LOCUS       sequence-36950-e        8045 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 8045)
  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 8045)
  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
FEATURES             Location/Qualifiers
     source          1..8045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(652..1263,1264..1284,1285..2019,2020..2022,2023..2652,
                     2653..2697)
                     /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             2698..3408
                     /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(2843..2861)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3111..3130
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3530..3664
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3567..3586)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3621..3640
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3688..3704)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3688..3704)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3701..3723)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3712..3728
                     /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(3736..3742,3743..3760,3761..3766))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3781..3802
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3919..3936)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4090..4678)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4170..4189)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4849..5640,5641..5709))
                     /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     5472..5491
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     5919..5937
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5975..5997)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6097..6116
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6348..6370
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6362..6379
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6363..6379
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        6928..7307
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7308..7511
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7462..7482
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7508..7532
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          join(7647..8045,1..428)
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
ORIGIN
        1 ggacatgggc tcttctccgg tagcggcgga gcttctacat ccgagccctg ctcccatgcc
       61 tccagcgact catggtcgct cggcagctcc ttgctcctaa cagtggaggc cagacttagg
      121 cacagcacga tgcccaccac caccagtgtg ccgcacaagg ccgtggcggt agggtatgtg
      181 tctgaaaatg agctcgggga gcgggcttgc accgctgacg catttggaag acttaaggca
      241 gcggcagaag aagatgcagg cagctgagtt gttgtgttct gataagagtc agaggtaact
      301 cccgttgcgg tgctgttaac ggtggagggc agtgtagtct gagcagtact cgttgctgcc
      361 gcgcgcgcca ccagacataa tagctgacag actaacagac tgttcctttc catgggtctt
      421 ttctgcagtc accgtccttg acacgaagct tcccggccac agcaatcgat ccatgacaca
      481 accagtgaca agcagtcccc cagtgaaacc gcatccgcat ccgagtccga taccgataaa
      541 gattctgaat cggagtgagt gccgcgtccg agagcgttcc ctgtccacgt ccaccatcgg
      601 cggagcaggt gtgcctgagg cctctagatt gggcccacca tggttgctag caaagttcct
      661 gaagcagcca tttccagact gattacttac ctgagaattc tggaagagct ggaagcccaa
      721 ggtgtacacc gcaccgcctc cgagcaactg ggcgagctgg cccaggtcac cgccttccag
      781 gttgataagg acctgtccta cttcggctcc tacggcactg acggcgttgg ctacactgta
      841 ccggtcctca agagagaact gcgccatatc ctcggtctga accgcaaatg gggcctgtgc
      901 atcgtgggga tgggccgcct gggatccgct cttgctgatt ggcctggttt tggcgagagc
      961 ttcgagctgc gcggtttctt cgatgttgac ccaggtatgg ttggtcggcc ggttcgcggt
     1021 ggtgttatcg aacacgttga tctgctgccc cagcgcgtac ctggtcgcat cgaaatcgct
     1081 ctgctgacgg ttccacgcga ggcagcacag aaggctgccg acctgctggt tgccgctggc
     1141 atcaagggca tcctgaactt cgctccagtt gtactggagg ttcccaagga agtggctgtt
     1201 gagaacgttg acatcctggc cggtctgacg cgtctgagct ttgccattct gaaccccacg
     1261 tggagcgcag caggtgggca tggtttcact gctcacaacg tgtatatcat ggctgacaaa
     1321 cagaagaacg gtatcaaggc caacttcaag atccgccaca acatcgaaga cggcggcgtt
     1381 cagctggctg atcactacca gcagaacacc ccaatcggcg acggccctgt tctgctgcct
     1441 gacaaccact acttaagcat ccagtctaaa ctgtccaaag acccaaacga gaagcgcgac
     1501 cacatggttc tgctggaatt tgttaccgcc gctggcatca cccacggcat ggacgaactg
     1561 tacaagggcg gcaccggggg atctatggtt tccaaaggcg aggaactgtt cactggtgtt
     1621 gttcctatcc tggttgaact ggacggcgac gttaacggcc acaaattcag cgttagcggg
     1681 gaaggcgaag gagacgctac ctacggcaag ctgactctga agttcatctg caccactggc
     1741 aagctgcctg ttccttggcc taccctggtt actaccttct cctatggtgt tatggttttc
     1801 gccagatatc ctgaccacat gaagcagcac gacttcttca agagcgccat gcctgagggc
     1861 tacgttcagg aacgcaccat ctttttcaag gacgacggca actataagac acgcgctgag
     1921 gttaagttcg agggcgacac tctggttaac cgcatcgagc tgaagggcat cgacttcaag
     1981 gaggacggca acatcctggg ccataagctt gaatataaca ccaaggtacc tgaagccgcc
     2041 atcagccgcc tcatcactta cctccgcatc ctggaggagc tcgaagccca aggcgttcac
     2101 cgcactgcaa gcgagcagct gggcgaactg gcacaggtta cagcattcca ggttgacgaa
     2161 gatctgtcct atttcggttc ctacggcacc gacggtgttg gctacacagt ccctgtcctg
     2221 aagcgcgagc tgcgccacat cctgggcctg aaccgcaagt ggggcttatg catcgttggc
     2281 atgggccgcc tcggtagtgc actggcagac tggccgggct ttggtgaatc tttcgaactg
     2341 cgcggcttct tcgacgttga tccaggcatg gttgggcgcc ctgttcgcgg tggcgttatc
     2401 gagcatgttg acctcctgcc acagcgcgtc ccgggtcgca tcgagatcgc cctgctgacc
     2461 gttcctcgcg aagctgcaca gaaggcagcc gacttactag tcgccgctgg tatcaaggga
     2521 attctgaact tcgcccctgt tgttctggag gttcctaagg aagttgctgt tgaaaacgtt
     2581 gatttcctgg ccggcctgac ccgcctgagc ttcgcaatcc tgaatcctaa gtggcgcgaa
