LOCUS       sequence-227465-        7550 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mitochondrial Grx1-roGFP2 (retroviral 
            vector).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7550)
  AUTHORS   Gutscher M, Pauleau AL, Marty L, Brach T, Wabnitz GH, Samstag Y, 
            Meyer AJ, Dick TP
  TITLE     Real-time imaging of the intracellular glutathione redox potential.
  JOURNAL   Nat Methods. 2008 Jun;5(6):553-9. doi: 10.1038/nmeth.1212. Epub 2008
            May 11.
  PUBMED    18469822
REFERENCE   2  (bases 1 to 7550)
  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"; doi: 
            "10.1038/nmeth.1212"; journalName: "Nat Methods"; date: "2008-06"; 
            volume: "5"; issue: "6"; pages: "553-9"
COMMENT     Sequence Label: pLPCX mito Grx1-roGFP2
FEATURES             Location/Qualifiers
     source          1..7550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(1189..1191,1192..1905)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1231..1252)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1492..1511)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1839..1860
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             1951..2544
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2647..2669)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    2797..2830
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2890..2923
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2943..2962
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3043..3060)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3214..3802)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3294..3313)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3973..4764,4765..4833))
                     /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     4596..4615
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4834..4938)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5006..5024
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     LTR             5626..5801
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    5864..6063
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             6264..6680
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6610..6632
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6648..6664
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             6777..7376
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6777..6796)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7273..7293
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagat ggcctccact cgtgtcctcg cctctcgcct ggcctcccag
      601 atggctgctt ccgccaaggt tgcccgccct gctgtccgcg ttgctcaggt cagcaagcgc
      661 accatccaga ctggctcccc cctccagacc ctcaagcgca cccagatgac ctccatcgtc
      721 aacgccacca cccgccaggc tttccagaag cgcgcctact cttccggatc cgctcaagag
      781 tttgtgaact gcaaaatcca gcctgggaag gtggttgtgt tcatcaagcc cacctgcccg
      841 tactgcagga gggcccaaga gatcctcagt caattgccca tcaaacaagg gcttctggaa
      901 tttgtcgata tcacagccac caaccacact aacgagattc aagattattt gcaacagctc
      961 acgggagcaa gaacggtgcc tcgagtcttt attggtaaag attgtatagg cggatgcagt
     1021 gatctagtct ctttgcaaca gagtggggaa ctgctgacgc ggctaaagca gattggagct
     1081 ctgcagacta gtggtggttc aggtggtggt ggttcaggtg gtggtggttc aggtggagga
     1141 ggatcaggag gaggaggatc aggaggagga ggatcaggag gagaattcgt gagcaagggc
     1201 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1261 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     1321 aagttcatct ccaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     1381 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     1441 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1501 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1561 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1621 tacaactgcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     1681 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     1741 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacctgc
     1801 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1861 accgccgccg ggatcactct cggcatggac gagctgtaca agtaaatcga taaaataaaa
     1921 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
     1981 gctagcttaa gtaacgccat tttgcaaggc atggaaaaat acataactga gaatagagaa
     2041 gttcagatca aggtcaggaa cagatggaac agctgaatat gggccaaaca ggatatctgt
     2101 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggaacagctg aatatgggcc
     2161 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc
     2221 ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc cagggtgccc
     2281 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
     2341 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcgggg
     2401 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc
     2461 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
     2521 acccgtcagc gggggtcttt catttggggg ctcgtccggg atcgggagac ccctgcccag
     2581 ggaccaccga cccaccaccg ggaggtaagc tggctgcctc gcgcgtttcg gtgatgacgg
     2641 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     2701 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
     2761 catgacccag tcacgtagcg atagcggagt gtactgataa cttcgtataa tgtatgctat
     2821 acgaagttat taggtctgaa gaggagttta cgtccagcca agctagggcc gcgatccgga
     2881 acccttaata taacttcgta taatgtatgc tatacgaagt tatcagtact ggcttaacta
     2941 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     3001 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct
     3061 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3121 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3181 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3241 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3301 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3361 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3421 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3481 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3541 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3601 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     3661 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3721 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3781 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3841 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3901 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3961 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4021 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4081 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4141 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4201 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4261 tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg
     4321 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4381 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4441 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4501 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4561 gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac
     4621 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4681 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4741 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4801 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4861 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4921 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     4981 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga
     5041 attgctagca attgctagca attgctagca attcatacca gatcaccgaa aactgtcctc
     5101 caaatgtgtc cccctcacac tcccaaattc gcgggcttct gcctcttaga ccactctacc
     5161 ctattcccca cactcaccgg agccaaagcc gcggcccttc cgtttctttg cttttgaaag
     5221 accccacccg taggtggcaa gctagcttaa gtaacgccac tttgcaaggc atggaaaaat
     5281 acataactga gaatagaaaa gttcagatca aggtcaggaa caaagaaaca gctgaatacc
     5341 aaacaggata tctgtggtaa gcggttcctg ccccggctca gggccaagaa cagatgagac
     5401 agctgagtga tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcggggc
     5461 caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagtg aatcatcaga
     5521 tgtttccagg gtgccccaag gacctgaaaa tgaccctgta ccttatttga actaaccaat
     5581 cagttcgctt ctcgcttctg ttcgcgcgct tccgctctcc gagctcaata aaagagccca
     5641 caacccctca ctcggcgcgc cagtcttccg atagactgcg tcgcccgggt acccgtattc
     5701 ccaataaagc ctcttgctgt ttgcatccga atcgtggtct cgctgttcct tgggagggtc
     5761 tcctctgagt gattgactac ccacgacggg ggtctttcat ttgggggctc gtccgggatt
     5821 tggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg ccagcaactt
     5881 atctgtgtct gtccgattgt ctagtgtcta tgtttgatgt tatgcgcctg cgtctgtact
     5941 agttagctaa ctagctctgt atctggcgga cccgtggtgg aactgacgag ttctgaacac
     6001 ccggccgcaa ccctgggaga cgtcccaggg actttggggg ccgtttttgt ggcccgacct
     6061 gaggaaggga gtcgatgtgg aatccgaccc cgtcaggata tgtggttctg gtaggagacg
     6121 agaacctaaa acagttcccg cctccgtctg aatttttgct ttcggtttgg aaccgaagcc
     6181 gcgcgtcttg tctgctgcag cgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt
     6241 ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag
     6301 gtcactggaa agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac
     6361 gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg
     6421 gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc
     6481 atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc
     6541 ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc
     6601 cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc
     6661 tcactccttc tctaggcgcc ggaatgatcc cccgggctgc aggtcggccg ccacgaccgg
     6721 tgccgccacc atcccctgac ccacgcccct gacccctcac aaggagacga ccttccatga
     6781 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca
     6841 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc
     6901 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     6961 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     7021 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     7081 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     7141 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     7201 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     7261 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     7321 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacgcc
     7381 cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatggc tccgaccgaa
     7441 gccacccggg gcggccccgc cgaccccgca cccgcccccg aggcccaccg actctagtaa
     7501 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
//
