LOCUS       sequence-375361-        5777 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Clostridium plasmid expressing NanoLuc reporter from moderate, 
            constitutive promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5777)
  AUTHORS   Rostain W, Zaplana T, Boutard M, Baum C, Tabuteau S, Sanitha M, 
            Ramya M, Guss A, Ettwiller L, Tolonen AC
  TITLE     Tuning of Gene Expression in Clostridium phytofermentans Using 
            Synthetic Promoters and CRISPRi.
  JOURNAL   ACS Synth Biol. 2022 Dec 16;11(12):4077-4088. doi: 
            10.1021/acssynbio.2c00385. Epub 2022 Nov 25.
  PUBMED    36427328
REFERENCE   2  (bases 1 to 5777)
  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/acssynbio.2c00385"; journalName: "ACS Synth Biol"; date: 
            "2022-12-16- 16"; volume: "11"; issue: "12"; pages: "4077-4088"
COMMENT     Sequence Label: pQnl_Pcphy23
FEATURES             Location/Qualifiers
     source          1..5777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(458..475)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(629..1217)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(709..728)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5584..5777,1..322))
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
ORIGIN
        1 tgttcggcgt aaccccgtcg attaccagtg tgccatagtg caggatcacc ttaaagtgat
       61 gatcatccac agggtacacc accttaaaaa ttttttcgat ctggcccatt tggtcgccgc
      121 tcagaccttc atacgggatg atgacatgga tgtcgatctt cagcccattt tcaccgctca
      181 ggacaatcct ttggatcgga gttacggaca ccccgagatt ctgaaacaaa ctggacacac
      241 ctccctgttc aaggacttgg tccaggttgt agccggctgt ctgtcgccag tccccaacga
      301 aatcttcgag tgtgaagacc ataaaaaacc tcctttgcct tatatactgt gttcgggaac
      361 tttagtattt tatttaaaat attaaatttt atttttatta atgaatcggc caacgcgcgg
      421 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
      481 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
      541 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
      601 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
      661 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
      721 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
      781 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
      841 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
      901 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
      961 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1021 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1081 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1141 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1201 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1261 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1321 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     1381 ctgacagtat ctaatcactt tgactagcaa atactaacaa caagacacac acaccaaaaa
     1441 tcaaaaattc actactttta gttaaaaacc acgtaaccac aagaactaat ccaatccatg
     1501 taatcgggtt cttcaaatat ttctccaaga ttttcctcct ctaatatgct caacttaaat
     1561 gacctattca ataaatctat tatgctgcta aatagtttat aggacaaata agtatactct
     1621 aatgacctat aaaagataga aaattaaaaa atcaagtgtt cgcttcgctc tcactgcccc
     1681 tcgacgtttt agtagccttt ccctcacttc gttcagtcca agccaactaa aagttttcgg
     1741 gctactctct ccttctcccc ctaataatta attaaaatct tactctgtat atttctgcta
     1801 atcattcgct aaacagcaaa gaaaaaacaa acacgtatca tagatataaa tgtaatggca
     1861 tagtgcgggt tttattttca gcctgtatca tagctaaaca aatcgagttg tgtgtccgtt
     1921 ttagggcgtt ctgctagctt gtttaaagtc tcttgaatga atgtatgctc taagtcaaaa
     1981 gaatttgtca gcgcctttat atagctttct ttttcttctt tttttacttt aatgatcgat
     2041 agcaacaatg atttaacact agcaagttga atgccaccat ttcttcctgg tttaatctta
     2101 aagaaaattt cctgattcgc cttcagtacc ttcagcaatt tatctaatgt ccgttcagga
     2161 atgcctagca cttctctaat ctcttttttg gtcgtcacta aataaggctt gtatacatcg
     2221 cttttttcgc taatataagc cattaaatct tctttccatt ctgacaaatg aacacgttga
     2281 cgttcgcttc tttttttctt gaatttaaac cacccttgac ggacaaataa atctttactg
     2341 gttaaatcac ttgataccca agctttgcaa agaatggtaa tgtattccct attagcccct
     2401 tgatagtttt ctgaataggc acttctaaca attttgatta cttctttttc ttctaagggt