     2641 gaaatgatgg gttccggaac tgggggcaac gcctctgacg gtggtgggtc tggtggtatg
     2701 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
     2761 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     2821 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
     2881 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc
     2941 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     3001 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     3061 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     3121 cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
     3181 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
     3241 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac
     3301 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     3361 gccgagggcc gccactccac cggcggcatg gacgaactgt acaagtgagt cgacgaattc
     3421 attgatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     3481 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     3541 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3601 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3661 gatccgtacc gagctcgcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     3721 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3781 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3841 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3901 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3961 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4021 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4081 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4141 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4201 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4261 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     4321 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4381 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4441 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4501 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     4561 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4621 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4681 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4741 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4801 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4861 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4921 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4981 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5041 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5101 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5161 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5221 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5281 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5341 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5401 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5461 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5521 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5581 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5641 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5701 aatactcata ctcttccttt ttcaataagc ggccgcggcc atgccggcca ctagtctcga
     5761 gttattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5821 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5881 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     5941 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     6001 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     6061 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     6121 gagtgcacca tatgaagacg tcgcctcctc actacttctg gaatagctca gaggccgagg
     6181 cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcg cgccattcgc cattcaggct
     6241 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     6301 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     6361 ttgtaaaacg acggccagtg ccatctgcag tgaataataa aatgtgtgtt tgtccgaaat
     6421 acgcgtttga gatttctgtc ccgactaaat tcatgtcgcg cgatagtggt gtttatcgcc
     6481 gatagagatg gcgatattgg aaaaatcgat atttgaaaat atggcatatt gaaaatgtcg
     6541 ccgatgtgag tttctgtgta actgatatcg ccatttttcc aaaagttgat ttttgggcat
     6601 acgcgatatc tggcgatacg cttatatcgt ttacggggga tggcgataga cgcctttggt
     6661 gacttgggcg attctgtgtg tcgcaaatat cgcagtttcg atataggtga cagacgatat
     6721 gaggctatat cgccgataga ggcgacatca agctggcaca tggccaatgc atatcgatct
     6781 atacattgaa tcaatattgg ccattagcca tattattcat tggttatata gcataaatca
     6841 atattggcta ttggccattg catacgttgt atccatatca taatatgtac atttatattg
     6901 gctcatgtcc aacattaccg ccatgttgac attgattatt gactagttat taatagtaat
     6961 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     7021 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     7081 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     7141 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     7201 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     7261 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     7321 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     7381 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     7441 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     7501 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
     7561 acctccatag aagacaccgg gaccgatcca gcctccgcgg ccgggaacgg tgcattggaa
     7621 cgcggattcc ccgtgccaag agtgacgtaa gtaccgccta tagagtctat aggcccaccc
     7681 ccttggcttc ttatgcatgc tatactgttt ttggcttggg gtctatacac ccccgcttcc
     7741 tcatgttata ggtgatggta tagcttagcc tataggtgtg ggttattgac cattattgac
     7801 cactccccta ttggtgacga tactttccat tactaatcca taacatggct ctttgccaca
     7861 actctcttta ttggctatat gccaatacac tgtccttcag agactgacac ggactctgta
     7921 tttttacagg atggggtctc atttattatt tacaaattca catatacaac accaccgtcc
     7981 ccagtgcccg cagtttttat taaacataac gtgggatctc cacgcgaatc tcgggtacgt
     8041 gttcc
//
