LOCUS       sequence-422226-        7985 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for red-light-repressed bacterial gene expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7985)
  AUTHORS   Meier SSM, Multamaki E, Ranzani AT, Takala H, Moglich A
  TITLE     Leveraging the histidine kinase-phosphatase duality to sculpt 
            two-component signaling.
  JOURNAL   Nat Commun. 2024 Jun 10;15(1):4876. doi: 10.1038/s41467-024-49251-8.
  PUBMED    38858359
REFERENCE   2  (bases 1 to 7985)
  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"; journalName: "Nat 
            Commun."; date: "2024-06-10"; volume: "15(1):4876. doi"; pages: " 
            10.1038/s41467-024-49251-8"
COMMENT     Sequence Label: pDmREDusk
FEATURES             Location/Qualifiers
     source          1..7985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(81..100)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     291..312
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(541..1356)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(541..1356)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1264..1283
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1348..1367
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1478..2066
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1967..1986
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2220..2237
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2252..2391
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2317..2336)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2477..2499
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2493..2684)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        complement(7011..7088)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     7294..7313
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             7642..7664
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             7683..7700
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             7710..7727
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             7731..7763
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             7811..7828
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7881..7899)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7895..7942
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      join(7979..7985,1..449)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
       61 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
      121 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
      181 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
      241 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
      301 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
      361 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
      421 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
      481 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc
      541 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
      601 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
      661 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
      721 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
      781 tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca
      841 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
      901 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
      961 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     1021 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
     1081 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     1141 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     1201 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     1261 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     1321 cctagagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
     1381 gtaagcagac agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact
     1441 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1501 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1561 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1621 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1681 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1741 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     1801 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     1861 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     1921 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     1981 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2041 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2101 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2161 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2221 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     2281 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     2341 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     2401 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     2461 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     2521 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     2581 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     2641 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     2701 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     2761 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     2821 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     2881 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     2941 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     3001 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     3061 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     3121 aacgacagga gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc
     3181 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     3241 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     3301 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca
