LOCUS       sequence-373257-        8926 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For red light-induced bacterial expression of gene of interest.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8926)
  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 8926)
  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: pREDawn-MCS
FEATURES             Location/Qualifiers
     source          1..8926
                     /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(240..257)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(267..284)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             303..325
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      complement(401..428)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(444..515)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(563..595)
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag (LVA)
                     /note="mutant LVA variant that confers accelerated 
                     degradation under some conditions (Andersen et al., 1998)"
                     /translation="AANDENYALVA"
     CDS             complement(596..1306)
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     RBS             1313..1324
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(1647..1666)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1872..1949
                     /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             6225..6416
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6410..6432)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6518..6657
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6573..6592
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6672..6689)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6843..7431)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6923..6942)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(7542..7561)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             7553..8368
                     /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             7553..8368
                     /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(7626..7645)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(8460..8915)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8597..8618)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8809..8828
                     /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 tccgaccatt tgctgtccac cagtcatgct agccatatgg
      241 ctgccgcgcg gcaccaggcc gctgctgtga tgatgatgat gatggctgct gcccatggta
      301 tatctccttc ttaaagttaa acaaaattat ttctagagca accattatca ccgccagagg
      361 taaaatagtc aacacgcacg gtgttactct agtatataaa cgcagaaagg cccacccgaa
      421 ggtgagccag tgtgactcta gtagagagcg ttcaccgaca aacaacagat aaaacgaaag
      481 gcccagtctt tcgactgagc ctttcgtttt atttgatgcc tggctctagt agcgatctac
      541 actagcacta tcagcgttat taagctacta aagcgtagtt ttcgtcgttt gcagcgccaa
      601 acgtctcttc aggccactga ctagcgataa ctttccccac aacggaacaa ctctcattgc
      661 atgggatcat tgggtactgt gggtttagtg gttgtaaaaa cacctgaccg ctatccctga
      721 tcagtttctt gaaggtaaac tcatcacccc caagtctggc tatgcagaaa tcacctggct
      781 caacagcctg ctcagggtca acgagaatta acattccgtc aggaaagctt ggcttggagc
      841 ctgttggtgc ggtcatggaa ttaccttcaa cctcaagcca gaatgcagaa tcactggctt
      901 ttttggttgt gcttacccat ctctccgcat cacctttggt aaaggttcta agctcaggtg
      961 agaacatccc tgcctgaaca tgagaaaaaa cagggtactc atactcactt ctaagtgacg
     1021 gctgcatact aaccgcttca tacatctcgt agatttctct ggcgattgaa gggctaaatt
     1081 cttcaacgct aactttgaga atttttgcaa gcaatgcggc gttataagca tttaatgcat
     1141 tgatgccatt aaataaagca ccaacgcctg actgccccat ccccatcttg tctgcgacag
     1201 attcctggga taagccaagt tcatttttct ttttttcata aattgcttta aggcgacgtg
     1261 cgtcctcaag ctgctcttgt gttaatggtt tcttttttgt gctcatctag tatttctcct
     1321 cttttctaga ctccgttgtg atgacgcatt ggtacgcggt atcgggaggt tcgaaaattt
     1381 cgagcgatat cttaaggggg gtgccttacg tagaaccccg taggtcatgc ccgaggccgg
     1441 tcctggatgg cgcggcggat acgcttgagc aggttttcgt cgagaagcgg cttcaaaacc
     1501 acgtctttta cgccggcctc ggcggcccgg gtcgagatgt tttcgtccgg atagccggtg
     1561 atcaggatca cgggcgtaga tctcgatcct ctacgccgga cgcatcgtgg ccggcatcac
     1621 cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg
