LOCUS       sequence-375352-        5392 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Clostridium expression vector (pBP1 origin, cmR) with drop-out 
            Plac-RFP cassette flanked with BsaI sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5392)
  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 5392)
  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: pQmod2C-GG
FEATURES             Location/Qualifiers
     source          1..5392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     63..79
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    236..257
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(272..277,278..295,296..302)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    310..326
                     /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             344..355
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             362..1039
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1084..1155
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1171..1198
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      4602..5190
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5091..5110
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5344..5361
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cctgcaggat aaaaaaattg tagataaatt ttataaaata gttttatcta caattttttt
       61 atcaggaaac agctatgacc gcggccgctg tatccatatg accatgatta cgaattcgag
      121 ctcaggatga gacctcaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
      181 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
      241 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
      301 tgtgtggaat tgtgagcgga taacaatttc acacatacta gagaaagagg agaaatacta
      361 gatggcttcc tccgaagacg ttatcaaaga gttcatgcgt ttcaaagttc gtatggaagg
      421 ttccgttaac ggtcacgagt tcgaaatcga aggtgaaggt gaaggtcgtc cgtacgaagg
      481 tacccagacc gctaaactga aagttaccaa aggtggtccg ctgccgttcg cttgggacat
      541 cctgtccccg cagttccagt acggttccaa agcttacgtt aaacacccgg ctgacatccc
      601 ggactacctg aaactgtcct tcccggaagg tttcaaatgg gaacgtgtta tgaacttcga
      661 agacggtggt gttgttaccg ttacccagga ctcctccctg caagacggtg agttcatcta
      721 caaagttaaa ctgcgtggta ccaacttccc gtccgacggt ccggttatgc agaaaaaaac
      781 catgggttgg gaagcttcca ccgaacgtat gtacccggaa gacggtgctc tgaaaggtga
      841 aatcaaaatg cgtctgaaac tgaaagacgg tggtcactac gacgctgaag ttaaaaccac
      901 ctacatggct aaaaaaccgg ttcagctgcc gggtgcttac aaaaccgaca tcaaactgga
      961 catcacctcc cacaacgaag actacaccat cgttgaacag tacgaacgtg ctgaaggtcg
     1021 tcactccacc ggtgcttaat aacgctgata gtgctagtgt agatcgctac tagagccagg
     1081 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     1141 tcggtgaacg ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt
     1201 tataactatg gtctcccgaa tcttccgcta gcataaaaat aagaagcctg catttgcagg
     1261 cttcttattt ttatggcgcg ccgttctgaa tccttagcta atggttcaac aggtaactat
     1321 gacgaagata gcaccctgga taagtctgta atggattcta aggcatttaa tgaagacgtg
     1381 tatataaaat gtgctaatga aaaagaaaat gcgttaaaag agcctaaaat gagttcaaat
     1441 ggttttgaaa ttgattggta gtttaattta atatattttt tctattggct atctcgatac
     1501 ctatagaatc ttctgttcac ttttgttttt gaaatataaa aaggggcttt ttagcccctt
     1561 ttttttaaaa ctccggagga gtttcttcat tcttgatact atacgtaact atttcgattt
     1621 gacttcattg tcaattaagc tagtaaaatc aatggttaaa aaacaaaaaa cttgcatttt
     1681 tctacctagt aatttataat tttaagtgtc gagtttaaaa gtataattta ccaggaaagg
     1741 agcaagtttt ttaataagga aaaatttttc cttttaaaat tctatttcgt tatatgacta
     1801 attataatca aaaaaatgaa aataaacaag aggtaaaaac tgctttagag aaatgtactg
     1861 ataaaaaaag aaaaaatcct agatttacgt catacatagc acctttaact actaagaaaa
     1921 atattgaaag gacttccact tgtggagatt atttgtttat gttgagtgat gcagacttag
     1981 aacattttaa attacataaa ggtaattttt gcggtaatag attttgtcca atgtgtagtt
     2041 ggcgacttgc ttgtaaggat agtttagaaa tatctattct tatggagcat ttaagaaaag
