LOCUS       sequence-375202-        5469 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Clostridium expression vector (pBP1 origin, ermR) 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 5469)
  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 5469)
  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: pQmod2E-GG
FEATURES             Location/Qualifiers
     source          1..5469
                     /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      4679..5267
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5168..5187
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5421..5438
                     /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 ataactatgg tctcccgaat cttccgctag cataaaaata agaagcctgc atttgcaggc
     1261 ttcttatttt tatggcgcgc cgttctgaat ccttagctaa tggttcaaca ggtaactatg
     1321 acgaagatag caccctggat aagtctgtaa tggattctaa ggcatttaat gaagacgtgt
     1381 atataaaatg tgctaatgaa aaagaaaatg cgttaaaaga gcctaaaatg agttcaaatg
     1441 gttttgaaat tgattggtag tttaatttaa tatatttttt ctattggcta tctcgatacc
     1501 tatagaatct tctgttcact tttgtttttg aaatataaaa aggggctttt tagccccttt
     1561 tttttaaaac tccggaggag tttcttcatt cttgatacta tacgtaacta tttcgatttg
     1621 acttcattgt caattaagct agtaaaatca atggttaaaa aacaaaaaac ttgcattttt
     1681 ctacctagta atttataatt ttaagtgtcg agtttaaaag tataatttac caggaaagga
     1741 gcaagttttt taataaggaa aaatttttcc ttttaaaatt ctatttcgtt atatgactaa
     1801 ttataatcaa aaaaatgaaa ataaacaaga ggtaaaaact gctttagaga aatgtactga
     1861 taaaaaaaga aaaaatccta gatttacgtc atacatagca cctttaacta ctaagaaaaa
     1921 tattgaaagg acttccactt gtggagatta tttgtttatg ttgagtgatg cagacttaga
     1981 acattttaaa ttacataaag gtaatttttg cggtaataga ttttgtccaa tgtgtagttg
     2041 gcgacttgct tgtaaggata gtttagaaat atctattctt atggagcatt taagaaaaga
     2101 agaaaataaa gagtttatat ttttaactct tacaactcca aatgtaaaaa gttatgatct
     2161 taattattct attaaacaat ataataaatc ttttaaaaaa ttaatggagc gtaaggaagt
     2221 taaggatata actaaaggtt atataagaaa attagaagta acttaccaaa aggaaaaata
     2281 cataacaaag gatttatgga aaataaaaaa agattattat caaaaaaaag gacttgaaat
     2341 tggtgattta gaacctaatt ttgatactta taatcctcat tttcatgtag ttattgcagt
     2401 taataaaagt tattttacag ataaaaatta ttatataaat cgagaaagat ggttggaatt
     2461 atggaagttt gctactaagg atgattctat aactcaagtt gatgttagaa aagcaaaaat
     2521 taatgattat aaagaggttt acgaacttgc gaaatattca gctaaagaca ctgattattt
     2581 aatatcgagg ccagtatttg aaatttttta taaagcatta aaaggcaagc aggtattagt
     2641 ttttagtgga ttttttaaag atgcacacaa attgtacaag caaggaaaac ttgatgttta
     2701 taaaaagaaa gatgaaatta aatatgtcta tatagtttat tataattggt gcaaaaaaca
     2761 atatgaaaaa actagaataa gggaacttac ggaagatgaa aaagaagaat taaatcaaga
     2821 tttaatagat ggaatagaaa tagattaaag tgtaactata ctttatatat atatgattaa
     2881 aaaaataaaa aacaacagcc tattaggttg ttgtttttta ttttctttat taattttttt
     2941 aatttttagt ttttagttct tttttaaaat aagtttcagc ctctttttca atatttttta
     3001 aagaaggagt atttgcatga attgcctttt ttctaacaga cttaggaaat attttaacag
     3061 tatcttcttg cgccggtgat tttggaactt cataacttac taatttataa ttattatttt
     3121 cttttttaat tgtaacagtt gcaaaagaag ctgaacctgt tccttcaact agtttatcat
     3181 cttcaatata atattcttga cctatatagt ataaatatat ttttattata tttttacttt
     3241 tttctgaatc tattatttta taatcataaa aagttttacc accaaaagaa ggttgtactc
     3301 cttctggtcc aacatatttt tttactatat tatctaaata atttttggga actggtgttg
     3361 taatttgatt aatcgaacaa ccagttatac ttaaaggaat tataactata aaaatatata
     3421 ggattatctt tttaaatttc attattggcc tcctttttat taaatttatg ttaccataaa
     3481 aaggacataa cgggaatatg tagaatattt ttaatgtaga caaaatttta cataaatata
     3541 aagaaaggaa gtgtttgttt aaattttata gcaaactatc aaaaattagg gggataaaaa
     3601 tttatgaaaa aaaggttttc gatgttattt ttatgtttaa ctttaatagt ttgtggttta
     3661 tttacaaatt cggccggccg aagcaaactt aagagtgtgt tgatagtgca gtatcttaaa
     3721 attttgtata ataggaattg aagttaaatt agatgctaaa aatttgtaat taagaaggag
     3781 tgattacatg aacaaaaata taaaatattc tcaaaacttt ttaacgagtg aaaaagtact
     3841 caaccaaata ataaaacaat tgaatttaaa agaaaccgat accgtttacg aaattggaac
     3901 aggtaaaggg catttaacga cgaaactggc taaaataagt aaacaggtaa cgtctattga
     3961 attagacagt catctattca acttatcgtc agaaaaatta aaactgaata ctcgtgtcac
     4021 tttaattcac caagatattc tacagtttca attccctaac aaacagaggt ataaaattgt
     4081 tgggagtatt ccttaccatt taagcacaca aattattaaa aaagtggttt ttgaaagcca
     4141 tgcgtctgac atctatctga ttgttgaaga aggattctac aagcgtacct tggatattca
     4201 ccgaacacta gggttgctct tgcacactca agtctcgatt cagcaattgc ttaagctgcc
     4261 agcggaatgc tttcatccta aaccaaaagt aaacagtgtc ttaataaaac ttacccgcca
     4321 taccacagat gttccagata aatattggaa gctatatacg tactttgttt caaaatgggt
     4381 caatcgagaa tatcgtcaac tgtttactaa aaatcagttt catcaagcaa tgaaacacgc
     4441 caaagtaaac aatttaagta ccgttactta tgagcaagta ttgtctattt ttaatagtta
     4501 tctattattt aacgggagga aataattcta tgagtcgctt ttgtaaattt ggaaagttac
     4561 acgttactaa agggaatgtg tttaaactcc tttttgataa tctcatgacc aaaatccctt
     4621 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4681 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4741 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4801 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4861 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4921 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4981 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5041 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5101 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5161 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5221 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5281 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5341 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     5401 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     5461 gcagggccc
//
