LOCUS       sequence-37023-e        8075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8075)
  AUTHORS   Hung YP, Albeck JG, Tantama M, Yellen G
  TITLE     Imaging Cytosolic NADH-NAD(+) Redox State with a Genetically Encoded
            Fluorescent Biosensor.
  JOURNAL   Cell Metab. 2011 Oct 5;14(4):545-54.
  PUBMED    21982714
REFERENCE   2  (bases 1 to 8075)
  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: "Cell 
            Metab."; date: "2011-10-5"; volume: "14(4)"; pages: "545-54"
COMMENT     Sequence Label: GW1-Peredox-mCherry-NLS
FEATURES             Location/Qualifiers
     source          1..8075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(133..744,745..765,766..1500,1501..1503,1504..2133,
                     2134..2178)
                     /codon_start=1
                     /product="genetically encoded fluorescent biosensor of the 
                     cytosolic NADH-NAD+ redox state (Hung et al., 2011)"
                     /label=Peredox
                     /note="consists of two pH-insensitive mutant copies of the 
                     Thermus aquaticus redox-sensing repressor Rex (T-Rex) 
                     separated by circularly permuted monomeric T-Sapphire 
                     (cpmTS), a pH-resistant GFP variant"
                     /translation="KVPEAAISRLITYLRILEELEAQGVHRTASEQLGELAQVTAFQVD
                     KDLSYFGSYGTDGVGYTVPVLKRELRHILGLNRKWGLCIVGMGRLGSALADWPGFGESF
                     ELRGFFDVDPGMVGRPVRGGVIEHVDLLPQRVPGRIEIALLTVPREAAQKAADLLVAAG
                     IKGILNFAPVVLEVPKEVAVENVDILAGLTRLSFAILNPTWSAAGGHGFTAHNVYIMAD
                     KQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSIQSKLSKDPNEK
                     RDHMVLLEFVTAAGITHGMDELYKGGTGGSMVSKGEELFTGVVPILVELDGDVNGHKFS
                     VSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFSYGVMVFARYPDHMKQHDFFKSA
                     MPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNTK
                     VPEAAISRLITYLRILEELEAQGVHRTASEQLGELAQVTAFQVDEDLSYFGSYGTDGVG
                     YTVPVLKRELRHILGLNRKWGLCIVGMGRLGSALADWPGFGESFELRGFFDVDPGMVGR
                     PVRGGVIEHVDLLPQRVPGRIEIALLTVPREAAQKAADLLVAAGIKGILNFAPVVLEVP
                     KEVAVENVDFLAGLTRLSFAILNPKWREEMMGSGTGGNASDGGGSGG"
     CDS             2179..2886
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2324..2342)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2592..2611
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2887..2907
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3041..3175
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3078..3097)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3132..3151
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3199..3215)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3199..3215)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3212..3234)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3223..3239
                     /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(3247..3253,3254..3271,3272..3277))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3292..3313
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3430..3447)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3601..4189)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3681..3700)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4360..5151,5152..5220))
                     /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     4983..5002
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     5430..5448
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5486..5508)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5608..5627
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5859..5881
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5873..5890
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5874..5890
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        6439..6818
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6819..7022
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6973..6993
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7019..7043
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          7158..7984
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
ORIGIN
        1 tccgagtccg ataccgataa agattctgaa tcggagtgag tgccgcgtcc gagagcgttc
       61 cctgtccacg tccaccatcg gcggagcagg tgtgcctgag gcctctagat tgggcccacc
      121 atggttgcta gcaaagttcc tgaagcagcc atttccagac tgattactta cctgagaatt
