LOCUS       sequence-464314-        8307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NADPH/NADP+ biosensor NAPstar7 in S. cerevisiae..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8307)
  AUTHORS   Scherschel M, Niemeier JO, Jacobs LJHC, Hoffmann MDA, Diederich A, 
            Bell C, Hohne P, Raetz S, Kroll JB, Steinbeck J, Lichtenauer S, 
            Multhoff J, Zimmermann J, Sadhanasatish T, Rothemann RA, Grashoff C,
            Messens J, Ampofo E, Laschke MW, Riemer J, Roma LP, Schwarzlander M,
            Morgan B
  TITLE     A family of NADPH/NADP(+) biosensors reveals in vivo dynamics of 
            central redox metabolism across eukaryotes.
  JOURNAL   Nat Commun. 2024 Dec 19;15(1):10704. doi: 
            10.1038/s41467-024-55302-x.
  PUBMED    39702652
REFERENCE   2  (bases 1 to 8307)
  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 
            Commun."; date: "2024-12-19"; volume: "15(1):10704. doi"; pages: " 
            10.1038/s41467-024-55302-x"
COMMENT     Sequence Label: p413 TEF NAPstar7
FEATURES             Location/Qualifiers
     source          1..8307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1951..2198
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1967..1985)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2216..2235)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2217..2235)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2245..2262)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2245..2261)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2254..2276)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2402..2857
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2489..2508)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2699..2720
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3117..3779)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(3780..3967)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(4114..4133)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4233..4255
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4293..4311)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4359..4862
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4892..4996
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4997..5065,5066..5857)
                     /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     complement(5215..5234)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6028..6616
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6517..6536
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6770..6787
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6904..6925
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6940..6945,6946..6963,6964..6970)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6978..6994
                     /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)."
     primer_bind     6983..7005
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7002..7018
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7002..7018
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7037..7057
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7039..7057
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7075..7473
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 tcaaaattgt ctaaagatcc aaacgaaaag agagatcata tggttttgtt agaatttgtt
       61 actgctgcag gtatcacaca tggtatggat gaattataca aaggtggtac aggtggttca
      121 atggtttcta aaggtgaaga attgtttact ggtgttgttc caattttagt tgaattggat
      181 ggtgacgtta atggtcataa attttctgtt tcaggtgaag gtgaaggtga cgctacttac
      241 ggtaaattga cattgaagtt tatttgtact actggtaaat taccagttcc ttggccaaca
      301 ttggttacta cattttctta cggtgttatg gtttttgcta gatacccaga tcatatgaag
      361 caacatgatt tctttaagtc agcaatgcca gaaggttacg ttcaagaaag aactattttc
      421 tttaaagatg atggtaacta caagacaaga gctgaggtta agttcgaagg tgacacttta
      481 gttaacagaa tcgaattgaa aggtatcgat ttcaaagaag atggtaacat cttaggtcac
      541 aaattggaat ataatacaaa agttccagaa gctgctattt caagattgat cacttactta
      601 agaattttgg aagaattaga agctcaaggt gttcatagaa cagcatcaga acaattaggt
      661 gaattagctc aagtcactgc attccaagtt gatgaagatt tgtcatactt cggttcttat
      721 ggtactgacg gtgtcggtta tacagttcca gtcttaaaaa gagaattgag acacatttta
      781 ggtttaaata gaaaatgggg tttatgcatt gtcggcatgg gtagattagg ttctgctttg
