LOCUS       sequence-373242-        7460 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For red light-repressed bacterial expression of gene of interest 
            (StrR).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7460)
  AUTHORS   Multamaki E, Garcia de Fuentes A, Sieryi O, Bykov A, Gerken U, 
            Ranzani AT, Kohler J, Meglinski I, Moglich A, Takala H
  TITLE     Optogenetic Control of Bacterial Expression by Red Light.
  JOURNAL   ACS Synth Biol. 2022 Aug 23. doi: 10.1021/acssynbio.2c00259.
  PUBMED    35998606
REFERENCE   2  (bases 1 to 7460)
  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: "ACS Synth 
            Biol. 2022 Aug 23. doi: 10.1021/acssynbio.2c00259."
COMMENT     Sequence Label: pREDusk-StrR-MCS
FEATURES             Location/Qualifiers
     source          1..7460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(205..237)
                     /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             complement(241..258)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(268..285)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             304..326
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(655..674)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        880..957
                     /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."
     rep_origin      5233..5971
                     /direction=RIGHT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     promoter        6019..6110
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6111..6902
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      complement(6994..7449)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7131..7152)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7343..7362
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tccgacccat ttgctgtcca ccagtcatgc tagccatatg
      241 gctgccgcgc ggcaccaggc cgctgctgtg atgatgatga tgatggctgc tgcccatggt
      301 atatctcctt cttaaagtta aacaaaatta tttctagact ccgttgtgat gacgcattgg
      361 tacgcggtat cgggaggttc gaaaatttcg agcgatatct taaggggggt gccttacgta
      421 gaaccccgta ggtcatgccc gaggccggtc ctggatggcg cggcggatac gcttgagcag
      481 gttttcgtcg agaagcggct tcaaaaccac gtcttttacg ccggcctcgg cggcccgggt
      541 cgagatgttt tcgtccggat agccggtgat caggatcacg ggcgtagatc tcgatcctct
      601 acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata
      661 tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg agcgcttgtt
      721 tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgcc atctccttgc
      781 atgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg ggctgcttcc
      841 taatgcagga gtcgcataag ggagagcgtc gagatcccgg acaccatcga atggtgcaaa
      901 acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattgagggt ggtgaatgtg
      961 ctccgactca tgatcatgaa gcttttacaa agggagaccc tggacctgca ccgcgaggtc
     1021 gaggccctga tgcccgtgat gagtgagacg ctgacccggc ggggctatct ggaggtgctg
     1081 cggcgcctgc aagcggtggt cgcgccgctg gaagcacaac tcgacgccct gccgctgccg
     1141 gacgagctgg cgtggcccga gcgccgcaag gcccatctgc tcgctctgga ccttgccgta
     1201 atgggggaag ggccgtcaaa tcagcaggcg ccactgtctg tgcccgagct gaccactctg
     1261 gcgcaggcct tcggagcgct ttacgtgctt gaaggcgcga cgctaggcgg acagctcatc
     1321 agccgccatc tcaggcgcac gctggggcta agccctgagc agggcagcgc ctatttctcc
     1381 ggctatggcc cccagaccgg cccgcgctgg cgctcgttcg gggaagtgct ggaagccagc
     1441 gtgccggcag aagacgcggc agaggtcgtg gcgggagcgc ggcagacctt cggggccttc
     1501 cgccgcgcct tgcaggggct gtcggaagct gaagcggtcc aagtgcctga ggtcgcccac
     1561 gcatgagccg ggacccgttg cccttttttc caccgcttta ccttggtggc ccggaaatta
