LOCUS       Exported                5531 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression of non-targeted ultrasensitive hydrogen 
            peroxide indicator HyPer7 for optical imaging.
ACCESSION   .
VERSION     .
KEYWORDS    pCS2+HyPer7
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5531)
  AUTHORS   Pak VV, Ezerina D, Lyublinskaya OG, Pedre B, Tyurin-Kuzmin PA, 
            Mishina NM, Thauvin M, Young D, Wahni K, Martinez Gache SA, 
            Demidovich AD, Ermakova YG, Maslova YD, Shokhina AG, Eroglu E, Bilan
            DS, Bogeski I, Michel T, Vriz S, Messens J, Belousov VV
  TITLE     Ultrasensitive Genetically Encoded Indicator for Hydrogen Peroxide 
            Identifies Roles for the Oxidant in Cell Migration and Mitochondrial
            Function.
  JOURNAL   Cell Metab. 2020 Mar 3;31(3):642-653.e6. doi: 
            10.1016/j.cmet.2020.02.003.
  PUBMED    32130885
REFERENCE   2  (bases 1 to 5531)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cmet.2020.02"; journalName: "Cell Metab"; date: 
            "2020-03-3- 3"; volume: "31"; issue: "3"; pages: "642-653.e6"
FEATURES             Location/Qualifiers
     source          1..5531
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     717..738
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(847..868)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1108..1127)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     promoter        complement(1561..1578)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1583..1717
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1607..1626)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1661..1680
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1830..1850)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1830..1848)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1869..1885)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1869..1885)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1882..1904)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1893..1909
                     /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(1917..1947)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1962..1983
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2100..2117)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2271..2859)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2351..2370)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3030..3890)
                     /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     3653..3672
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3891..3995)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4021..4476)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4158..4179)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4370..4389
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4554..5531
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"