LOCUS       sequence-375355-        6514 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Clostridium expression vector (pCD6 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 6514)
  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 6514)
  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: pQmod4E-GG
FEATURES             Location/Qualifiers
     source          1..6514
                     /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      5724..6312
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6213..6232
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6466..6483
                     /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 cccgccctta agtctaaaaa ttaggggaga tgtaaggatt
     1321 tgggaaaaat agaagatgtt ataatcataa atatggtatt cgtaggctta aagtcaaaaa
     1381 ggaggtgaaa tataaataga tttttagcta aattaagtaa gaaataggag gagatttatt
     1441 gaacaaaaaa ttagaaaaac catttgtata taagagagag tacgatttga ctggatatga
     1501 tgttgaaatt ttacaaaaat atgagttaga acaagcaata tatgtttatg ttgggagtag
     1561 ttgtgcatat aacatgagag ctagaagtag taaatggaga taccatataa gaacaaataa
     1621 taagtctata tgttgtaaca ttaaaaattt tatacataac ttggaattgt tttataaaat
     1681 ggaattaaag ttgtcagata atattattaa tgataagcta tactatagca atatagcaga
     1741 gtttgaagaa tttgaaacac tagaaaaagc tagagaggta gaaagtacta ttataagtca
     1801 atatcaattt ttagattcta taaatcacat gttaaaacaa aaaataattt tattgagtaa
     1861 taaggatagt gtgttaaaca taactaaaaa tggaaataca aattatttga aagtaaaaaa
     1921 taaatacata gaaaaacata agaacaagcc aataatgaga taccatatca actgtcaatt
     1981 caatacagat ggaagtgtca aaagtattac acaggagttt gaaccaatat tggaattaaa
     2041 caaaaaaaat accctaagcc gaccaagcag agtattttta aaataatatt ttaagataac
     2101 aacaaaatga gataatacta ctagacaatg acaactcaac taccaattga gtttatggag
     2161 ctaccaactc caatatcggt ctaactgatt aagtatctgt agttatataa taatattgct
     2221 atcaatttta gcatcttaac aatattatta tacatactaa gctaaaatta ttcaatagtt
     2281 gtaaaagttg attagtcaat aagtatatat ttaatgtagt gttatctctt aaaaaaacta
     2341 gataaggaga taataaatat atggaacaat tagattcaaa atataagttg aaaaaatttc
     2401 taatggcagt atttagagat ggtataggac aaggaaataa tcttattgat aatgaatatg
     2461 ttagagtatt tcaaaataat aaaagtaata gtaaacaatt agaactcgga gaagaattta
     2521 aagaatatag taaaacaact ttttttaaaa atatagatga tatagtagaa tttaccttcg
     2581 caaaaaatat ttattatgaa aatacatttt ttaacctatg tactactgat ggaaaagcag
     2641 gaaccaatga aaacttaata aatagatatg cattaggatt tgattttgac aaaaaagaat
     2701 taggacaagg ttttaattat aaagatataa ttaatttatt tactaagata ggattacatt
     2761 atcatatcct agttgatagt ggaaatggat tccatgttta tgtgctaatt aataaaacta
     2821 ataacattaa gttagtatca gaagttacaa atacattaat aaataaattg ggtgcagata
     2881 aacaagcaaa tttatctact caagtattaa gagtacctta tacatataat attaaaaata
     2941 ctactaaaca agtaaaaata atacaccaag acaaaaatat atatagatat gacatagaaa
     3001 agttagctaa aaaatattgc aaagatgtaa aaacagtagg taatactaat acaaaatata
     3061 tattagatag taagctacca aattgtatag tagatatttt aaaaaatggt agtaaagatg
     3121 gacataaaaa cctagatttg caaaaaatag ttgtgacttt aagattgagg aataaaagtt
     3181 taagtcaagt aatatccgtt gctagagaat ggaactatat atcacaaaat agtctttcaa
     3241 atagtgagct agaatatcaa gtcaagtata tgtatgagaa acttaaaacg gttaattttg
     3301 gttgtactgg ttgtgagttt aatagtgatt gttggaataa aatagaatca gattttatat
     3361 atagtgatga agatactttg ttcaatatgc cacataagca ctcaaaggat ttgaaatata
     3421 agaataggaa aggggttaaa ataatgactg gtaatcaatt gtttatctat aatgtgttac
     3481 ttaacaataa agatagagaa ttaaacatag acgatataat ggagctgata acctataaac