     1621 ccaccgagaa ctgcgagcgc gagccgattc atattcccgg cagcatccag ccgcacggcg
     1681 ccctgctcac tgccgacggg cacagcggcg aggtgctcca gatgagcctc aacgcggcca
     1741 cttttctggg acaggaaccc acagtgctgc gcggacagac cctcgccgca ctgctgcccg
     1801 agcagtggcc cgcgctgcaa gcggccctgc cccccggctg ccccgacgcc ctgcaatacc
     1861 gcgcaacgct ggactggcct gccgccgggc acctttcgct gacggtgcac cgggtcggcg
     1921 agttgctgat tctggaattc gagccgacgg aggcctggga cagcaccggg ccgcacgcgc
     1981 tgcgcaacgc gatgttcgcg ctcgaaagtg cccccaacct gcgggcgctg gccgaggtgg
     2041 cgacccagac ggtccgcgag ctgacgggct ttgaccgggt gatgctctac aaatttgccc
     2101 ccgacgccac cggcgaagtg attgccgagg cccgccgtga ggggctgcac gcctttctgg
     2161 gccaccgttt tcccgcgtcg gacattccgg cgcaggcccg cgcgctctac acccggcacc
     2221 tgctgcgcct gaccgccgac acccgcgccg ccgccgtgcc gctcgatccc gtcctcaacc
     2281 cgcagacgaa tgcgcccacc ccgctgggcg gcgccgtgct gcgcgccacc tcgcccatgc
     2341 acatgcagta cctgcggaac atgggcgtcg ggtcgagcct gtcggtgtcg gtggtggtcg
     2401 gcggccagct ctggggcctg atcgcctgcc accaccagac gccctacgtg ttgccgcccg
     2461 acctgcgaac cacgctcgaa tacctgggcc gcttgctgag cctgcaagtt caggtcaagg
     2521 aagcggcgga cgtggcggcc tttcgccaga gcctgcggga gcaccacgcg cgggtggccc
     2581 tcgcggcggc gcactcgctc tcgccgcacg acaccctcag tgacccggcg cttgacctgc
     2641 tgggcctgat gcgggccggg ggcctgattc tgcgtttcga gggccgctgg cagacgttgg
     2701 gtgaagtgcc gcctgccccg gcggtggacg cgctgctggc gtggctcgaa acccagccgg
     2761 gcgccctggt ccagaccgac gcgctaggcc aactgtggcc cgccggcgcc gatctcgccc
     2821 ccagcgcagc gggcctgctc gccatcagcg tgggcgaggg ctggtcggag tgcctcgtct
     2881 ggctgcggcc cgaactgcgg ctggaggtcg cctggggcgg ggccactcct gaccaggcga
     2941 aagacgacct cgggccgcgc cactcattcg acacctacct cgaagaaaaa cgcggctacg
     3001 ccgagccctg gcatcccggc gaaatcgagg aggcgcagga tctacgtgac acattgaccg
     3061 gggcgctggg cgagcgcctg ctccaggaac tgcaatccga gctcgtccac gtctccaggc
     3121 tgagcgccat gggcgaaatg gcgtccgcgc tcgcgcacga gctcaaccag ccgctggcgg
     3181 cgatcagcaa ctacatgaag ggctcgcggc ggctgcttgc cggcagcagt gatccgaaca
     3241 caccgaaggt cgaaagcgcc ctggaccgcg ccgccgagca ggcgctgcgc gccggccaga
     3301 tcatccggcg cctgcgcgac ttcgttgccc gcggcgaatc ggagaagcgg gtcgagagtc
     3361 tctccaagct gatcgaggag gccggcgcgc tcgggcttgc cggcgcgcgc gagcagaacg
     3421 tgcagctccg cttcagtctc gatccgggcg ccgatctcgt tctcgccgac cgggtgcaga
     3481 tccagcaggt cctggtcaac ctgttccgca acgcgctgga agcgatggct cagtcgcagc
     3541 gacgcgagct cgtcgtcacc aacacccccg ccgccgacga catgatcgag gtcgaagtgt
     3601 ccgacaccgg cagcggtttc caggacgacg tcattccgaa cctgtttcag actttcttca
     3661 ccaccaagga caccggcatg ggcgtgggac tgtccatcag ccgctcgatc atcgaagctc
     3721 acggcgggcg catgtgggcc gagagcaacg catcgggcgg ggcgaccttc cgcttcaccc
     3781 tcccggcagc cgacgagatg ataggaggtc tagcatgacg accaagggac atatctacgt
     3841 catcgacgac gacgcggcga tgcgggattc gctgaatttc ctgctggatt ctgccggctt
     3901 cggcgtcacg ctgtttgacg acgcgcaagc ctttctcgac gccctgccgg gtctctcctt
     3961 cggctgcgtc gtctccgacg tgcgcatgcc gggccttgac ggcatcgagc tgttgaagcg
     4021 gatgaaggcg cagcaaagcc cctttccgat cctcatcatg accggtcacg gcgacgtgcc
     4081 gctcgcggtc gaggcgatga agttaggggc ggtcgacttt ctggaaaagc ctttcgagga
     4141 cgaccgcctc accgccatga tcgaatcggc gatccgccag gccgagccgg ccgccaagag
     4201 cgaggccgtc gcgcaggata tcgccgcccg cgtcgcctcg ttgagcccca gggagcgcca
     4261 ggtcatggaa gggctgatcg ccggcctttc caacaagctg atcgcccgcg agtacgacat
     4321 cagcccgcgc accatcgagg tgtatcgggc caacgtcatg accaagatgc aggccaacag
     4381 cctttcggag ctggttcgcc tcgcgatgcg cgccggcatg ctcaacgatt gacaattgat
     4441 gtaagttagc tcactcatta ggcaccggga tctcgaccga tgcccttgag agccttcaac