      181 ctggaagagc tggaagccca aggtgtacac cgcaccgcct ccgagcaact gggcgagctg
      241 gcccaggtca ccgccttcca ggttgataag gacctgtcct acttcggctc ctacggcact
      301 gacggcgttg gctacactgt accggtcctc aagagagaac tgcgccatat cctcggtctg
      361 aaccgcaaat ggggcctgtg catcgtgggg atgggccgcc tgggatccgc tcttgctgat
      421 tggcctggtt ttggcgagag cttcgagctg cgcggtttct tcgatgttga cccaggtatg
      481 gttggtcggc cggttcgcgg tggtgttatc gaacacgttg atctgctgcc ccagcgcgta
      541 cctggtcgca tcgaaatcgc tctgctgacg gttccacgcg aggcagcaca gaaggctgcc
      601 gacctgctgg ttgccgctgg catcaagggc atcctgaact tcgctccagt tgtactggag
      661 gttcccaagg aagtggctgt tgagaacgtt gacatcctgg ccggtctgac gcgtctgagc
      721 tttgccattc tgaaccccac gtggagcgca gcaggtgggc atggtttcac tgctcacaac
      781 gtgtatatca tggctgacaa acagaagaac ggtatcaagg ccaacttcaa gatccgccac
      841 aacatcgaag acggcggcgt tcagctggct gatcactacc agcagaacac cccaatcggc
      901 gacggccctg ttctgctgcc tgacaaccac tacttaagca tccagtctaa actgtccaaa
      961 gacccaaacg agaagcgcga ccacatggtt ctgctggaat ttgttaccgc cgctggcatc
     1021 acccacggca tggacgaact gtacaagggc ggcaccgggg gatctatggt ttccaaaggc
     1081 gaggaactgt tcactggtgt tgttcctatc ctggttgaac tggacggcga cgttaacggc
     1141 cacaaattca gcgttagcgg ggaaggcgaa ggagacgcta cctacggcaa gctgactctg
     1201 aagttcatct gcaccactgg caagctgcct gttccttggc ctaccctggt tactaccttc
     1261 tcctatggtg ttatggtttt cgccagatat cctgaccaca tgaagcagca cgacttcttc
     1321 aagagcgcca tgcctgaggg ctacgttcag gaacgcacca tctttttcaa ggacgacggc
     1381 aactataaga cacgcgctga ggttaagttc gagggcgaca ctctggttaa ccgcatcgag
     1441 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg gccataagct tgaatataac
     1501 accaaggtac ctgaagccgc catcagccgc ctcatcactt acctccgcat cctggaggag
     1561 ctcgaagccc aaggcgttca ccgcactgca agcgagcagc tgggcgaact ggcacaggtt
     1621 acagcattcc aggttgacga agatctgtcc tatttcggtt cctacggcac cgacggtgtt
     1681 ggctacacag tccctgtcct gaagcgcgag ctgcgccaca tcctgggcct gaaccgcaag
     1741 tggggcttat gcatcgttgg catgggccgc ctcggtagtg cactggcaga ctggccgggc
     1801 tttggtgaat ctttcgaact gcgcggcttc ttcgacgttg atccaggcat ggttgggcgc
     1861 cctgttcgcg gtggcgttat cgagcatgtt gacctcctgc cacagcgcgt cccgggtcgc
     1921 atcgagatcg ccctgctgac cgttcctcgc gaagctgcac agaaggcagc cgacttacta
     1981 gtcgccgctg gtatcaaggg aattctgaac ttcgcccctg ttgttctgga ggttcctaag
     2041 gaagttgctg ttgaaaacgt tgatttcctg gccggcctga cccgcctgag cttcgcaatc
     2101 ctgaatccta agtggcgcga agaaatgatg ggttccggaa ctgggggcaa cgcctctgac
     2161 ggtggtgggt ctggtggtat ggtgagcaag ggcgaggagg ataacatggc catcatcaag
     2221 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
     2281 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     2341 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
     2401 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
     2461 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
     2521 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
     2581 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
     2641 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
     2701 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
     2761 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
     2821 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgaactg
     2881 tacaagccta aaaagaaacg taaagttgaa gatgcttgag tcgacgaatt cattgatcat
     2941 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3001 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3061 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3121 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgtac
     3181 cgagctcgcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     3241 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3301 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3361 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3421 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3481 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3541 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3601 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3661 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3721 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3781 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3841 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3901 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3961 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4021 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     