     3541 gtaagaagaa agttaaaaac attgttatga gtgaaaagac attaagagaa acattaaaag
     3601 aacttcaaca taatgattat attacaaaaa caaaaggtgt tacaaagcta ggaataaaag
     3661 atacatacaa tgtaaaagaa gttagatgta atatagataa acaatatact attagttact
     3721 ttgttaccat ggcagtaatt tggggaataa tttcaactga agaattaaga ttatatactc
     3781 acatgagata taagcaagat ttattggtca aagatgataa aataaaagga aatatattaa
     3841 gaattaatca agaggaatta gcaaaagatt taggagtaac acagcaaaga atttcaaata
     3901 tgatagaatc tttattagat actaaaattt tagatgtatg ggaaactaaa ataaatgata
     3961 gaggatttat gtactataca tatagattaa acaagtagat ttttgatagg attagaattg
     4021 attttctagt cctattttta tgcaaaaaaa ctaattaata aaaatttctt ttggtaaaat
     4081 aattgtacga gaattgcaaa aaaaaaatgg catcaaagta ttgaaattaa gccgttttaa
     4141 aaatttcttt tggtaaaata attctacata tatatgtagt atatatatat atgtttttta
     4201 gagaatgtat aactagaata tagagctaga atatagagaa tgtataacta gaatatagag
     4261 ctagaatata gagaatgtat aactagaata tagagctaga atatagagaa tgtataacta
     4321 gaatatagag ctagaatata gagaatgtat aactagaata tagagctaga atatagagaa
     4381 tgtataacta gaatatagag ctagaatata gagaatgtat aactagaata tagagctaga
     4441 atcctaatta gtaggtgctt ttttaaaaca agttaaaaat caaaaatagt attagtaagc
     4501 attggaaatg ctagattcta aaatagaaaa gtaaaaaatt ggtgcactat ctaaacttat
     4561 ctatatcgct ttttccgtcg tttggttctc tagttacgat acaggggata tgcttatatt
     4621 gagttatagt actaatcagt gcttaatata gttaataaaa ttatagttac catagtttag
     4681 taactatgat gtatgttagt tagaaacttg catttcggcc ggccgaagca aacttaagag
     4741 tgtgttgata gtgcagtatc ttaaaatttt gtataatagg aattgaagtt aaattagatg
     4801 ctaaaaattt gtaattaaga aggagtgatt acatgaacaa aaatataaaa tattctcaaa
     4861 actttttaac gagtgaaaaa gtactcaacc aaataataaa acaattgaat ttaaaagaaa
     4921 ccgataccgt ttacgaaatt ggaacaggta aagggcattt aacgacgaaa ctggctaaaa
     4981 taagtaaaca ggtaacgtct attgaattag acagtcatct attcaactta tcgtcagaaa
     5041 aattaaaact gaatactcgt gtcactttaa ttcaccaaga tattctacag tttcaattcc
     5101 ctaacaaaca gaggtataaa attgttggga gtattcctta ccatttaagc acacaaatta
     5161 ttaaaaaagt ggtttttgaa agccatgcgt ctgacatcta tctgattgtt gaagaaggat
     5221 tctacaagcg taccttggat attcaccgaa cactagggtt gctcttgcac actcaagtct
     5281 cgattcagca attgcttaag ctgccagcgg aatgctttca tcctaaacca aaagtaaaca
     5341 gtgtcttaat aaaacttacc cgccatacca cagatgttcc agataaatat tggaagctat
     5401 atacgtactt tgtttcaaaa tgggtcaatc gagaatatcg tcaactgttt actaaaaatc
     5461 agtttcatca agcaatgaaa cacgccaaag taaacaattt aagtaccgtt acttatgagc
     5521 aagtattgtc tatttttaat agttatctat tatttaacgg gaggaaataa ttctatgagt
     5581 cgcttttgta aatttggaaa gttacacgtt actaaaggga atgtgtttaa actccttttt
     5641 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5701 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5761 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5821 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5881 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5941 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6001 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6061 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6121 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6181 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6241 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6301 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6361 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6421 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6481 gaggaagcgg aagagcgccc aatacgcagg gccc
//
