LOCUS       sequence-375364-        6299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Clostridium expression vector (pAMB1 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 6299)
  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 6299)
  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: pATmin-GG
FEATURES             Location/Qualifiers
     source          1..6299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    937..958
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(973..978,979..996,997..1003)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1011..1027
                     /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             1045..1056
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1063..1740
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1785..1856
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1872..1899
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct agtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcatttc tagaaggatg agacctcaat
      841 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      901 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      961 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     1021 ataacaattt cacacatact agagaaagag gagaaatact agatggcttc ctccgaagac
     1081 gttatcaaag agttcatgcg tttcaaagtt cgtatggaag gttccgttaa cggtcacgag
     1141 ttcgaaatcg aaggtgaagg tgaaggtcgt ccgtacgaag gtacccagac cgctaaactg
     1201 aaagttacca aaggtggtcc gctgccgttc gcttgggaca tcctgtcccc gcagttccag
     1261 tacggttcca aagcttacgt taaacacccg gctgacatcc cggactacct gaaactgtcc
     1321 ttcccggaag gtttcaaatg ggaacgtgtt atgaacttcg aagacggtgg tgttgttacc
     1381 gttacccagg actcctccct gcaagacggt gagttcatct acaaagttaa actgcgtggt
     1441 accaacttcc cgtccgacgg tccggttatg cagaaaaaaa ccatgggttg ggaagcttcc
     1501 accgaacgta tgtacccgga agacggtgct ctgaaaggtg aaatcaaaat gcgtctgaaa
     1561 ctgaaagacg gtggtcacta cgacgctgaa gttaaaacca cctacatggc taaaaaaccg
     1621 gttcagctgc cgggtgctta caaaaccgac atcaaactgg acatcacctc ccacaacgaa
     1681 gactacacca tcgttgaaca gtacgaacgt gctgaaggtc gtcactccac cggtgcttaa
     1741 taacgctgat agtgctagtg tagatcgcta ctagagccag gcatcaaata aaacgaaagg
     1801 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact
     1861 agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttataactat ggtctcccga
     1921 atcttccctc gagtatttat ttctattgag taaaataaaa aagatggtga gataatctca
     1981 ccatcttttt ttcttttaaa tccgtcgacc ttagaagcaa acttaagagt gtgttgatag
     2041 tgcagtatct taaaattttg tgtataatag gaattgaagt taaattagat gctaaaaatt
     2101 tgtaattaag aaggagggat tcgtcatgtt ggtattccaa atgcgtaatg tagataaaac
     2161 atctactgtt ttgaaacaga ctaaaaacag tgattacgca gataaataaa tacgttagat
     2221 taattcctac cagtgactaa tcttatgact ttttaaacag ataactaaaa ttacaaacaa
     2281 atcgtttaac ttctgtattt atttacagat gtaatcactt caggagtaat tacatgaaca
     2341 aaaatataaa atattctcaa aactttttaa cgagtgaaaa agtactcaac caaataataa
     2401 aacaattgaa tttaaaagaa accgataccg tttacgaaat tggaacaggt aaagggcatt
     2461 taacgacgaa actggctaaa ataagtaaac aggtaacgtc tattgaatta gacagtcatc
     2521 tattcaactt atcgtcagaa aaattaaaac tgaacattcg tgtcacttta attcaccaag
     2581 atattctaca gtttcaattc cctaacaaac agaggtataa aattgttggg agtattcctt
     2641 accatttaag cacacaaatt attaaaaaag tggtttttga aagccatgcg tctgacatct
     2701 atctgattgt tgaagaagga ttctacaagc gtaccttgga tattcaccga acactagggt
     2761 tgctcttgca cactcaagtc tcgattcagc aattgcttaa gctgccagcg gaatgctttc
     2821 atcctaaacc aaaagtaaac agtgtcttaa taaaacttac ccgccatacc acagatgttc
     2881 cagataaata ttggaagcta tatacgtact ttgtttcaaa atgggtcaat cgagaatatc
     2941 gtcaactgtt tactaaaaat cagtttcatc aagcaatgaa acacgccaaa gtaaacaatt
     3001 taagtaccat tacttatgag caagtattgt ctatttttaa tagttatcta ttatttaacg
     3061 ggaggaaata attctatgag tcgctttttt aaatttggaa agttacacgt tactaaaggg
     3121 aatggagata aattattaga tatactactg acagcttcca agaagctaaa gaggtcccta