4081 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4141 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4201 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4261 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4321 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4381 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4441 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4501 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4561 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4621 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4681 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4741 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4801 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4861 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4921 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4981 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5041 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5101 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5161 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5221 actcttcctt tttcaataag cggccgcggc catgccggcc actagtctcg agttattatt
     5281 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5341 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     5401 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     5461 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     5521 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     5581 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     5641 atatgaagac gtcgcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg
     5701 cctctgcata aataaaaaaa attagtcagc gcgccattcg ccattcaggc tgcgcaactg
     5761 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     5821 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     5881 gacggccagt gccatctgca gtgaataata aaatgtgtgt ttgtccgaaa tacgcgtttg
     5941 agatttctgt cccgactaaa ttcatgtcgc gcgatagtgg tgtttatcgc cgatagagat
     6001 ggcgatattg gaaaaatcga tatttgaaaa tatggcatat tgaaaatgtc gccgatgtga
     6061 gtttctgtgt aactgatatc gccatttttc caaaagttga tttttgggca tacgcgatat
     6121 ctggcgatac gcttatatcg tttacggggg atggcgatag acgcctttgg tgacttgggc
     6181 gattctgtgt gtcgcaaata tcgcagtttc gatataggtg acagacgata tgaggctata
     6241 tcgccgatag aggcgacatc aagctggcac atggccaatg catatcgatc tatacattga
     6301 atcaatattg gccattagcc atattattca ttggttatat agcataaatc aatattggct
     6361 attggccatt gcatacgttg tatccatatc ataatatgta catttatatt ggctcatgtc
     6421 caacattacc gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     6481 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     6541 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     6601 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     6661 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     6721 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     6781 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     6841 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     6901 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     6961 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     7021 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     7081 gaagacaccg ggaccgatcc agcctccgcg gccgggaacg gtgcattgga acgcggattc
     7141 cccgtgccaa gagtgacgta agtaccgcct atagagtcta taggcccacc cccttggctt
     7201 cttatgcatg ctatactgtt tttggcttgg ggtctataca cccccgcttc ctcatgttat
     7261 aggtgatggt atagcttagc ctataggtgt gggttattga ccattattga ccactcccct
     7321 attggtgacg atactttcca ttactaatcc ataacatggc tctttgccac aactctcttt
     7381 attggctata tgccaataca ctgtccttca gagactgaca cggactctgt atttttacag
     7441 gatggggtct catttattat ttacaaattc acatatacaa caccaccgtc cccagtgccc
     7501 gcagttttta ttaaacataa cgtgggatct ccacgcgaat ctcgggtacg tgttccggac
     7561 atgggctctt ctccggtagc ggcggagctt ctacatccga gccctgctcc catgcctcca
     7621 gcgactcatg gtcgctcggc agctccttgc tcctaacagt ggaggccaga cttaggcaca
     7681 gcacgatgcc caccaccacc agtgtgccgc acaaggccgt ggcggtaggg tatgtgtctg
     7741 aaaatgagct cggggagcgg gcttgcaccg ctgacgcatt tggaagactt aaggcagcgg
     7801 cagaagaaga tgcaggcagc tgagttgttg tgttctgata agagtcagag gtaactcccg
     7861 ttgcggtgct gttaacggtg gagggcagtg tagtctgagc agtactcgtt gctgccgcgc
     7921 gcgccaccag acataatagc tgacagacta acagactgtt cctttccatg ggtcttttct
     7981 gcagtcaccg tccttgacac gaagcttccc ggccacagca atcgatccat gacacaacca
     8041 gtgacaagca gtcccccagt gaaaccgcat ccgca
//
