LOCUS       sequence-464244-        8185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses NADPH/NADP+ biosensor NAPstar3b in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8185)
  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 8185)
  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: pcDNA3.1(+) NAPstar3b
FEATURES             Location/Qualifiers
     source          1..8185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3117..3135)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(3776..3793)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3782..4006
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4052..4480
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4139..4158)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4349..4370
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4489..4509)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4494..4823
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4674..4809
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4736..4755
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4890..5684
                     /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(4944..4963)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5554..5573
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5858..5979
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5895..5914)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5949..5968
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6028..6044)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6028..6044)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6041..6063)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6052..6068
                     /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(6076..6082,6083..6100,6101..6106))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6121..6142
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6259..6276)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6430..7018)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6510..6529)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7189..7980,7981..8049))
                     /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     7812..7831
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8050..8154)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgcca ccatgaaggt gccagaggcc gccatcagca ggctgatcac
      961 ctacctgcgc atcctggagg agctggaggc acagggagtg cacagaacag cctccgagca
     1021 gctgggagag ctggcccagg tgaccgcctt ccaggtggac aaggatctga gctactttgg
     1081 ctcctatggc accgacggcg tgggctatac agtgcccgtg ctgaagcggg agctgagaca
     1141 catcctgggc ctgaacagga agtggggact gtgcatcgtg ggaatgggcc gcctgggctc
     1201 tgccctggcc gattggccag gattcggaga gagctttgag ctgaggggct tcttttcccg
     1261 caaagccgaa aaagtgggca ggcccgtgcg cggaggctat atcgagcacg tggacctgct
     1321 gccacagcgg gtgcccggca gaatcgagat cgccctgctg acagtgccta gggaggcagc
     1381 acagaaggca gcagatctgc tggtggccgc cggcatcaag ggcatcctga acttcgcccc
     1441 agtggtgctg gaggtgccca aggaggtggc cgtggagaat gtggacatcc tggccggcct
     1501 gaccagactg agcttcgcca tcctgaaccc aacatggtcc gccgcaggag gacacggatt
     1561 taccgcacac aacgtgtaca tcatggccga taagcagaag aacggcatca aggccaactt
     1621 caagatcagg cacaacatcg aggacggagg agtgcagctg gccgatcact accagcagaa
     1681 caccccaatc ggagacggac ctgtgctgct gccagataat cactatctgt ccatccagtc
     1741 taagctgagc aaggacccta atgagaagcg cgatcacatg gtgctgctgg agttcgtgac
     1801 cgcagcagga atcacacacg gaatggacga gctgtacaag ggcggcacag gcggctccat
     1861 ggtgtctaag ggagaggagc tgttcaccgg agtggtgccc atcctggtgg agctggacgg
     1921 cgatgtgaac ggccacaagt ttagcgtgtc cggagaggga gagggagatg caacctatgg
     1981 caagctgaca ctgaagttca tctgcaccac aggcaagctg cccgtgcctt ggccaacact
     2041 ggtgaccaca ttctcctacg gcgtgatggt gtttgcccgc tatcccgacc acatgaagca
     2101 gcacgatttc tttaagtctg ccatgcctga gggctacgtg caggagcgga ccatcttctt
     2161 taaggacgat ggcaactata agacaagagc cgaggtgaag ttcgagggcg acaccctggt
     2221 gaataggatc gagctgaagg gcatcgactt taaggaggat ggcaacatcc tgggccacaa
     2281 gctggagtac aatacaaagg tgcctgaggc cgccatctct cggctgatca cctatctgag
     2341 aatcctggag gaactggagg cccagggcgt gcaccggacc gcctccgaac agctgggcga
     2401 gctggcccag gtcactgcct tccaggtgga cgaggatctg tcttactttg gcagctacgg
     2461 caccgatggc gtgggctaca cagtgccagt gctgaaaaga gaactgaggc acatcctggg
     2521 cctgaatcgg aagtggggcc tgtgcattgt cggaatgggc agactgggct ccgccctggc
     2581 cgattggcct ggattcggcg agtcttttga gctgagaggc ttcttcagcc ggaaagccga
     2641 aaaagtcggc aggccagtgc gcggaggagt cattgaacat tatgatctgc tgccacagag
     2701 ggtgcctgga aggattgaaa tcgccctgct gaccgtgccc agggaggcag cccagaaggc
     2761 cgccgacctg ctggtggctg ctggaattaa aggcatcctg aattttgccc ctgtggtgct
     2821 ggaggtgcca aaagaagtcg ctgtggagaa tgtggacttc ctggccggcc tgacaaggct
     2881 gagctttgcc atcctgaacc ctaagtggag ggaggagatg atgggctccg gcaccggagg
     2941 aaatgcctcc gacggaggcg gctctggcgg catggtcagc aagggcgagg aggataacat
     3001 ggccatcatc aaggagttca tgagatttaa ggtgcacatg gagggcagcg tgaatggcca
     3061 cgagtttgag atcgaaggag agggagaggg caggccttac gagggcaccc agacagccaa
     3121 gctgaaggtg accaagggag gaccactgcc tttcgcctgg gacatcctgt ccccacagtt
     3181 tatgtacggc tctaaggcct atgtgaagca cccagccgac atccccgatt atctgaagct
     3241 gagcttcccc gagggcttta agtgggagcg cgtgatgaac ttcgaggatg gcggcgtggt
     3301 gaccgtgaca caggacagct ccctgcagga tggcgagttc atctacaagg tgaagctgcg
     3361 gggcacaaat tttccttccg acggcccagt gatgcagaag aagaccatgg gctgggaggc
     3421 ctctagcgag cgcatgtatc ccgaggatgg cgccctgaag ggcgagatca agcagcggct
     3481 gaagctgaag gacggcggcc actacgatgc cgaggtgaag accacatata aggccaagaa
     3541 gccagtgcag ctgcctggag cctacaacgt gaatatcaag ctggacatca cctctcacaa
     3601 cgaggattac acaatcgtgg agcagtatga gagggcagag ggcagacaca gcaccggagg
     3661 catggacgag ctgtataagt gaggatccac tagtccagtg tggtggaatt ctgcagatat
     3721 ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     3781 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3841 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3901 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3961 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     4021 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     4081 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4141 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4201 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4261 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4321 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4381 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4441 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4501 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4561 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4621 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4681 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4741 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4801 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4861 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4921 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4981 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5041 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     5101 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5161 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5221 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5281 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5341 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5401 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5461 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5521 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5581 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5641 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5701 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5761 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5821 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5881 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5941 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     6001 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6061 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6121 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6181 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6241 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6301 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6361 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6421 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6481 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6541 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6601 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6661 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6721 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6781 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6841 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6901 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6961 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7021 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7081 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7141 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7201 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7261 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7321 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7381 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7441 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7501 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7561 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7621 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7681 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7741 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7801 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7861 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7921 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7981 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8041 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8101 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8161 tccccgaaaa gtgccacctg acgtc
//