     2101 aagaaaataa agagtttata tttttaactc ttacaactcc aaatgtaaaa agttatgatc
     2161 ttaattattc tattaaacaa tataataaat cttttaaaaa attaatggag cgtaaggaag
     2221 ttaaggatat aactaaaggt tatataagaa aattagaagt aacttaccaa aaggaaaaat
     2281 acataacaaa ggatttatgg aaaataaaaa aagattatta tcaaaaaaaa ggacttgaaa
     2341 ttggtgattt agaacctaat tttgatactt ataatcctca ttttcatgta gttattgcag
     2401 ttaataaaag ttattttaca gataaaaatt attatataaa tcgagaaaga tggttggaat
     2461 tatggaagtt tgctactaag gatgattcta taactcaagt tgatgttaga aaagcaaaaa
     2521 ttaatgatta taaagaggtt tacgaacttg cgaaatattc agctaaagac actgattatt
     2581 taatatcgag gccagtattt gaaatttttt ataaagcatt aaaaggcaag caggtattag
     2641 tttttagtgg attttttaaa gatgcacaca aattgtacaa gcaaggaaaa cttgatgttt
     2701 ataaaaagaa agatgaaatt aaatatgtct atatagttta ttataattgg tgcaaaaaac
     2761 aatatgaaaa aactagaata agggaactta cggaagatga aaaagaagaa ttaaatcaag
     2821 atttaataga tggaatagaa atagattaaa gtgtaactat actttatata tatatgatta
     2881 aaaaaataaa aaacaacagc ctattaggtt gttgtttttt attttcttta ttaatttttt
     2941 taatttttag tttttagttc ttttttaaaa taagtttcag cctctttttc aatatttttt
     3001 aaagaaggag tatttgcatg aattgccttt tttctaacag acttaggaaa tattttaaca
     3061 gtatcttctt gcgccggtga ttttggaact tcataactta ctaatttata attattattt
     3121 tcttttttaa ttgtaacagt tgcaaaagaa gctgaacctg ttccttcaac tagtttatca
     3181 tcttcaatat aatattcttg acctatatag tataaatata tttttattat atttttactt
     3241 ttttctgaat ctattatttt ataatcataa aaagttttac caccaaaaga aggttgtact
     3301 ccttctggtc caacatattt ttttactata ttatctaaat aatttttggg aactggtgtt
     3361 gtaatttgat taatcgaaca accagttata cttaaaggaa ttataactat aaaaatatat
     3421 aggattatct ttttaaattt cattattggc ctccttttta ttaaatttat gttaccataa
     3481 aaaggacata acgggaatat gtagaatatt tttaatgtag acaaaatttt acataaatat
     3541 aaagaaagga agtgtttgtt taaattttat agcaaactat caaaaattag ggggataaaa
     3601 atttatgaaa aaaaggtttt cgatgttatt tttatgttta actttaatag tttgtggttt
     3661 atttacaaat tcggccggcc agtgggcaag ttgaaaaatt cacaaaaatg tggtataata
     3721 tctttgttca ttagagcgat aaacttgaat ttgagaggga acttagatgg tatttgaaaa
     3781 aattgataaa aatagttgga acagaaaaga gtattttgac cactactttg caagtgtacc
     3841 ttgtacctac agcatgaccg ttaaagtgga tatcacacaa ataaaggaaa agggaatgaa
     3901 actatatcct gcaatgcttt attatattgc aatgattgta aaccgccatt cagagtttag
     3961 gacggcaatc aatcaagatg gtgaattggg gatatatgat gagatgatac caagctatac
     4021 aatatttcac aatgatactg aaacattttc cagcctttgg actgagtgta agtctgactt
     4081 taaatcattt ttagcagatt atgaaagtga tacgcaacgg tatggaaaca atcatagaat
     4141 ggaaggaaag ccaaatgctc cggaaaacat ttttaatgta tctatgatac cgtggtcaac
     4201 cttcgatggc tttaatctga atttgcagaa aggatatgat tatttgattc ctatttttac
     4261 tatggggaaa tattataaag aagataacaa aattatactt cctttggcaa ttcaagttca
     4321 tcacgcagta tgtgacggat ttcacatttg ccgttttgta aacgaattgc aggaattgat
     4381 aaatagttaa cttcaggttt gtctgtaact aaaaacaagt atttaagcaa aaacatcgta
     4441 gaaatacggt gttttttgtt accctaagtt tgggattttg gtcatgagat tatcaaaaag
     4501 gagtttaaac tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4561 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4621 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4681 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4741 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4801 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4861 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4921 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4981 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5041 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5101 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5161 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5221 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5281 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     5341 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcagggc cc
//
