LOCUS       sequence-464311-        8307 bp DNA     circular SYN 07-MAR-2025
DEFINITION  Expresses NADPH/NADP+ biosensor NAPstar4 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 Mar 7, 2025 from SnapGene Server 7.0.3
            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"
FEATURES             Location/Qualifiers
     source          1..8307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    76..579
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        609..713
                     /gene="bla"
                     /label=AmpR promoter
     CDS             714..1574
                     /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(932..951)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1745..2333
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2234..2253
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2487..2504
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2621..2642
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2657..2687
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2695..2711
                     /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     2700..2722
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2719..2735
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2719..2735
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2754..2774
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2756..2774
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2792..3190
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      5975..6222
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(5991..6009)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(6240..6259)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6241..6259)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6269..6286)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6269..6285)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6278..6300)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6426..6881
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6513..6532)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6723..6744
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7141..7803)
                     /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(7804..7991)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(8138..8157)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8257..8279
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagggtcc ttttcatcac gtgctataaa aataattata atttaaattt tttaatataa
      121 atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt
      181 ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta tcttgctctt
      241 cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag accacacacg
      301 aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta tttaatctgc
      361 ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt tttaaacctt
      421 tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac tttctaaaat
      481 ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta ttccatcatt
      541 aaaagatacg aggcgcgtgt aagttacagg caagcgatcc gtcaggtggc acttttcggg
      601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      721 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      781 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      841 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      901 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
      961 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1021 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1081 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1141 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1201 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1261 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1321 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1381 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1441 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1501 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1561 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1621 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1681 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1741 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1801 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1861 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     1921 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1981 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2041 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2101 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2161 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2221 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2281 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2341 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2401 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2461 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2521 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     2581 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     2641 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     2701 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
     2761 accctcacta aagggaacaa aagctggagc tcatagcttc aaaatgtttc tactcctttt
     2821 ttactcttcc agattttctc ggactccgcg catcgccgta ccacttcaaa acacccaagc
     2881 acagcatact aaatttcccc tctttcttcc tctagggtgt cgttaattac ccgtactaaa
     2941 ggtttggaaa agaaaaaaga gaccgcctcg tttctttttc ttcgtcgaaa aaggcaataa
     3001 aaatttttat cacgtttctt tttcttgaaa attttttttt tgattttttt ctctttcgat
     3061 gacctcccat tgatatttaa gttaataaac ggtcttcaat ttctcaagtt tcagtttcat
     3121 ttttcttgtt ctattacaac tttttttact tcttgctcat tagaaagaaa gcatagcaat
     3181 ctaatctaag ttttctagaa ccatgggatc caaagtacca gaagccgcta tttccagatt
     3241 gataacatat ttgagaattt tagaagaatt agaagcacaa ggtgtccaca gaaccgcttc
     3301 tgaacaattg ggtgaattag ctcaagttac agcattccaa gttgataaag atttgtcata
     3361 ctttggttct tacggtactg atggtgttgg ttacacagtt ccagttttga agagagaatt