     3361 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     3421 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatca
     3481 attgtcaatc gttgagcatg ccggcgcgca tcgcgaggcg aaccagctcc gaaaggctgt
     3541 tggcctgcat cttggtcatg acgttggccc gatacacctc gatggtgcgc gggctgatgt
     3601 cgtactcgcg ggcgatcagc ttgttggaaa ggccggcgat cagcccttcc atgacctggc
     3661 gctccctggg gctcaacgag gcgacgcggg cggcgatatc ctgcgcgacg gcctcgctct
     3721 tggcggccgg ctcggcctgg cggatcgccg attcgatcat ggcggtgagg cggtcgtcct
     3781 cgaaaggctt ttccagaaag tcgaccgccc ctaacttcat cgcctcgacc gcgagcggca
     3841 cgtcgccgtg accggtcatg atgaggatcg gaaaggggct ttgctgcgcc ttcatccgct
     3901 tcaacagctc gatgccgtca aggcccggca tgcgcacgtc ggagacgacg cagccgaagg
     3961 agagacccgg cagggcgtcg agaaaggctt gcgcgtcgtc aaacagcgtg acgccgaagc
     4021 cggcagaatc cagcaggaaa ttcagcgaat cccgcatcgc cgcgtcgtcg tcgatgacgt
     4081 agatatgtcc cttggtcgtc atgctagacc tcctatcatc tcgtcggctg ccgggagggt
     4141 gaagcggaag gtcgccccgc ccgatgcgtt gctctcggcc cacatgcgcc cgccgtgagc
     4201 ttcgatgatc gagcggctga tggacagtcc cacgcccatg ccggtgtcct tggtggtgaa
     4261 gaaagtctga aacaggttcg gaatgacgtc gtcctggaaa ccgctgccgg tgtcggacac
     4321 ttcgacctcg atcatgtcgt cggcggcggg ggtgttggtg acgacgagct cgcgtcgctg
     4381 cgactgagcc atcgcttcca gcgcgttgcg gaacaggttg accaggacct gctggatctg
     4441 cacccggtcg gcgagaacga gatcggcgcc cggatcgaga ctgaagcgga gctgcacgtt
     4501 ctgctcgcgc gcgccggcaa gcccgagcgc gccggcctcc tcgatcagct tggagagact
     4561 ctcgacccgc ttctccgatt cgccgcgggc aacgaagtcg cgcaggcgcc ggatgatctg
     4621 gccggcgcgc agcgcctgct cggcggcgcg gtccagggcg ctttcgacct tcggtgtgtt
     4681 cggatcactg ctgccggcaa gcagccgccg cgagcccttc atgtagttgc tgatcgccgc
     4741 cagcggctgg ttgagctcgt gcgcgagcgc ggacgccatt tcgcccatgg cgctcagcct
     4801 ggagacgtgg acgagctcgg attgcagttc ctggagcaga cgggcaccct gaacttccag
     4861 taatgcacca cgcagatcca gtgctgcatc cagatcgcct gcctgccacg gctgggcaac
     4921 accacgaact gcttcaacat agctgctaaa gctggcacgc ggagtcagac gtgctgtacc
     4981 atcatctgaa acctgaacgg ctttatgcgg atcaccaccc caggtaatgg ttgccggaat
     5041 ttccggacga aaccacagca gatattcatc ccaattaccg ctaatgctca cacccagaac
     5101 accgcttgca cgttccataa acagtgcacc tggcggatag gtacgtgcca gcgcatcggt
     5161 acaaacgctc ggtgtaccct gtgcacgcag ccatgcaacc agatgttcca gatcctcatc
     5221 actcggggta acacccaggg tacgtgtacc accatccata cgaactgcgg caccggctgt
     5281 atccagaaaa ccaggcagat tcagatcggc acgggtcagt gcatccagcg gtgtcagcgt
     5341 accggtaact gcggtaacca gacgtgcatg ctgggcaccc agggctgcac gacgggcatt
     5401 aactgctgca tcacgttttg cggtaatctg caggctcaga acacgtccca gaaattcaca
     5461 tgcatcacgg gttgcctgcg gaacaacatg tgcggcacca tgatgacatg caatcagacc
     5521 ccacagacga ccatcctgaa caatgctaac gctcatgctt gcaataacac ccatattacg
     5581 cagatactgc agatgaaccg gactggttgc acgtaaaacc agggcaccca tctgcagcgg
     5641 tgcattggta accggattca gtgcaggaac cagcggaacc tgaccaccac cggcatctgc
     5701 ggtaaaacgc agtaaatgct gcacatacag cgcacgtgcc tgacgcggaa tatctgattc
     5761 cggaaaacga tgacccagaa atgcatgtaa atgcggtgca cgtgcttcgg caacaacttc
     5821 accgctatca tctgctgcaa aacgatagat catcacacga tccagaccgg tcagggcacg
     5881 aacacgctgt gctgcaacat ccagcagggc atgcagatcc ggtgcatgtt ccagtgcacc
     5941 cagtgcatca cgaattgcat gatacagtgc cggagtgcct gcacgaacat caggcggttc
     6001 cagttccatc agcagcagtt cgccatcacg acgtgcggta acgtcatagg tacgaccatt
     6061 acgaaaggtc agggtcattg cgctcggacc tgcctgtgca aatgcggcac gcagtgcggc
     6121 tgcaccctga tcaccaacca gatcggccag atgctgaccc agcagcgcat caggtgcaac
     6181 accaacaaat gcttcgctat ttgcgctaac ctgaactgca cgttcatcat ctgcacgcag
     6241 aaccagcagt gcaccatgcg gctgaacggc acccggaata tgaatcggtt cacgtgcaca
     6301 attatcggta tcaacaggcg gaccacctaa atgtgccggt gtcagaaaat catcaccacc
     6361 ggtcatacgt gctgcacccg gacgcgctgc cggacgttct gtcatgcgtg ggcgacctca
     6421 ggcacttgga ccgcttcagc ttccgacagc ccctgcaagg cgcggcggaa ggccccgaag
     6481 gtctgccgcg ctcccgccac gacctctgcc gcgtcttctg ccggcacgct ggcttccagc
     6541 acttccccga acgagcgcca gcgcgggccg gtctgggggc catagccgga gaaataggcg
     6601 ctgccctgct cagggcttag ccccagcgtg cgcctgagat ggcggctgat gagctgtccg
     6661 cctagcgtcg cgccttcaag cacgtaaagc gctccgaagg cctgcgccag agtggtcagc
     6721 tcgggcacag acagtggcgc ctgctgattt gacggccctt cccccattac ggcaaggtcc
     6781 agagcgagca gatgggcctt gcggcgctcg ggccacgcca gctcgtccgg cagcggcagg
     6841 gcgtcgagtt gtgcttccag cggcgcgacc accgcttgca ggcgccgcag cacctccaga
     6901 tagccccgcc gggtcagcgt ctcactcatc acgggcatca gggcctcgac ctcgcggtgc
     6961 aggtccaggg tctccctttg taaaagcttc atgatcatga gtcggagcac attcaccacc
     7021 ctcaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcaccattcg
     7081 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     7141 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
     7201 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     7261 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     7321 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat
     7381 ctacgcccgt gatcctgatc accggctatc cggacgaaaa catctcgacc cgggccgccg
     7441 aggccggcgt aaaagacgtg gttttgaagc cgcttctcga cgaaaacctg ctcaagcgta
     7501 tccgccgcgc catccaggac cggcctcggg catgacctac ggggttctac gtaaggcacc
     7561 ccccttaaga tatcgctcga aattttcgaa cctcccgata ccgcgtacca atgcgtcatc
     7621 acaacggagt ctagaaataa ttttgtttaa ctttaagaag gagatatacc atgggcagca
     7681 gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat atggctagca
     7741 tgactggtgg acagcaaatg ggtcggatcc gaattcgagc tccgtcgaca agcttgcggc
     7801 cgcactcgag caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga
     7861 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     7921 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac
     7981 gcgcc
//
