LOCUS       sequence-398148-        4749 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacteria protein expression of pnRFP biosensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4749)
  AUTHORS   Pang Y, Huang M, Fan Y, Yeh HW, Xiong Y, Ng HL, Ai HW
  TITLE     Development, Characterization, and Structural Analysis of a 
            Genetically Encoded Red Fluorescent Peroxynitrite Biosensor.
  JOURNAL   ACS Chem Biol. 2023 Jun 16;18(6):1388-1397. doi: 
            10.1021/acschembio.3c00139. Epub 2023 May 15.
  PUBMED    37185019
REFERENCE   2  (bases 1 to 4749)
  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"; doi: 
            "10.1021/acschembio.3c00139"; journalName: "ACS Chem Biol"; date: 
            "2023-06-16- 16"; volume: "18"; issue: "6"; pages: "1388-1397"
COMMENT     Sequence Label: pBAD-pnRFP
FEATURES             Location/Qualifiers
     source          1..4749
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /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     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     primer_bind     1044..1063
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1093..1110
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1150..1167)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1320..1406
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1498..1525
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1544..1635
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1636..1704,1705..2496)
                     /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(1854..1873)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2667..3255
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3156..3175
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3409..3426
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3441..3581
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3507..3526)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3667..3689
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3845..4723)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat gggctcgaga ataggtcggc tgggctcacc cgtagtttcc
      361 gagcggatgt accccgagga cggcgccctg aagagcgaga tcaagtaggg gctgaggctg
      421 aaggacggcg gccactacgc cgccgaggtc aagaccacct acaaggccaa gaagcccgtg
      481 cagctgcccg gcgcctacat cgtcgacatc aagttggaca tcgtgtccca caacgaggac
      541 tacaccatcg tggaacagtg cgaacgcgcc gagggccgcc actccactgg cggcatggac
      601 gagctgtaca agggaggtac aggcgggagt ctggtgagca agggcgagga ggataacatg
      661 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
      721 aagttcgaga tcgagggcga gggcgagggc cgcccctacg aggcctttca gaccgctaag
      781 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
      841 atgtacggct ccaaggccta cattaagcac ccagccgaca tccccgacta cttcaagctg
      901 tccttccccg agggcttcag gtgggagcgc gtggtgaact tcgaggacgg cggcattatt
      961 cacgttaacc aggactcctc cctgcaggac ggcgtattca tctacaaggt gaagctgcgc
     1021 ggcaccaact tcccccccga cggccccgta atgcagaaga agaccatggg ctgggaggct
     1081 acgcgtgacc aacaccatca ccaccatcat taagcttggc tgttttggcg gatgagagaa
     1141 gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt
     1201 gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg
     1261 ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc
     1321 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     1381 tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac
     1441 ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga
     1501 aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttttgttt atttttctaa
     1561 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     1621 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     1681 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     1741 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     1801 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     1861 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     1921 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     1981 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     2041 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     2101 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     2161 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     2221 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     2281 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     2341 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     2401 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     2461 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     2521 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     2581 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     2641 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2701 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2761 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     2821 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2881 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2941 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     3001 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     3061 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     3121 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     3181 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     3241 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     3301 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     3361 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     3421 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3481 tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     3541 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
     3601 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     3661 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     3721 agcagatcaa ttcgcgcgcg aaggcgaagc ggcatgcata atgtgcctgt caaatggacg
     3781 aagcagggat tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc tgattcgtta
     3841 ccaattatga caacttgacg gctacatcat tcactttttc ttcacaaccg gcacggaact
     3901 cgctcgggct ggccccggtg cattttttaa atacccgcga gaaatagagt tgatcgtcaa
     3961 aaccaacatt gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg
     4021 cctggctgat acgttggtcc tcgcgccagc ttaagacgct aatccctaac tgctggcgga
     4081 aaagatgtga cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa
     4141 aattgctgtc tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc cgattatcca
     4201 tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca
     4261 gatttatcgc cagcagctcc gaatagcgcc cttccccttg cccggcgtta atgatttgcc
     4321 caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg
     4381 caaatattga cggccagtta agccattcat gccagtaggc gcgcggacga aagtaaaccc
     4441 actggtgata ccattcgcga gcctccggat gacgaccgta gtgatgaatc tctcctggcg
     4501 ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg tccctgattt ttcaccaccc
     4561 cctgaccgcg aatggtgaga ttgagaatat aacctttcat tcccagcggt cggtcgataa
     4621 aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa
     4681 acgagtatcc cggcagcagg ggatcatttt gcgcttcagc catacttttc atactcccgc
     4741 cattcagag
//