     1681 ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt
     1741 ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct
     1801 caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg
     1861 tcgagatccc ggacaccatc gaatggtgca aaacctttcg cggtatggca tgatagcgcc
     1921 cggaagagag tcaattgagg gtggtgaatg tgctccgact catgatcatg aagcttttac
     1981 aaagggagac cctggacctg caccgcgagg tcgaggccct gatgcccgtg atgagtgaga
     2041 cgctgacccg gcggggctat ctggaggtgc tgcggcgcct gcaagcggtg gtcgcgccgc
     2101 tggaagcaca actcgacgcc ctgccgctgc cggacgagct ggcgtggccc gagcgccgca
     2161 aggcccatct gctcgctctg gaccttgccg taatggggga agggccgtca aatcagcagg
     2221 cgccactgtc tgtgcccgag ctgaccactc tggcgcaggc cttcggagcg ctttacgtgc
     2281 ttgaaggcgc gacgctaggc ggacagctca tcagccgcca tctcaggcgc acgctggggc
     2341 taagccctga gcagggcagc gcctatttct ccggctatgg cccccagacc ggcccgcgct
     2401 ggcgctcgtt cggggaagtg ctggaagcca gcgtgccggc agaagacgcg gcagaggtcg
     2461 tggcgggagc gcggcagacc ttcggggcct tccgccgcgc cttgcagggg ctgtcggaag
     2521 ctgaagcggt ccaagtgcct gaggtcgccc acgcatgagc cgggacccgt tgcccttttt
     2581 tccaccgctt taccttggtg gcccggaaat taccaccgag aactgcgagc gcgagccgat
     2641 tcatattccc ggcagcatcc agccgcacgg cgccctgctc actgccgacg ggcacagcgg
     2701 cgaggtgctc cagatgagcc tcaacgcggc cacttttctg ggacaggaac ccacagtgct
     2761 gcgcggacag accctcgccg cactgctgcc cgagcagtgg cccgcgctgc aagcggccct
     2821 gccccccggc tgccccgacg ccctgcaata ccgcgcaacg ctggactggc ctgccgccgg
     2881 gcacctttcg ctgacggtgc accgggtcgg cgagttgctg attctggaat tcgagccgac
     2941 ggaggcctgg gacagcaccg ggccgcacgc gctgcgcaac gcgatgttcg cgctcgaaag
     3001 tgcccccaac ctgcgggcgc tggccgaggt ggcgacccag acggtccgcg agctgacggg
     3061 ctttgaccgg gtgatgctct acaaatttgc ccccgacgcc accggcgaag tgattgccga
     3121 ggcccgccgt gaggggctgc acgcctttct gggccaccgt tttcccgcgt cggacattcc
     3181 ggcgcaggcc cgcgcgctct acacccggca cctgctgcgc ctgaccgccg acacccgcgc
     3241 cgccgccgtg ccgctcgatc ccgtcctcaa cccgcagacg aatgcgccca ccccgctggg
     3301 cggcgccgtg ctgcgcgcca cctcgcccat gcacatgcag tacctgcgga acatgggcgt
     3361 cgggtcgagc ctgtcggtgt cggtggtggt cggcggccag ctctggggcc tgatcgcctg
     3421 ccaccaccag acgccctacg tgttgccgcc cgacctgcga accacgctcg aatacctggg
     3481 ccgcttgctg agcctgcaag ttcaggtcaa ggaagcggcg gacgtggcgg cctttcgcca
     3541 gagcctgcgg gagcaccacg cgcgggtggc cctcgcggcg gcgcactcgc tctcgccgca
     3601 cgacaccctc agtgacccgg cgcttgacct gctgggcctg atgcgggccg ggggcctgat
     3661 tctgcgtttc gagggccgct ggcagacgtt gggtgaagtg ccgcctgccc cggcggtgga
     3721 cgcgctgctg gcgtggctcg aaacccagcc gggcgccctg gtccagaccg acgcgctagg
     3781 ccaactgtgg cccgccggcg ccgatctcgc ccccagcgca gcgggcctgc tcgccatcag
     3841 cgtgggcgag ggctggtcgg agtgcctcgt ctggctgcgg cccgaactgc ggctggaggt
     3901 cgcctggggc ggggccactc ctgaccaggc gaaagacgac ctcgggccgc gccactcatt
     3961 cgacacctac ctcgaagaaa aacgcggcta cgccgagccc tggcatcccg gcgaaatcga
     4021 ggaggcgcag gatctacgtg acacattgac cggggcgctg ggcgagcgcc tgctccagga
     4081 actgcaatcc gagctcgtcc acgtctccag gctgagcgcc atgggcgaaa tggcgtccgc
     4141 gctcgcgcac gagctcaacc agccgctggc ggcgatcagc aactacatga agggctcgcg
     4201 gcggctgctt gccggcagca gtgatccgaa cacaccgaag gtcgaaagcg ccctggaccg
     4261 cgccgccgag caggcgctgc gcgccggcca gatcatccgg cgcctgcgcg acttcgttgc
     4321 ccgcggcgaa tcggagaagc gggtcgagag tctctccaag ctgatcgagg aggccggcgc
     4381 gctcgggctt gccggcgcgc gcgagcagaa cgtgcagctc cgcttcagtc tcgatccggg
     4441 cgccgatctc gttctcgccg accgggtgca gatccagcag gtcctggtca acctgttccg
     4501 caacgcgctg gaagcgatgg ctcagtcgca gcgacgcgag ctcgtcgtca ccaacacccc
     4561 cgccgccgac gacatgatcg aggtcgaagt gtccgacacc ggcagcggtt tccaggacga
     4621 cgtcattccg aacctgtttc agactttctt caccaccaag gacaccggca tgggcgtggg
     4681 actgtccatc agccgctcga tcatcgaagc tcacggcggg cgcatgtggg ccgagagcaa
     4741 cgcatcgggc ggggcgacct tccgcttcac cctcccggca gccgacgaga tgataggagg
     4801 tctagcatga cgaccaaggg acatatctac gtcatcgacg acgacgcggc gatgcgggat
     4861 tcgctgaatt tcctgctgga ttctgccggc ttcggcgtca cgctgtttga cgacgcgcaa
     4921 gcctttctcg acgccctgcc gggtctctcc ttcggctgcg tcgtctccga cgtgcgcatg
     4981 ccgggccttg acggcatcga gctgttgaag cggatgaagg cgcagcaaag cccctttccg