      841 gcagactggc ctggtttcgg cgaatcattt gaattgcgtg gtttcttttc aagaaaggcg
      901 gaaaaggtcg gtagaccagt tagaggtggt gttattgagc acgttgattt gttacctcag
      961 agagttccag gcagaatcga aatcgctttg ttaacaactc caagagaagc cgctcaaaag
     1021 gccgcagatt tgttagttgc cgctggtatt aaaggtattt taaatttcgc tcctgtcgtt
     1081 ttggaagttc caaaagaagt tgcggttgaa aacgttgatt tcttggctgg tttaacaaga
     1141 ttgtcttttg caatcttgaa cccaaagtgg agagaagaaa tgatgggttc aggtactggt
     1201 ggtaatgctt cagatggtgg tggttctggt ggtatggttt caaagggtga agaagataac
     1261 atggcaatta ttaaggagtt tatgagattc aaagttcata tggaaggttc tgttaacggt
     1321 catgaatttg aaatcgaagg tgaaggtgaa ggtagaccat atgaaggtac tcaaacagct
     1381 aaattgaagg ttactaaagg tggtccatta ccatttgcat gggatatttt gtctccacaa
     1441 ttcatgtacg gttcaaaagc atatgttaag catccagcag atattccaga ttacttgaaa
     1501 ttatcttttc ctgaaggttt taaatgggaa agagttatga acttcgaaga tggtggtgtt
     1561 gttactgtta cacaagattc ttcattacaa gatggcgagt ttatatataa ggttaaattg
     1621 agaggtacaa attttccatc agatggtcca gttatgcaaa agaaaactat gggttgggaa
     1681 gcatcttctg aaagaatgta cccagaagat ggtgcattga agggtgaaat taaacaaaga
     1741 ttgaaattga aggatggtgg tcattacgat gctgaagtta agactacata caaggctaag
     1801 aaaccagttc aattaccagg tgcttacaac gttaacatca aattggatat tacttctcat
     1861 aacgaagatt acacaattgt tgaacaatat gaaagagccg aaggtagaca ctcaacaggt
     1921 ggtatggatg aattgtataa gtaactcgag tcatgtaatt agttatgtca cgcttacatt
     1981 cacgccctcc ccccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct
     2041 aggtccctat ttattttttt atagttatgt tagtattaag aacgttattt atatttcaaa
     2101 tttttctttt ttttctgtac agacgcgtgt acgcatgtaa cattatactg aaaaccttgc
     2161 ttgagaaggt tttgggacgc tcgaaggctt taatttgcgg ccggtaccca attcgcccta
     2221 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa
     2281 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     2341 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     2401 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2461 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2521 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2581 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2641 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2701 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2761 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2821 aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     2881 cgcatctgtg cggtatttca caccgcatag atccgtcgag ttcaagagaa aaaaaaagaa
     2941 aaagcaaaaa gaaaaaagga aagcgcgcct cgttcagaat gacacgtata gaatgatgca
     3001 ttaccttgtc atcttcagta tcatactgtt cgtatacata cttactgaca ttcataggta
     3061 tacatatata cacatgtata tatatcgtat gctgcagctt taaataatcg gtgtcactac
     3121 ataagaacac ctttggtgga gggaacatcg ttggtaccat tgggcgaggt ggcttctctt
     3181 atggcaaccg caagagcctt gaacgcactc tcactacggt gatgatcatt cttgcctcgc
     3241 agacaatcaa cgtggagggt aattctgcta gcctctgcaa agctttcaag aaaatgcggg
     3301 atcatctcgc aagagagatc tcctactttc tccctttgca aaccaagttc gacaactgcg
     3361 tacggcctgt tcgaaagatc taccaccgct ctggaaagtg cctcatccaa aggcgcaaat
     3421 cctgatccaa acctttttac tccacgcacg gcccctaggg cctctttaaa agcttgaccg
     3481 agagcaatcc cgcagtcttc agtggtgtga tggtcgtcta tgtgtaagtc accaatgcac
     3541 tcaacgatta gcgaccagcc ggaatgcttg gccagagcat gtatcatatg gtccagaaac
     3601 cctatacctg tgtggacgtt aatcacttgc gattgtgtgg cctgttctgc tactgcttct
     3661 gcctcttttt ctgggaagat cgagtgctct atcgctaggg gaccaccctt taaagagatc
     3721 gcaatctgaa tcttggtttc atttgtaata cgctttacta gggctttctg ctctgtcatc
     3781 tttgccttcg tttatcttgc ctgctcattt tttagtatat tcttcgaaga aatcacatta
     3841 ctttatataa tgtataattc attatgtgat aatgccaatc gctaagaaaa aaaaagagtc
     3901 atccgctagg tggaaaaaaa aaaatgaaaa tcattaccga ggcataaaaa aatatagagt
     3961 gtactagagg aggccaagag taatagaaaa agaaaattgc gggaaaggac tgtgttatga
     4021 cttccctgac taatgccgtg ttcaaacgat acctggcagt gactcctagc gctcaccaag
     4081 ctcttaaaac gggaatttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4141 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4201 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4261 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     4321 taatgtcatg ataataatgg tttcttaggg tccttttcat cacgtgctat aaaaataatt
     4381 ataatttaaa ttttttaata taaatatata aattaaaaat agaaagtaaa aaaagaaatt