     3421 gagacatatc ttgggtttaa acagaaagtg gggtttatgt atcgttggta tgggtgactt
     3481 aggttctgct ttggcagatt ggccaggttt tggtgaatca tttgaattga gaggtttctt
     3541 ttcaagatca gctcaaaaag ttggtagacc agttagaggt ggtgttattg aacatgttga
     3601 tttgttgcca caaagagttc caggtagaat tgaaattgct ttgttaactg ttccaagaga
     3661 agctgcacaa aaagctgcag atttgttagt tgctgcaggt attaaaggta ttttaaactt
     3721 cgcaccagtt gttttggaag ttccaaaaga agttgctgtt gaaaacgttg atattttggc
     3781 aggtttaact agattgtctt ttgctatctt gaatccaaca tggtcagctg caggtggtca
     3841 tggttttact gctcataacg tttacatcat ggcagataag caaaagaatg gtattaaagc
     3901 taacttcaag atcagacata atattgaaga tggtggtgtt caattggcag atcattacca
     3961 acaaaacact ccaattggtg acggtccagt tttgttacca gataaccatt acttatctat
     4021 ccaatcaaaa ttgtctaaag atccaaacga aaagagagat catatggttt tgttagaatt
     4081 tgttactgct gcaggtatca cacatggtat ggatgaatta tacaaaggtg gtacaggtgg
     4141 ttcaatggtt tctaaaggtg aagaattgtt tactggtgtt gttccaattt tagttgaatt
     4201 ggatggtgac gttaatggtc ataaattttc tgtttcaggt gaaggtgaag gtgacgctac
     4261 ttacggtaaa ttgacattga agtttatttg tactactggt aaattaccag ttccttggcc
     4321 aacattggtt actacatttt cttacggtgt tatggttttt gctagatacc cagatcatat
     4381 gaagcaacat gatttcttta agtcagcaat gccagaaggt tacgttcaag aaagaactat
     4441 tttctttaaa gatgatggta actacaagac aagagctgag gttaagttcg aaggtgacac
     4501 tttagttaac agaatcgaat tgaaaggtat cgatttcaaa gaagatggta acatcttagg
     4561 tcacaaattg gaatataata caaaagttcc agaagctgct atttcaagat tgatcactta
     4621 cttaagaatt ttggaagaat tagaagctca aggtgttcat agaacagcat cagaacaatt
     4681 aggtgaatta gctcaagtca ctgcattcca agttgatgaa gatttgtcat acttcggttc
     4741 ttatggtact gacggtgtcg gttatacagt tccagtctta aaaagagaat tgagacacat
     4801 tttaggttta aatagaaaat ggggtttatg cattgtcggc atgggtgact taggttctgc
     4861 tttggcagac tggcctggtt tcggcgaatc atttgaattg cgtggtttct tttcaagatc
     4921 agcgcaaaag gtcggtagac cagttagagg tggtgttatt gagcacgttg atttgttacc
     4981 tcagagagtt ccaggcagaa tcgaaatcgc tttgttaaca gttccaagag aagccgctca
     5041 aaaggccgca gatttgttag ttgccgctgg tattaaaggt attttaaatt tcgctcctgt
     5101 cgttttggaa gttccaaaag aagttgcggt tgaaaacgtt gatttcttgg ctggtttaac
     5161 aagattgtct tttgcaatct tgaacccaaa gtggagagaa gaaatgatgg gttcaggtac
     5221 tggtggtaat gcttcagatg gtggtggttc tggtggtatg gtttcaaagg gtgaagaaga
     5281 taacatggca attattaagg agtttatgag attcaaagtt catatggaag gttctgttaa
     5341 cggtcatgaa tttgaaatcg aaggtgaagg tgaaggtaga ccatatgaag gtactcaaac
     5401 agctaaattg aaggttacta aaggtggtcc attaccattt gcatgggata ttttgtctcc
     5461 acaattcatg tacggttcaa aagcatatgt taagcatcca gcagatattc cagattactt
     5521 gaaattatct tttcctgaag gttttaaatg ggaaagagtt atgaacttcg aagatggtgg
     5581 tgttgttact gttacacaag attcttcatt acaagatggc gagtttatat ataaggttaa
     5641 attgagaggt acaaattttc catcagatgg tccagttatg caaaagaaaa ctatgggttg
     5701 ggaagcatct tctgaaagaa tgtacccaga agatggtgca ttgaagggtg aaattaaaca
     5761 aagattgaaa ttgaaggatg gtggtcatta cgatgctgaa gttaagacta catacaaggc
     5821 taagaaacca gttcaattac caggtgctta caacgttaac atcaaattgg atattacttc
     5881 tcataacgaa gattacacaa ttgttgaaca atatgaaaga gccgaaggta gacactcaac
     5941 aggtggtatg gatgaattgt ataagtaact cgagtcatgt aattagttat gtcacgctta
     6001 cattcacgcc ctccccccac atccgctcta accgaaaagg aaggagttag acaacctgaa
     6061 gtctaggtcc ctatttattt ttttatagtt atgttagtat taagaacgtt atttatattt
     6121 caaatttttc ttttttttct gtacagacgc gtgtacgcat gtaacattat actgaaaacc
     6181 ttgcttgaga aggttttggg acgctcgaag gctttaattt gcggccggta cccaattcgc
     6241 cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
     6301 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     6361 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     6421 aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     6481 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     6541 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     6601 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     6661 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     6721 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     6781 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     6841 atttaacgcg aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc
     6901 cttacgcatc tgtgcggtat ttcacaccgc atagatccgt cgagttcaag agaaaaaaaa
     6961 agaaaaagca aaaagaaaaa aggaaagcgc gcctcgttca gaatgacacg tatagaatga
     7021 tgcattacct tgtcatcttc agtatcatac tgttcgtata catacttact gacattcata
     7081 ggtatacata tatacacatg tatatatatc gtatgctgca gctttaaata atcggtgtca
     7141 ctacataaga acacctttgg tggagggaac atcgttggta ccattgggcg aggtggcttc
     7201 tcttatggca accgcaagag ccttgaacgc actctcacta cggtgatgat cattcttgcc
     7261 tcgcagacaa tcaacgtgga gggtaattct gctagcctct gcaaagcttt caagaaaatg
     7321 cgggatcatc tcgcaagaga gatctcctac tttctccctt tgcaaaccaa gttcgacaac
     7381 tgcgtacggc ctgttcgaaa gatctaccac cgctctggaa agtgcctcat ccaaaggcgc
     7441 aaatcctgat ccaaaccttt ttactccacg cacggcccct agggcctctt taaaagcttg
     7501 accgagagca atcccgcagt cttcagtggt gtgatggtcg tctatgtgta agtcaccaat
     7561 gcactcaacg attagcgacc agccggaatg cttggccaga gcatgtatca tatggtccag
     7621 aaaccctata cctgtgtgga cgttaatcac ttgcgattgt gtggcctgtt ctgctactgc
     7681 ttctgcctct ttttctggga agatcgagtg ctctatcgct aggggaccac cctttaaaga
     7741 gatcgcaatc tgaatcttgg tttcatttgt aatacgcttt actagggctt tctgctctgt
     7801 catctttgcc ttcgtttatc ttgcctgctc attttttagt atattcttcg aagaaatcac
     7861 attactttat ataatgtata attcattatg tgataatgcc aatcgctaag aaaaaaaaag
     7921 agtcatccgc taggtggaaa aaaaaaaatg aaaatcatta ccgaggcata aaaaaatata
     7981 gagtgtacta gaggaggcca agagtaatag aaaaagaaaa ttgcgggaaa ggactgtgtt
     8041 atgacttccc tgactaatgc cgtgttcaaa cgatacctgg cagtgactcc tagcgctcac
     8101 caagctctta aaacgggaat ttatggtgca ctctcagtac aatctgctct gatgccgcat
     8161 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     8221 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     8281 tttcaccgtc atcaccgaaa cgcgcga
//
