LOCUS       sequence-476334-        5342 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of red fluorescent pyruvate control biosensor 
            dApplePy1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5342)
  AUTHORS   Imai S, Hario S, Suerte C, Yamaguchi I, Sakoi K, Terai T, 
            Takahashi-Yamashiro K, Campbell RE
  TITLE     High-performance genetically-encoded green and red fluorescent 
            biosensors for pyruvate
  JOURNAL   bioRxiv 2025.04.17.649293
REFERENCE   2  (bases 1 to 5342)
  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: "bioRxiv 
            2025.04.17.649293"
COMMENT     Sequence Label: pBAD-dApplePy1
FEATURES             Location/Qualifiers
     source          1..5342
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     30..47
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    62..202
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(128..147)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     288..310
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(466..1344)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1371..1655
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1578..1597
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1701..1718
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1722..1754
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(1758..1766,1767..1781)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(3113..3130)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3283..3369
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3461..3488
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3507..3598
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3599..3667,3668..4459)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3817..3836)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4630..5218
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5119..5138
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
       61 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
      121 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
      181 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
      241 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
      301 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcagatca attcgcgcgc
      361 gaaggcgaag cggcatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac
      421 cctatgctac tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac
      481 ggctacatca ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt
      541 gcatttttta aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac
      601 ggtggcgata ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc
      661 ctcgcgccag cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga
      721 cggcgacaag caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg
      781 atcgctgatg tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc
      841 gttaatcgct tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc
      901 cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg
      961 cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt
     1021 aagccattca tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg
     1081 agcctccgga tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc
     1141 cggtcggcaa acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag
     1201 attgagaata taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt
     1261 ggcctcaatc ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag
     1321 gggatcattt tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa
     1381 ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac
     1441 caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac
     1501 aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca
     1561 cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg
     1621 acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc taacaggagg
     1681 aattaaccat ggggggttct catcatcatc atcatcatgg tatggctagc atgactggtg
     1741 gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatccgagc tcgagaatgg
     1801 tgcaatcttt ctccgatccg ctgctagaac tgctttccga tcacccggaa tcccaatatg
     1861 acttgctcga aacgcgccat gcgctggaag ggatcgcagc atattacgct gccttgtgca
     1921 gctcagatga agagaaagag aggatcagag agttgcatca tgcgattgag cttgcccaac
     1981 aaagtggtga tctcgatgcc gaaagtaacg ctgtacttca gtatcaaatc gcggtgaccg
     2041 aggcagcgca caatgtggtc cttttgcact tgctgagata tatggagcct atgctcgcac
     2101 agaatatccg acataacttt gagctgctgt actctggtac ggtggtttcc gagcggatgt
     2161 tccccgagga cggcgccctg aagagcgagg tcaagaaggg gctgaggctg aaggacggcg
     2221 gccactacgc cgtggaggtc aagaccacct acaaggccaa gaagcccgtg cccctgcccg
     2281 gcgcctacat cgtcgacatc atgttggaca tcgtgtccca caacgaggac tacactatcg
     2341 tggaacagtt cgaacgcgcc gagggccgcc accccaccgg cggcctggac gagctgtaca
     2401 agggaggtac aggcgggagt ctggtaagca agggcgagga ggatgacatg gccatcgtca
     2461 aggagttcat gcgcttcaag gtgcacatgg agggctccgt taacggccac gagttcgaga
     2521 tcgaaggcga gggcgagggc cgcccctacg aggcctttca gactgctaag ctgaaggtga
     2581 ccaagggtgg ccccctgccc ttcgcctggg acatcttatc ccctcagttc atgtacggct
     2641 ccaaggccta cattaagcac ccagccgata tccccgacta cttcaagctg tccttccccg
     2701 agggcttcag ctgggagcgc gtgatgtact tcgaggacgg cggcattatt cacgttaacc
     2761 aggactcctc cctgcaggac ggcgtattta tctacaaggt gaagctgcgc ggcaccaact
     2821 tcccccccga cggccccgta atgcagatga agaccatggg ctgggagtcg ggttggggac
     2881 gacgagagat gctgccgctc gtggcctccc atcgcactcg catatttgaa gctattatgg
     2941 caggaaaacc ggaggaagcc agggaagcgt cacatagaca cttggccttt attaaggaaa
     3001 tcctgctgga ccggtcaagg ggggaatcac gccgcgaact ctcattgaga aggctggagc
     3061 aaagagagaa ttgaaagctt ggctgttttg gcggatgaga gaagattttc agcctgatac
     3121 agattaaatc agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg
     3181 cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta
     3241 gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct
     3301 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     3361 aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg
     3421 gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat
     3481 ggcctttttg cgtttctaca aactcttttg tttatttttc taaatacatt caaatatgta
     3541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     3841 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4141 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaacaacc
     4681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4741 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     4801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5281 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     5341 ta
//
