LOCUS       sequence-431104-        5588 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fluorescent biosensor for the redox of mitochondrial specific 
            thioredoxin (Trx2).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5588)
  AUTHORS   Pang Y, Zhang H, Ai HW
  TITLE     Improved Red Fluorescent Redox Indicators for Monitoring Cytosolic 
            and Mitochondrial Thioredoxin Redox Dynamics.
  JOURNAL   Biochemistry. 2022 Mar 1;61(5):377-384. doi: 
            10.1021/acs.biochem.1c00634. Epub 2022 Feb 8.
  PUBMED    35133140
REFERENCE   2  (bases 1 to 5588)
  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: 
            "10.1021/acs.biochem.1c00634"; journalName: "Biochemistry"; date: 
            "2022-03-1- 1"; volume: "61"; issue: "5"; pages: "377-384"
COMMENT     Sequence Label: pMito-MtrxRFP2
FEATURES             Location/Qualifiers
     source          1..5588
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1084..1158
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1180..1251
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="SVLTPLLLRGLTGSARRLPVPRAK"
     CDS             1276..1293
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1297..1329
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(1333..1341,1342..1356)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     2511..2530
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2618..2752
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2642..2661)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2696..2715
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2865..2885)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2865..2883)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2904..2920)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2904..2920)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2917..2939)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2928..2944
                     /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(2952..2958,2959..2976,2977..2982))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2997..3018
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3135..3152)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3306..3894)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3386..3405)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4065..4856,4857..4925))
                     /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     4688..4707
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4926..5030)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5056..5511)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5193..5214)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5405..5424
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
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 accatgtccg tcctgacgcc gctgctgctg cggggcttga caggctcggc ccggcggctc
     1141 ccagtgccgc gcgccaagat ccattcgttg ggggatctgt ccgtcctgac gccgctgctg
     1201 ctgcggggct tgacaggctc ggcccggcgg ctcccagtgc cgcgcgccaa gatccattcg
     1261 ttgggggata ccggtcatca tcatcatcat catggtatgg ctagcatgac tggtggacag
     1321 caaatgggtc gggatctgta cgacgatgac gataaggatc cgagctcgag cacaaccttc
     1381 aatatccagg atggacctga ctttcaagac cgagtggtca acagtgagac accagtggtt
     1441 gtggatttcc acgcacagtg gtgtggaccc tgcaagatcc tggggccgag gttagagaag
     1501 atggtggcca agcagcacgg gaaggtggtg atggccaagg tggatattga tgaccacaca
     1561 gacctcgcca ttgagtatga ggtgtcagcg gtgcccactg tgctggccat gaagaatggg
     1621 gacgtggtgg acaagtttgt gggcatcaag gatgaggatc agttggaggc cttcctgaag
     1681 aagctgattg gcggtggatc cggtggcggc ggtagtggcg gtggagggag cggagggggg
     1741 ggaagcgggg gaggtggaag tggcggggga ggtagtgggg gcgagcttag ttgcgctata
     1801 agtcggctga gctcacccgt ggtttccgag cggatgtacc ccgaggacgg cgccctgaag
     1861 agcgagatca agaaggggct gaggctgaag gacggcggcc actacgccgc cgaggtcaag
     1921 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacatcgt cgacatcaag
     1981 ttggacatcg tgtcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
     2041 ggccgccact ccaccggcgg catggacgag ctgtacaagg gaggtacagg cgggagtctg
     2101 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
     2161 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     2221 ccctacgagg cctttcagac cgctaagctg aaggtgacca agggtggccc cctgcccttc
     2281 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacat taagcaccca
     2341 gccgacatcc ccgactactt caagctgtcc ttccccgagg gcttcaggtg ggagcgcgtg
     2401 atgaacttcg aggacggcgg tattattcac gttaaccagg actcctccct gcaggacggc
     2461 gtattcatct acaaggtgaa gctgcgcggc accaacttcc cccccgacgg ccccgtaatg
     2521 cagaagaaga ccatgggctg ggaggctacg cgtgaccaaa ggtgcaaaag tgtggcttaa
     2581 ctcgagcctc tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat
     2641 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     2701 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     2761 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc
     2821 ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta
     2881 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     2941 acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg tgcctaatga
     3001 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3061 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3121 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3181 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3241 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3301 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3361 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3421 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3481 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3541 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3601 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3661 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3721 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3781 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3841 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3901 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3961 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4021 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4081 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4141 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4201 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4261 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4321 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4381 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4441 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4501 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4561 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4621 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4681 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4741 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4801 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4861 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4921 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4981 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5041 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     5101 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     5161 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     5221 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     5281 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     5341 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     5401 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     5461 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
     5521 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     5581 agtcgacc
//
