LOCUS       sequence-431098-        5273 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 5273)
  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 5273)
  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: pBAD-MtrxRFP2
FEATURES             Location/Qualifiers
     source          1..5273
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             331..348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             352..384
                     /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(388..396,397..411)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     1566..1585
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1674..1691)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1844..1930
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2022..2049
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2068..2159
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2160..2228,2229..3020)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2378..2397)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3191..3779
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3680..3699
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3933..3950
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3965..4105
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4031..4050)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4191..4213
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4369..5247)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat ggggggttct catcatcatc atcatcatgg tatggctagc
      361 atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatccgagc
      421 tcgagcacaa cctttaatat ccaggatgga cctgactttc aagaccgagt ggtcaacagt
      481 gagacaccag tggttgtgga tttccacgca cagtggtgtg gaccctgcaa gatcctgggg
      541 ccgaggttag agaagatggt ggccaagcag cacgggaagg tggtgatggc caaggtggat
      601 attgatgacc acacagacct cgccattgag tatgaggtgt cagcggtgcc cactgtgctg
      661 gccatgaaga atggggacgt ggtggacaag tttgtgggca tcaaggatga ggatcagttg
      721 gaggccttcc tgaagaagct gattggcggt ggatccggtg gcggcggtag tggcggtgga
      781 gggagcggag gggggggaag cgggggaggt ggaagtggcg ggggaggtag tgggggcgag
      841 cttagttgcg ctataagtcg gctgagctca cccgtggttt ccgagcggat gtaccccgag
      901 gacggcgccc tgaagagcga gatcaagaag gggctgaggc tgaaggacgg cggccactac
      961 gccgccgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     1021 atcgtcgaca tcaagttgga catcgtgtcc cacaacgagg actacaccat cgtggaacag
     1081 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagggaggt
     1141 acaggcggga gtctggtgag caagggcgag gaggataaca tggccatcat caaggagttc
     1201 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     1261 gagggcgagg gccgccccta cgaggccttt cagaccgcta agctgaaggt gaccaagggt
     1321 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     1381 tacattaagc acccagccga catccccgac tacttcaagc tgtccttccc cgagggcttc
     1441 aggtgggagc gcgtgatgaa cttcgaggac ggcggtatta ttcacgttaa ccaggactcc
     1501 tccctgcagg acggcgtatt catctacaag gtgaagctgc gcggcaccaa cttccccccc
     1561 gacggccccg taatgcagaa gaagaccatg ggctgggagg ctacgcgtga ccaaaggtgc
     1621 aaaagtgtgg cttaaaagct tggctgtttt ggcggatgag agaagatttt cagcctgata
     1681 cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc
     1741 gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt
     1801 agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc
     1861 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     1921 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg
     1981 ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga
     2041 tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt
     2101 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     2161 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2221 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     2281 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2341 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2401 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2461 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2521 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     2581 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2641 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2701 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     2761 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     2821 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     2881 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     2941 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3001 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3061 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3121 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3181 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3241 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3301 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     3361 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3421 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3481 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3541 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3601 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3661 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3721 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3781 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3841 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3901 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3961 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
     4021 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     4081 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     4141 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4201 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagcaga tcaattcgcg
     4261 cgcgaaggcg aagcggcatg cataatgtgc ctgtcaaatg gacgaagcag ggattctgca
     4321 aaccctatgc tactccgtca agccgtcaat tgtctgattc gttaccaatt atgacaactt
     4381 gacggctaca tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc
     4441 ggtgcatttt ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc
     4501 gacggtggcg ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg
     4561 gtcctcgcgc cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg
     4621 cgacggcgac aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag
     4681 gtgatcgctg atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga
     4741 ctcgttaatc gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag
     4801 ctccgaatag cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa
     4861 atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca
     4921 gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc
     4981 gcgagcctcc ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc
     5041 acccggtcgg caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt
     5101 gagattgaga atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc
     5161 gttggcctca atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag
     5221 caggggatca ttttgcgctt cagccatact tttcatactc ccgccattca gag
//
