LOCUS       sequence-323577-        4818 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Bacterial expression of genetically encoded fluorescent indicator 
            for measuring NAD+/NADH ratio.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4818)
  AUTHORS   Bilan DS, Matlashov ME, Gorokhovatsky AY, Schultz C, Enikolopov G, 
            Belousov VV
  TITLE     Genetically encoded fluorescent indicator for imaging NAD/NADH ratio
            changes in different cellular compartments.
  JOURNAL   Biochim Biophys Acta. 2013 Nov 25. pii: S0304-4165(13)00519-9. doi: 
            10.1016/j.bbagen.2013.11.018.
  PUBMED    24286672
REFERENCE   2  (bases 1 to 4818)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biochim 
            Biophys Acta. 2013 Nov 25. pii: S0304-4165(13)00519-9. doi: 
            10.1016/j.bbagen.2013.11.018."
COMMENT     Sequence Label: pQE-30-RexYFP
FEATURES             Location/Qualifiers
     source          1..4818
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    30..46
                     /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)."
     RBS             65..76
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             95..112
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     619..640
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(749..770)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1010..1029)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     terminator      1565..1659
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1703..2362
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1769..1788)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2427..2513
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2560..2582)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2668..2808
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2723..2742
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2823..2840)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2994..3582)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3074..3093)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3753..4544,4545..4613))
                     /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     4376..4395
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4614..4718)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4786..4804
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaatttc acacagaatt
       61 cattaaagag gagaaattaa ctatgagagg atcgcatcac catcaccatc acggatccgc
      121 atgcgagctc ggtaccccgg gtcgacctgc aggcatgaag gtccccgaag cggccatcag
      181 ccgcctgatc acctacctcc gcatcctgga ggagctggag gcccaggggg ttcaccggac
      241 gagctccgag cagcttgggg agctcgccca ggtcacggcc ttccaggtgc ggaaggacct
      301 ttcctacttc ggctcctacg ggacgcgcgg ggtggggtac accgtccccg tcctgaagcg
      361 ggagctccgc cacatcctgg ggctcaaccg ctccgccggc tacaacagcg acaacgtcta
      421 tatcatggcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacgt
      481 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
      541 ccccgtgctg ctgcccgaca accactacct gagcttccag tccgtcctga gcaaagaccc
      601 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactcc
      661 cggcatggac gagctgaaca acgtggatgg cggtagcggt ggcaccggca gcaagggcga
      721 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
      781 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
      841 gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctcgg
      901 ctacggcctg aagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
      961 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     1021 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1081 gaagggcatc ggcttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacgg
     1141 caccaagtgg ggcctgtgca tcgtgggcat gggccggctc gggagcgccc tggccgacta
     1201 cccggggttc ggggagagct ttgagctcag gggcttcttt gacgtggacc cggagaaggt
     1261 gggccgcccg gtgcggggcg gggtcattga gcacgtggac ctcctgcccc agcgggttcc
     1321 cggccgcatt gagatcgccc tcctcaccgt gccccgggaa gcggcccaga aggcggcgga
     1381 cctgttggtg gcggcgggga tcaaggggat cctgaacttc gccccggtgg tcctcgaggt
     1441 gcccaaggag gtggcggtgg agaacgtgga cttcttggcg gggctcacgc ggctttcttt
     1501 cgctatcctg aaccccaagt ggcgggaaga gatgatgggg tgaaagctta attagctgag
     1561 cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga
     1621 acgctcggtt gccgccgggc gttttttatt ggtgagaatc caagctagct tggcgagatt
     1681 ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat
     1741 atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta
     1801 taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca
     1861 caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt
     1921 tcgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac
     1981 cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt
     2041 ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta
     2101 tttccctaaa gggtttattg agaatatgtt tttcgtctca gccaatccct gggtgagttt
     2161 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat
     2221 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca
     2281 tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga
     2341 tgagtggcag ggcggggcgt aattttttta aggcagttat tggtgccctt aaacgcctgg
     2401 ggtaatgact ctctagcttg aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     2461 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccct
     2521 ctagagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     2581 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     2641 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
     2701 agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     2761 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct
     2821 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2881 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2941 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3001 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3061 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3121 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3181 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3241 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3301 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3361 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3421 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3481 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3541 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3601 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3661 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3721 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3781 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3841 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3901 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3961 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4021 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4081 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4141 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4201 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4261 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4321 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4381 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4441 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4501 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4561 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4621 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4681 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4741 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     4801 aggccctttc gtcttcac
//
