LOCUS       sequence-376569-        7934 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For red light-repressed bacterial expression of gene of interest.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7934)
  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 7934)
  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-MCS
FEATURES             Location/Qualifiers
     source          1..7934
                     /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."
     CDS             5233..5424
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(5418..5440)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5526..5665
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5581..5600
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5680..5697)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5851..6439)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5931..5950)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(6550..6569)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             6561..7376
                     /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             6561..7376
                     /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     complement(6634..6653)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(7468..7923)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7605..7626)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7817..7836
                     /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 cagtgaccaa acaggaaaaa accgccctta acatggcccg ctttatcaga
     5281 agccagacat taacgcttct ggagaaactc aacgagctgg acgcggatga acaggcagac
     5341 atctgtgaat cgcttcacga ccacgctgat gagctttacc gcagctgcct cgcgcgtttc
     5401 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     5461 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     5521 cggggcgcag ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg
     5581 cggcatcaga gcagattgta ctgagagtgc accattgcgg tgtgaaatac cgcacagatg
     5641 cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5701 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5761 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5821 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5881 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5941 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6001 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6061 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6121 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6181 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6241 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     6301 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6361 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6421 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6481 gaacgaaaac tcacgttaag ggattttggt catgaacaat aaaactgtct gcttacataa
     6541 acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcttgc tctaggccgc
     6601 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     6661 ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca gagttgtttc
     6721 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     6781 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     6841 catggttact caccactgcg atccccggga aaacagcatt ccaggtatta gaagaatatc
     6901 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     6961 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     7021 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     7081 ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg
     7141 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     7201 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     7261 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     7321 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaagaat
     7381 taattcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7441 cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg
     7501 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     7561 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     7621 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     7681 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     7741 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     7801 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     7861 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     7921 cgtcccattc gcca
//