     4501 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc
     4561 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat tttcggcgag
     4621 gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg
     4681 cacgccctcg ctcaagcctt cgtcactggt cccgccacca aacgtttcgg cgagaagcag
     4741 gccattatcg ccggcatggc ggccccacgg gtgcgcatga tcgtgctcct gtcgttgagg
     4801 acccggctag gctggcgggg ttgccttact ggttagcaga atgaatcacc gatacgcgag
     4861 cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc
     4921 ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt cagcgccctg caccattatg
     4981 ttccggatct gcatcgcagg atgctgctgg ctaccctgtg gaacacctac atctgtatta
     5041 acgaagcgct ggcattgacc ctgagtgatt tttctctggt cccgccgcat ccataccgcc
     5101 agttgtttac cctcacaacg ttccagtaac cgggcatgtt catcatcagt aacccgtatc
     5161 gtgagcatcc tctctcgttt catcggtatc attaccccca tgaacagaaa tcccccttac
     5221 acggaggcat cagatcaaag gatcttcttg agatcctttt tttctgcgcg taatcttttg
     5281 ccctgtaaac gaaaaaacca cctggggagg tggtttgatc gaaggttaag tcagttgggg
     5341 aactgcttaa ccgtggtaac tggctttcgc agagcacagc aaccaaatct gtccttccag
     5401 tgtagccgga ctttggcgca cacttcaaga gcaaccgcgt gtttagctaa acaaatcctc
     5461 tgcgaactcc cagttaccaa tggctgctgc cagtggcgtt ttaccgtgct tttccgggtt
     5521 ggactcaagt gaacagttac cggataaggc gcagcagtcg ggctgaacgg ggagttcttg
     5581 cttacagccc agcttggagc gaacgaccta caccgagccg agataccagt gtgtgagcta
     5641 tgagaaagcg ccacacttcc cgtaagggag aaaggcggaa caggtatccg gtaaacggca
     5701 gggtcggaac aggagagcgc aagagggagc gacccgccgg aaacggtggg gatctttaag
     5761 tcctgtcggg tttcgcccgt actgtcagat tcatggttga gcctcacggc tcccacagat
     5821 gcaccggaaa agcgtctgtt tatgtgaact ctggcaggag ggcggagcct atggaaaaac
     5881 gccaccggcg cggccctgct gttttgcctc acatgttagt cccctgctta tccacggaat
     5941 ctgtgggtaa ctttgtatgt gtccgcagcg cccgccgcag tctcacgccc ggagcgtagc
     6001 gaccgagtga gctagctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6061 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagggaag
     6121 cggtgatcgc cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc
     6181 tcgaaccgac gttgctggcc gtacatttgt acggctccgc agtggatggc ggcctgaagc
     6241 cacacagtga tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc
     6301 gagctttgat caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc
     6361 gcgctgtaga agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta
     6421 agcgcgaact gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc
     6481 cagccacgat cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg
     6541 ccttggtagg tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg
     6601 aggcgctaaa tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc
     6661 gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc
     6721 cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca
     6781 tacttgaagc tagacaggct tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag
     6841 atcagttgga agaatttgtc cactacgtga aaggcgagat caccaaggta gtcggcaaat
     6901 aagaattaat tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6961 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     7021 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     7081 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     7141 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     7201 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     7261 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     7321 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     7381 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     7441 agggcgcgtc ccattcgcca
//
