cggtacccag cttttgttcc ctttagtgag
     3961 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4021 cgctcacaat tccacacaac atacgagccg ggagcataaa gtgtaaagcc tggggtgcct
     4081 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4141 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4201 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4261 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4321 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4381 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4441 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4501 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4561 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4621 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4681 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4741 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4801 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4861 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4921 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4981 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5041 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5101 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5161 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5221 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5281 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5341 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5401 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5461 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5521 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5581 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5641 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5701 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5761 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5821 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5881 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5941 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6001 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6061 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6121 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     6181 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     6241 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     6301 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     6361 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     6421 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     6481 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     6541 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca
     6601 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     6661 acgccagtcg acc
//
