LOCUS       Exported                6929 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses thiol redox-sensitive ratiometric sensor roGFP in the 
            mitochondrial matrix of mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    Matrix-roGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6929)
  AUTHORS   Waypa GB, Marks JD, Guzy R, Mungai PT, Schriewer J, Dokic D, 
            Schumacker PT
  TITLE     Hypoxia triggers subcellular compartmental redox signaling in 
            vascular smooth muscle cells.
  JOURNAL   Circ Res. 2010 Feb 19;106(3):526-35. doi: 
            10.1161/CIRCRESAHA.109.206334. Epub 2009 Dec 17.
  PUBMED    20019331
REFERENCE   2  (bases 1 to 6929)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1161/CIRCRESAHA.109.206334"; journalName: "Circ Res"; date: 
            "2010-02-19- 19"; volume: "106"; issue: "3"; pages: "526-35"
FEATURES             Location/Qualifiers
     source          1..6929
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        12..315
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        316..519
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     470..490
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             600..1319
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(645..666)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(906..925)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1253..1274
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1360..1494
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1384..1403)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1438..1457
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1637..1659)
                     /label=pGEX 3
                     /note="pGEX vectors, reverse primer"
     rep_origin      complement(4734..5322)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4814..4833)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5493..6353)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6116..6135
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6354..6458)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6526..6544
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     repeat_region   6572..6674
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     misc_signal     6762..6912
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"