     4441 aaagaaaaaa tagtttttgt tttccgaaga tgtaaaagac tctaggggga tcgccaacaa
     4501 atactacctt ttatcttgct cttcctgctc tcaggtatta atgccgaatt gtttcatctt
     4561 gtctgtgtag aagaccacac acgaaaatcc tgtgatttta cattttactt atcgttaatc
     4621 gaatgtatat ctatttaatc tgcttttctt gtctaataaa tatatatgta aagtacgctt
     4681 tttgttgaaa ttttttaaac ctttgtttat ttttttttct tcattccgta actcttctac
     4741 cttctttatt tactttctaa aatccaaata caaaacataa aaataaataa acacagagta
     4801 aattcccaaa ttattccatc attaaaagat acgaggcgcg tgtaagttac aggcaagcga
     4861 tccgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4921 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4981 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5041 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5101 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5161 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5221 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5281 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5341 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5401 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5461 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5521 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5581 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5641 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5701 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5761 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5821 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5881 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5941 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6001 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6061 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6121 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6181 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6241 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6301 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6361 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6421 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6481 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6541 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6601 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6661 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6721 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     6781 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     6841 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     6901 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     6961 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     7021 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctcatagc
     7081 ttcaaaatgt ttctactcct tttttactct tccagatttt ctcggactcc gcgcatcgcc
     7141 gtaccacttc aaaacaccca agcacagcat actaaatttc ccctctttct tcctctaggg
     7201 tgtcgttaat tacccgtact aaaggtttgg aaaagaaaaa agagaccgcc tcgtttcttt
     7261 ttcttcgtcg aaaaaggcaa taaaaatttt tatcacgttt ctttttcttg aaaatttttt
     7321 ttttgatttt tttctctttc gatgacctcc cattgatatt taagttaata aacggtcttc
     7381 aatttctcaa gtttcagttt catttttctt gttctattac aacttttttt acttcttgct
     7441 cattagaaag aaagcatagc aatctaatct aagttttcta gaaccatggg atccaaagta
     7501 ccagaagccg ctatttccag attgataaca tatttgagaa ttttagaaga attagaagca
     7561 caaggtgtcc acagaaccgc ttctgaacaa ttgggtgaat tagctcaagt tacagcattc
     7621 caagttgata aagatttgtc atactttggt tcttacggta ctgatggtgt tggttacaca
     7681 gttccagttt tgaagagaga attgagacat atcttgggtt taaacagaaa gtggggttta
     7741 tgtatcgttg gtatgggtag attaggttct gctttggcag attggccagg ttttggtgaa
     7801 tcatttgaat tgagaggttt cttttcaaga aaggctgaaa aagttggtag accagttaga
     7861 ggtggtgtta ttgaacatgt tgatttgttg ccacaaagag ttccaggtag aattgaaatt
     7921 gctttgttaa ctactccaag agaagctgca caaaaagctg cagatttgtt agttgctgca
     7981 ggtattaaag gtattttaaa cttcgcacca gttgttttgg aagttccaaa agaagttgct
     8041 gttgaaaacg ttgatatttt ggcaggttta actagattgt cttttgctat cttgaatcca
     8101 acatggtcag ctgcaggtgg tcatggtttt actgctcata acgtttacat catggcagat
     8161 aagcaaaaga atggtattaa agctaacttc aagatcagac ataatattga agatggtggt
     8221 gttcaattgg cagatcatta ccaacaaaac actccaattg gtgacggtcc agttttgtta
     8281 ccagataacc attacttatc tatccaa
//