     3181 gcgcctacgg ggaatttggg caaatcaaaa gaaaaattct cccaagaatt agaaatgagt
     3241 agatcaaatt attcacgaat agaatcagga aaatcagatc caaccataaa aacactagaa
     3301 caaattgcaa agttaactaa ctcaacgcta gtagtggatt taatcccaaa tgagccaaca
     3361 gaaccagaac cagaaacaga atcagaacaa gtaacattgg atttagaaat ggaagaagaa
     3421 aaaagcaatg acttcgtgtg aataatgcac gaaatcgttg cttatttttt ttaaaagcgg
     3481 tatactagat ataacgaaac aacgaactga atagaaacga aaaaagagcc atgacacatt
     3541 tataaaatgt ttgacgacat tttataaatg catagcccga taagattgcc aaaccaacgc
     3601 ttatcagtta gtcagatgaa ctcttccctc gtaagaagtt atttaattaa ctttgtttga
     3661 agacggtata taaccgtact atcattatat agggaaatca gagagttttc aagtatctaa
     3721 gctactgaat ttaagaattg ttaagcaatc aatcggaaat cgtttgattg ctttttttgt
     3781 attcatttat agaaggtgga gtttgtatga atcatgatga atgtaaaact tatataaaaa
     3841 atagtttatt ggagataaga aaattagcaa atatctatac actagaaacg tttaagaaag
     3901 agttagaaaa gagaaatatc tacttagaaa caaaatcaga taagtatttt tcttcggagg
     3961 gggaagatta tatatataag ttaatagaaa ataacaaaat aatttattcg attagtggaa
     4021 aaaaattgac ttataaagga aaaaaatctt tttcaaaaca tgcaatattg aaacagttga
     4081 atgaaaaagc aaaccaagtt aattaaacaa cctattttat aggatttata ggaaaggaga
     4141 acagctgaat gaatatccct tttgttgtag aaactgtgct tcatgacggc ttgttaaagt
     4201 acaaatttaa aaatagtaaa attcgctcaa tcactaccaa gccaggtaaa agcaaagggg
     4261 ctatttttgc gtatcgctca aaatcaagca tgattggcgg tcgtggtgtt gttctgactt
     4321 ccgaggaagc gattcaagaa aatcaagata catttacaca ttggacaccc aacgtttatc
     4381 gttatggaac gtatgcagac gaaaaccgtt catacacgaa aggacattct gaaaacaatt
     4441 taagacaaat caataccttc tttattgatt ttgatattca cacggcaaaa gaaactattt
     4501 cagcaagcga tattttaaca accgctattg atttaggttt tatgcctact atgattatca
     4561 aatctgataa aggttatcaa gcatattttg ttttagaaac gccagtctat gtgacttcaa
     4621 aatcagaatt taaatctgtc aaagcagcca aaataatttc gcaaaatatc cgagaatatt
     4681 ttggaaagtc tttgccagtt gatctaacgt gtaatcattt tggtattgct cgcataccaa
     4741 gaacggacaa tgtagaattt tttgatccta attaccgtta ttctttcaaa gaatggcaag
     4801 attggtcttt caaacaaaca gataataagg gctttactcg ttcaagtcta acggttttaa
     4861 gcggtacaga aggcaaaaaa caagtagatg aaccctggtt taatctctta ttgcacgaaa
     4921 cgaaattttc aggagaaaag ggtttaatag ggcgtaataa cgtcatgttt accctctctt
     4981 tagcctactt tagttcaggc tattcaatcg aaacgtgcga atataatatg tttgagttta
     5041 ataatcgatt agatcaaccc ttagaagaaa aagaagtaat caaaattgtt agaagtgcct
     5101 attcagaaaa ctatcaaggg gctaataggg aatacattac cattctttgc aaagcttggg
     5161 tatcaagtga tttaaccagt aaagatttat ttgtccgtca agggtggttt aaattcaaga
     5221 aaaaaagaag cgaacgtcaa cgtgttcatt tgtcagaatg gaaagaagat ttaatggctt
     5281 atattagcga aaaaagcgat gtatacaagc cttatttagt gacgaccaaa aaagagatta
     5341 gagaagtgct aggcattcct gaacggacat tagataaatt gctgaaggta ctgaaggcga
     5401 atcaggaaat tttctttaag attaaaccag gaagaaatgg tggcattcaa cttgctagtg
     5461 ttaaatcatt gttgctatcg atcattaaag taaaaaaaga agaaaaagaa agctatataa
     5521 aggcgctgac aaattctttt gacttagagc atacattcat tcaagagact ttaaacaagc
     5581 tagcagaacg ccctaaaacg gacacacaac tcgatttgtt tagctatgat acaggctgaa
     5641 aataaaaccc gcactatgcc attacattta tatctatgat acgtgtttgt tttttctttg
     5701 ctgtttagcg aatgattagc agaaatatac agagtaagat tttaattaat tattaggggg
     5761 agaaggagag agtagcccga aaacttttag ttggcttgga ctgaacgaag tgagggaaag
     5821 gctactaaaa cgtcgagggg cagtgagagc gaagcgaaca cttgattttt taattttcta
     5881 tcttttatag gtcattagag tatacttatt tgtcctataa actatttagc agcataatag
     5941 atttattgaa taggtcattt aagttgagca tattagagga ggaaaatctt ggagaaatat
     6001 ttgaagaacc cgattacatg gattggatta gttcttgtgg ttacgtggtt tttaactaaa
     6061 agtagtgaat ttttgatttt tggtgtgtgt gtcttgttgt tagtatttgc tagtcaaagt
     6121 gattagatac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6181 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6241 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttc
//
