LOCUS       sequence-59070-e        6220 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone for blue-light-regulated expression of a gene of 
            interest. Expression is repressed under blue illumination..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6220)
  AUTHORS   Ohlendorf R, Vidavski RR, Eldar A, Moffat K, Moglich A
  TITLE     From dusk till dawn: one-plasmid systems for light-regulated gene 
            expression.
  JOURNAL   J Mol Biol. 2012 Mar 2;416(4):534-42. doi: 
            10.1016/j.jmb.2012.01.001. Epub 2012 Jan 8.
  PUBMED    22245580
REFERENCE   2  (bases 1 to 6220)
  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: 
            "10.1016/j.jmb.2012.01.001"; journalName: "J Mol Biol"; date: 
            "2012-03-2- 2"; volume: "416"; issue: "4"; pages: "534-42"
COMMENT     Sequence Label: pDusk
FEATURES             Location/Qualifiers
     source          1..6220
                     /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             3515..3706
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3700..3722)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3808..3950
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3863..3882
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3965..3982)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4136..4724)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4216..4235)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4835..4854)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4846..5661
                     /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             4846..5661
                     /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(4919..4938)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(5754..6209)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5891..5912)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6103..6122
                     /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 gctagttttc aatcatttgg gataccagga cagctggaag tcatcaaaaa agcacttgat
     1021 cacgtgcgag tcggtgtggt aattacagat cccgcacttg aagataatcc tattgtctac
     1081 gtaaatcaag gctttgttca aatgaccggc tacgagaccg aggaaatttt aggaaagaac
     1141 tgtcgcttct tacaggggaa acacacagat cctgcagaag tggacaacat cagaaccgct
     1201 ttacaaaata aagaaccggt caccgttcag atccaaaact acaaaaaaga cggaacgatg
     1261 ttctggaatg aattaaatat tgatccaatg gaaatagagg ataaaacgta ttttgtcggt
     1321 attcagaatg atatcaccga gcaccagcag acccaggcgc gcctccagga actgcaatcc
     1381 gagctcgtcc acgtctccag gctgagcgcc atgggcgaaa tggcgtccgc gctcgcgcac
     1441 gagctcaacc agccgctggc ggcgatcagc aactacatga agggctcgcg gcggctgctt
     1501 gccggcagca gtgatccgaa cacaccgaag gtcgaaagcg ccctggaccg cgccgccgag
     1561 caggcgctgc gcgccggcca gatcatccgg cgcctgcgcg acttcgttgc ccgcggcgaa
     1621 tcggagaagc gggtcgagag tctctccaag ctgatcgagg aggccggcgc gctcgggctt
     1681 gccggcgcgc gcgagcagaa cgtgcagctc cgcttcagtc tcgatccggg cgccgatctc
     1741 gttctcgccg accgggtgca gatccagcag gtcctggtca acctgttccg caacgcgctg
     1801 gaagcgatgg ctcagtcgca gcgacgcgag ctcgtcgtca ccaacacccc cgccgccgac
     1861 gacatgatcg aggtcgaagt gtccgacacc ggcagcggtt tccaggacga cgtcattccg
     1921 aacctgtttc agactttctt caccaccaag gacaccggca tgggcgtggg actgtccatc
     1981 agccgctcga tcatcgaagc tcacggcggg cgcatgtggg ccgagagcaa cgcatcgggc
     2041 ggggcgacct tccgcttcac cctcccggca gccgacgaga tgataggagg tctagcatga
     2101 cgaccaaggg acatatctac gtcatcgacg acgacgcggc gatgcgggat tcgctgaatt
     2161 tcctgctgga ttctgccggc ttcggcgtca cgctgtttga cgacgcgcaa gcctttctcg
     2221 acgccctgcc gggtctctcc ttcggctgcg tcgtctccga cgtgcgcatg ccgggccttg
     2281 acggcatcga gctgttgaag cggatgaagg cgcagcaaag cccctttccg atcctcatca
     2341 tgaccggtca cggcgacgtg ccgctcgcgg tcgaggcgat gaagttaggg gcggtggact
     2401 ttctggaaaa gcctttcgag gacgaccgcc tcaccgccat gatcgaatcg gcgatccgcc
     2461 aggccgagcc ggccgccaag agcgaggccg tcgcgcagga tatcgccgcc cgcgtcgcct
     2521 cgttgagccc cagggagcgc caggtcatgg aagggctgat cgccggcctt tccaacaagc
     2581 tgatcgcccg cgagtacgac atcagcccgc gcaccatcga ggtgtatcgg gccaacgtca
     2641 tgaccaagat gcaggccaac agcctttcgg agctggttcg cctcgcgatg cgcgccggca
     2701 tgctcaacga ttgacaattg atgtaagtta gctcactcat taggcaccgg gatctcgacc
     2761 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     2821 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     2881 gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga tcggcctgtc
     2941 gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac
     3001 caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccccac gggtgcgcat
     3061 gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca
     3121 gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac
     3181 ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa
     3241 gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg
     3301 tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg
     3361 gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg
     3421 ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc
     3481 catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct
     3541 taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct
     3601 ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta
     3661 ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     3721 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     3781 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
     3841 agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatata
     3901 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg
     3961 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4021 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4081 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4141 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4201 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4261 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4321 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4381 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4441 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4501 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4561 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4621 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4681 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4741 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4801 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
     4861 cgggaaacgt cttgctctag gccgcgatta aattccaaca tggatgctga tttatatggg
     4921 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg
     4981 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     5041 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     5101 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cgggaaaaca
     5161 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     5221 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     5281 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     5341 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
     5401 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     5461 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     5521 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     5581 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     5641 atgctcgatg agtttttcta agaattaatt catgagcgga tacatatttg aatgtattta
     5701 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgaaattgt
     5761 aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     5821 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     5881 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     5941 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     6001 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     6061 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     6121 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     6181 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
//