     2461 tgatctaatc gattattaaa ctcaaacata ttatattcgc acgtttcgat tgaatagcct
     2521 gaactaaagt aggctaaaga gagggtaaac atgacgttat tacgccctat taaacccttt
     2581 tctcctgaaa atttcgtttc gtgcaataag agattaaacc agggttcatc tacttgtttt
     2641 ttgccttctg taccgcttaa aaccgttaga cttgaacgag taaagccctt attatctgtt
     2701 tgtttgaaag accaatcttg ccattctttg aaagaataac ggtaattagg atcaaaaaat
     2761 tctacattgt ccgttcttgg tatgcgagca ataccaaaat gattacacgt tagatcaact
     2821 ggcaaagact ttccaaaata ttctcggata ttttgcgaaa ttattttggc tgctttgaca
     2881 gatttaaatt ctgattttga agtcacatag actggcgttt ctaaaacaaa atatgcttga
     2941 taacctttat cagatttgat aatcatagta ggcataaaac ctaaatcaat agcggttgtt
     3001 aaaatatcgc ttgctgaaat agtttctttt gccgtgtgaa tatcaaaatc aataaagaag
     3061 gtattgattt gtcttaaatt gttttcagaa tgtcctttcg tgtatgaacg gttttcgtct
     3121 gcatacgttc cataacgata aacgttgggt gtccaatgtg taaatgtatc ttgattttct
     3181 tgaatcgctt cctcggaagt cagaacaaca ccacgaccgc caatcatgct tgattttgag
     3241 cgatacgcaa aaatagcccc tttgctttta cctggcttgg tagtgattga gcgaatttta
     3301 ctatttttaa atttgtactt taacaagccg tcatgaagca cagtttctac aacaaaaggg
     3361 atattcattc agctgttctc ctttcctata aatcctataa aataggttgt ttaattaact
     3421 tggtttgctt tttcattcaa ctgtttcaat attgcatgtt ttgaaaaaga tttttttcct
     3481 ttataagtca atttttttcc actaatcgaa taaattattt tgttattttc tattaactta
     3541 tatatataat cttccccctc cgaagaaaaa tacttatctg attttgtttc taagtagata
     3601 tttctctttt ctaactcttt cttaaacgtt tctagtgtat agatatttgc taattttctt
     3661 atctccaata aactattttt tatataagtt ttacattcat catgattcat acaaactcca
     3721 ccttctataa atgaatacaa aaaaagcaat caaacgattt ccgattgatt gcttaacaat
     3781 tcttaaattc agtagcttag atacttgaaa actctctgat ttccctatat aatgatagta
     3841 cggttatata ccgtcttcaa acaaagttaa ttaaataact tcttacgagg gaagagttca
     3901 tctgactaac tgataagcgt tggtttggca atcttatcgg gctatgcatt tataaaatgt
     3961 cgtcaaacat tttataaatg tttcatggct cttttttcgt ttctattcag ttcgttgttt
     4021 cgttatatct agtataccgc ttttaaaaaa aataagcaac gatttcgtgc attattcaca
     4081 cgaagtcatt gcttttttct tcttccattt ctaaatccaa tgttacttgt tctgattctg
     4141 tttctggttc tggttctgtt ggctcatttg ggattaaatc cactactagc gttgagttag
     4201 ttaactttgc aatttgttct agtgttttta tggttggatc tgattttcct gattctattc
     4261 gtgaataatt tgatctactc atttctaatt cttgggagaa tttttctttt gatttgccca
     4321 aattccccgt aggcgctagg gacctcttta gcttcttgga agctgtcagt agtatatcta
     4381 ataatttatc tccattccct ttagtaacgt gtaactttcc aaatttaaaa aagcgactca
     4441 tagaattatt tcctcccgtt aaataataga taactattaa aaatagacaa tacttgctca
     4501 taagtaatgg tacttaaatt gtttactttg gcgtgtttca ttgcttgatg aaactgattt
     4561 ttagtaaaca gttgacgata ttctcgattg acccattttg aaacaaagta cgtatatagc
     4621 ttccaatatt tatctggaac atctgtggta tggcgggtaa gttttattaa gacactgttt
     4681 acttttggtt taggatgaaa gcattccgct ggcagcttaa gcaattgctg aatcgagact
     4741 tgagtgtgca agagcaaccc tagtgttcgg tgaatatcca aggtacgctt gtagaatcct
     4801 tcttcaacaa tcagatagat gtcagacgca tggctttcaa aaaccacttt tttaataatt
     4861 tgtgtgctta aatggtaagg aatactccca acaattttat acctctgttt gttagggaat
     4921 tgaaactgta gaatatcttg gtgaattaaa gtgacacgaa tgttcagttt taatttttct
     4981 gacgataagt tgaatagatg actgtctaat tcaatagacg ttacctgttt acttatttta
     5041 gccagtttcg tcgttaaatg ccctttacct gttccaattt cgtaaacggt atcggtttct
     5101 tttaaattca attgttttat tatttggttg agtacttttt cactcgttaa aaagttttga
     5161 gaatatttta tatttttgtt catgtaatta ctcctgaagt gattacatct gtaaataaat
     5221 acagaagtta aacgatttgt ttgtaatttt agttatctgt ttaaaaagtc ataagattag
     5281 tcactggtag gaattaatct aacgtattta tttatctgcg taatcactgt ttttagtctg
     5341 tttcaaaaca gtagatgttt tatctacatt acgcatttgg aataccaaca tgacgaatcc
     5401 ctccttctta attacaaatt tttagcatct aatttaactt caattcctat tatacacaaa
     5461 attttaagat actgcactat caacacactc ttaagtttgc ttctaaggtc gacggattta
     5521 aaagaaaaaa agatggtgag attatctcac catctttttt attttactca atagaaataa
     5581 atattacgcc agaatgcgtt cgcacagccg ccagccggtc actccgttga tggttactcg
     5641 gaacagcagg gagccgtcgg ggttgatcag gcgctcgtcg ataattttgt tgccgttcca
     5701 cagggtccct gttacagtga tctttttgcc gtcgaacacg gcgatgcctt catacggccg
     5761 tccgaaatag tcgatca
//
