LOCUS       sequence-189693-        6685 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  mammalian expression of a biosensor to monitor thioredoxin redox 
            dynamics, nuclear localization.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6685)
  AUTHORS   Fan Y, Makar M, Wang MX, Ai HW
  TITLE     Monitoring thioredoxin redox with a genetically encoded red 
            fluorescent biosensor.
  JOURNAL   Nat Chem Biol. 2017 Jun 26. doi: 10.1038/nchembio.2417.
  PUBMED    28671680
REFERENCE   2  (bases 1 to 6685)
  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"; journalName: "Nat Chem 
            Biol. 2017 Jun 26. doi: 10.1038/nchembio.2417."
COMMENT     Sequence Label: pNuc-TrxRFP1
FEATURES             Location/Qualifiers
     source          1..6685
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     729..748
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             799..819
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             823..843
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(883..901)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(884..901)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(921..938)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    927..1151
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1197..1625
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1284..1303)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1494..1515
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1634..1654)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1639..1968
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1819..1954
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1881..1900
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2035..2829
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2089..2108)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2699..2718
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3003..3124
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3040..3059)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3094..3113
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3173..3189)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3173..3189)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3186..3208)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3197..3213
                     /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(3221..3227,3228..3245,3246..3251))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3266..3287
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3404..3421)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3575..4163)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3655..3674)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4334..5125,5126..5194))
                     /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     4957..4976
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5195..5299)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5374..5393)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5565..5944
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5945..6148
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6099..6119
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6193..6212
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6193..6211
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      6228..6237
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6246..6263
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             6267..6299
                     /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(6303..6311,6312..6326)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
ORIGIN
        1 gagcttagtt gcgctataag tcggctgagc tcacccgtgg tttccgagcg gatgtacccc
       61 gaggacggcg ccctgaagag cgagatcaag aaggggctga ggctgaagga cggcggccac
      121 tacgccgccg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
      181 tacatcgtcg acatcaagtt ggacatcgtg tcccacaacg aggactacac catcgtggaa
      241 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaaggga
      301 ggtacaggcg ggagtctggt gagcaagggc gaggaggata acatggccat catcaaggag
      361 ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag
      421 ggcgagggcg agggccgccc ctacgaggcc tttcagaccg ctaagctgaa ggtgaccaag
      481 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag
      541 gcctacatta agcacccagc cgacatcccc gactacttca agctgtcctt ccccgagggc
      601 ttcaggtggg agcgcgtgat gaacttcgag gacggcggca ttattcacgt taaccaggac
      661 tcctccctgc aggacggcgt attcatctac aaggtgaagc tgcgcggcac caacttcccc
      721 cccgacggcc ccgtaatgca gaagaagacc atgggctggg aggctacgcg tgaccaaagg
      781 tgctccagtc aggctgatcc aaaaaagaag agaaaggtag acccgaagaa aaaacgaaag
      841 gtagatccaa agaagaagag aaagtaatct agagggccct attctatagt gtcacctaaa
      901 tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
      961 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     1021 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     1081 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     1141 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     1201 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     1261 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     1321 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     1381 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     1441 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     1501 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     1561 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     1621 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     1681 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     1741 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     1801 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     1861 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     1921 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     1981 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     2041 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     2101 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     2161 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     2221 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     2281 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     2341 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     2401 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     2461 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     2521 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     2581 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     2641 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     2701 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     2761 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     2821 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     2881 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     2941 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     3001 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3061 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3121 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     3181 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     3241 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     3301 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3361 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3421 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3481 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3541 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3601 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3661 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3721 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3781 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3841 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3901 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3961 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4021 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4081 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4141 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4201 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4261 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4321 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4381 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4441 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4501 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4561 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4621 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4681 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4741 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4801 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4861 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4921 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4981 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5041 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5101 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5161 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5221 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5281 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     5341 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     5401 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     5461 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
     5521 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
     5581 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     5641 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     5701 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     5761 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     5821 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     5881 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     5941 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     6001 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     6061 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     6121 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
     6181 gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccgcc accatgggag
     6241 gttctcatca tcatcatcat catggtatgg ctagcatgac tggtggacag caaatgggtc
     6301 gggatctgta cgacgatgac gataaggatc cgagctcgag catggttaaa cagatcgaat
     6361 ctaaaaccgc gttccaggaa gcgctggacg cggcgggtga caaactggtt gttgttgact
     6421 tctctgcgac ctggtgcggt ccgtgcaaaa tgatcaaacc gttcttccac tctctgtctg
     6481 aaaaatactc taacgttatc ttcctggaag ttgacgttga cgactcccag gacgttgcgt
     6541 ctgaatccga agttaaatcc atgccgacct tccagttctt caaaaaaggt cagaaagttg
     6601 gtgaattctc tggtgcgaac aaagaaaaac tggaagcgac catcaacgaa ctggttggtg
     6661 gcagtggtgg cggcggtagt ggcgg
//
