LOCUS       sequence-381949-        7666 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV overexpression of ChemoG-NAD in the nucleus of mammalian cell 
            lines.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7666)
  AUTHORS   Hellweg L, Edenhofer A, Barck L, Huppertz MC, Frei MS, Tarnawski M, 
            Bergner A, Koch B, Johnsson K, Hiblot J
  TITLE     A general method for the development of multicolor biosensors with 
            large dynamic ranges.
  JOURNAL   Nat Chem Biol. 2023 Jun 8. doi: 10.1038/s41589-023-01350-1.
  PUBMED    37291200
REFERENCE   2  (bases 1 to 7666)
  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 Chem 
            Biol. 2023 Jun 8. doi: 10.1038/s41589-023-01350-1."
COMMENT     Sequence Label: pCDNA5/FRT-ChemoG-NAD-NLS3x
FEATURES             Location/Qualifiers
     source          1..7666
                     /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     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(917..919,920..922,923..1435)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(962..983)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1223..1242)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1570..1591
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3497..3517
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3521..3541
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3545..3565
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3618..3635)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3624..3848
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3981..4000)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4132..4179
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             4187..5209
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYKSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5339..5460
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5376..5395)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5430..5449
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5509..5525)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5509..5525)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5522..5544)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5533..5549
                     /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(5557..5563,5564..5581,5582..5587))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5602..5623
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5740..5757)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5911..6499)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5991..6010)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6670..7461,7462..7530))
                     /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     7293..7312
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7531..7635)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccggtat 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 accaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      961 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     1021 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     1081 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     1141 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     1201 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     1261 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1321 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     1381 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     1441 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1501 gctgcccgac aaccactacc tgagcaccca gtccaaactg agcaaagacc ccaacgagaa
     1561 gcgcgatcac atggtcctgc tggagttcgt gcgcgccgcc gggatcactg gtggcaccat
     1621 gacactggaa gaagctcgta aacgtgtaaa cgaacttcgt gatctgatcc gttatcacaa
     1681 ttaccggtac tacgtgttgg ccgaccccga gattagcgat gcagagtacg atcgtctgct
     1741 gcgcgaactg aaggagttgg aggaacgttt tcctgagctc aaatcgccgg attcaccgac
     1801 attacaagtg ggcgcacggc cgctggaagc cacgttccgt cccgtgcgcc atccgacacg
     1861 catgtacagt ctggataacg cattcaactt ggatgaactg aaagcctttg aggaacgtat
     1921 cgaacgcgcg ctcgggcgca aaggtccgtt cgcctataca gttgaacatc tagtggatgg
     1981 cctgagcgtc aacttatatt acgaagaagg agtgttggtt tacggcgcaa cgcgtggcga
     2041 cggtgaagtg ggtgaggaag ttactcaaaa tttactcacc atcccgacta ttccacgccg
     2101 tctgaaaggt gtcccggaac gcctcgaggt gcgtggcgaa gtgtacatgc cgattgaggc
     2161 gtttctgcgc ctgaatgaag aactggagga acgtggcgag cgcattttta aaaatccgcg
     2221 taacgctgca gcaggcagct tgcgccagaa agatccgcgc attacggcta agcgtggttt
     2281 gcgcgcaacc ttctgggcgt taggtctggg tctcgaagag gtggagcgtg aaggagtagc
     2341 gactcaattt gcgttgctgc attggctcaa agagaaaggg ttcccggtcg aacacggata
     2401 tgcacgtgcc gttggggcgg aaggcgtgga ggcggtttat caggactggc tgaaaaaacg
     2461 ccgtgcgctg ccgtttgagg cgaatggagt tgcggtcaaa ctggatgaac ttgccctgtg
     2521 gcgtgaactg ggatacaccg cgcgtgcacc gcgttttgcc attgcgtaca agttcccttc
     2581 tggcggtatc ggtactggct ttccattcga cccccattat gtggaagtcc tgggcgagcg
     2641 catgcactac gtcgatgttg gtccgcgcga tggcacccct gtgctgttcc tgcacggtaa
     2701 cccgacctcc tcctacgtgt ggcgcaacat catcccgcat gttgcaccga cccatcgctg
     2761 cattgctcca gacctgatcg gtatgggcaa atccgacaaa ccagacctgg gttatttctt
     2821 cgacgaccac gtccgcttca tggatgcctt catcgaagcc ctgggtctgg aagaggtcgt
     2881 cctggtcatt cacgactggg gctccgctct gggtttccac tgggccaagc gcaatccaga
     2941 gcgcgtcaaa ggtattgcat ttatggagtt catccgccct atcccgacct gggacgaatg
     3001 gccagaattt gcccgcgaga ccttccaggc cttccgcacc accgacgtcg gccgcaagct
     3061 gatcatcgat cagaacgttt ttatcgaggg tacgctgccg atgggtgtcg tccgcccgct
     3121 gactgaagtc gagatggacc attaccgcga gccgttcctg aatcctgttg accgcgagcc
     3181 actgtggcgc ttcccaaacg agctgccaat cgccggtgag ccagcgaaca tcgtcgcgct
     3241 ggtcgaagaa tacatggact ggctgcacca gtcccctgtc ccgaagctgc tgttctgggg
     3301 caccccaggc gttctgatcc caccggccga agccgctcgc ctggccaaaa gcctgcctaa
     3361 ctgcaaggct gtggacatcg gcccgggtga aaatctgctg caagaagaca acccggacct
     3421 gatcggcagc gagatcgcgc gctggctgtc tactctggag atttccggta agtccggact
     3481 cagatctcga gctgatccaa aaaagaagag aaaggtagat ccaaaaaaga agagaaaggt
     3541 agatccaaaa aagaagagaa aggtaggatc caccggatct agatagtcta gagggcccgt
     3601 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     3661 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3721 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3781 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3841 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     3901 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3961 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4021 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4081 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtacct agaagttcct
     4141 attccgaagt tcctattctc tagaaagtat aggaacttcc ttggccaaaa agcctgaact
     4201 caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat
     4261 gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata
     4321 tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca
     4381 ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggaat tcagcgagag
     4441 cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac
     4501 cgaactgccc gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga
     4561 tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac
     4621 atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat
     4681 ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga
     4741 ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac
     4801 ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca
     4861 atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac
     4921 gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat
     4981 gctccgcatt ggtcttgacc aactctataa gagcttggtt gacggcaatt tcgatgatgc
     5041 agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg
     5101 tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc
     5161 cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaatagc acgtactacg
     5221 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5281 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     5341 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5401 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5461 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5521 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5581 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5641 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5701 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5761 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5821 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5881 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5941 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6001 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6061 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6121 gtatctcagt tcggtgcagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6181 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6241 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6301 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6361 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6421 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6481 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6541 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6601 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6661 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6721 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6781 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6841 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6901 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6961 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7021 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7081 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7141 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7201 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7261 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7321 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7381 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7441 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7501 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7561 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7621 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