     5041 atcctcatca tgaccggtca cggcgacgtg ccgctcgcgg tcgaggcgat gaagttaggg
     5101 gcggtcgact ttctggaaaa gcctttcgag gacgaccgcc tcaccgccat gatcgaatcg
     5161 gcgatccgcc aggccgagcc ggccgccaag agcgaggccg tcgcgcagga tatcgccgcc
     5221 cgcgtcgcct cgttgagccc cagggagcgc caggtcatgg aagggctgat cgccggcctt
     5281 tccaacaagc tgatcgcccg cgagtacgac atcagcccgc gcaccatcga ggtgtatcgg
     5341 gccaacgtca tgaccaagat gcaggccaac agcctttcgg agctggttcg cctcgcgatg
     5401 cgcgccggca tgctcaacga ttgacaattg atgtaagtta gctcactcat taggcaccgg
     5461 gatctcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
     5521 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
     5581 tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga
     5641 tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg
     5701 gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccccac
     5761 gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta
     5821 ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa
     5881 cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg
     5941 aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct
     6001 ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga
     6061 tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta
     6121 accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta
     6181 tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa
     6241 aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac
     6301 tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg
     6361 atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     6421 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     6481 gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta
     6541 gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt
     6601 gcaccattgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     6661 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     6721 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     6781 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     6841 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6901 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6961 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     7021 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     7081 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     7141 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     7201 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     7261 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     7321 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     7381 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     7441 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     7501 gtcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     7561 tattcaacgg gaaacgtctt gctctaggcc gcgattaaat tccaacatgg atgctgattt
     7621 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt
     7681 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     7741 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     7801 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     7861 gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     7921 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     7981 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     8041 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     8101 taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     8161 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     8221 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     8281 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     8341 tcatttgatg ctcgatgagt ttttctaaga attaattcat gagcggatac atatttgaat
     8401 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     8461 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     8521 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     8581 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     8641 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     8701 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     8761 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     8821 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     8881 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgcca
//
