LOCUS       sequence-352655-        7209 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone for blue-light-regulated expression of a gene of 
            interest. Expression is induced under blue illumination..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7209)
  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 7209)
  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: pDawn
FEATURES             Location/Qualifiers
     source          1..7209
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1353..1544
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1538..1560)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1646..1785
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1701..1720
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1800..1817)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1971..2559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2051..2070)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2670..2689)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2681..3496
                     /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             2681..3496
                     /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(2754..2773)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(3588..4043)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3725..3746)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3937..3956
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(4080..4127)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4123..4141
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4194..4211)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4259..4291)
                     /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(4295..4312)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(4322..4339)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             4358..4380
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      complement(4456..4483)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(4499..4570)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4618..4650)
                     /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(4651..5361)
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     RBS             5368..5379
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(5702..5721)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5927..6004
                     /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."
ORIGIN
        1 ctgctggatt ctgccggctt cggcgtcacg ctgtttgacg acgcgcaagc ctttctcgac
       61 gccctgccgg gtctctcctt cggctgcgtc gtctccgacg tgcgcatgcc gggccttgac
      121 ggcatcgagc tgttgaagcg gatgaaggcg cagcaaagcc cctttccgat cctcatcatg
      181 accggtcacg gcgacgtgcc gctcgcggtc gaggcgatga agttaggggc ggtggacttt
      241 ctggaaaagc ctttcgagga cgaccgcctc accgccatga tcgaatcggc gatccgccag
      301 gccgagccgg ccgccaagag cgaggccgtc gcgcaggata tcgccgcccg cgtcgcctcg
      361 ttgagcccca gggagcgcca ggtcatggaa gggctgatcg ccggcctttc caacaagctg
      421 atcgcccgcg agtacgacat cagcccgcgc accatcgagg tgtatcgggc caacgtcatg
      481 accaagatgc aggccaacag cctttcggag ctggttcgcc tcgcgatgcg cgccggcatg
      541 ctcaacgatt gacaattgat gtaagttagc tcactcatta ggcaccggga tctcgaccga
      601 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
      661 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
      721 tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc
      781 ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca
      841 aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccccacgg gtgcgcatga
      901 tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga
      961 atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct
     1021 gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt
     1081 cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg
     1141 gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt tttctctggt
     1201 cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac cgggcatgtt
     1261 catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc attaccccca
     1321 tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa accgccctta
     1381 acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg
     1441 acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc
     1501 gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     1561 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     1621 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
     1681 tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accattgcgg
     1741 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc
     1801 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     1861 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1921 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1981 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2041 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2101 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2161 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2221 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2281 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2341 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2401 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2461 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2521 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2581 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgaacaat
     2641 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     2701 aacgtcttgc tctaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     2761 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc
     2821 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2881 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2941 tatccgtact cctgatgatg catggttact caccactgcg atccccggga aaacagcatt
     3001 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     3061 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     3121 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     3181 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     3241 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     3301 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     3361 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     3421 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     3481 cgatgagttt ttctaagaat taattcatga gcggatacat atttgaatgt atttagaaaa
     3541 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt
     3601 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     3661 gaccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     3721 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     3781 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     3841 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     3901 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     3961 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     4021 taatgcgccg ctacagggcg cgtcccattc gccaatccgg atatagttcc tcctttcagc
     4081 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctagtta ttgctcagcg
     4141 gtggcagcag ccaactcagc ttcctttcgg gctttgttag cagccggatc tcagtggtgg
     4201 tggtggtggt gctcgagtgc ggccgcaagc ttgtcgacgg agctcgaatt cggatccgac
     4261 ccatttgctg tccaccagtc atgctagcca tatggctgcc gcgcggcacc aggccgctgc
     4321 tgtgatgatg atgatgatgg ctgctgccca tggtatatct ccttcttaaa gttaaacaaa
     4381 attatttcta gagcaaccat tatcaccgcc agaggtaaaa tagtcaacac gcacggtgtt
     4441 actctagtat ataaacgcag aaaggcccac ccgaaggtga gccagtgtga ctctagtaga
     4501 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtctttcgac tgagcctttc
     4561 gttttatttg atgcctggct ctagtagcga tctacactag cactatcagc gttattaagc
     4621 tactaaagcg tagttttcgt cgtttgcagc gccaaacgtc tcttcaggcc actgactagc
     4681 gataactttc cccacaacgg aacaactctc attgcatggg atcattgggt actgtgggtt
     4741 tagtggttgt aaaaacacct gaccgctatc cctgatcagt ttcttgaagg taaactcatc
     4801 acccccaagt ctggctatgc agaaatcacc tggctcaaca gcctgctcag ggtcaacgag
     4861 aattaacatt ccgtcaggaa agctcggctt ggagcctgtt ggtgcggtca tggaattacc
     4921 ttcaacctca agccagaatg cagaatcact ggcttttttg gttgtgctta cccatctctc
     4981 cgcatcacct ttggtaaagg ttctaagctc aggtgagaac atccctgcct gaacatgaga
     5041 aaaaacaggg tactcatact cacttctaag tgacggctgc atactaaccg cttcatacat
     5101 ctcgtagatt tctctggcga ttgaagggct aaattcttca acgctaactt tgagaatttt
     5161 tgcaagcaat gcggcgttat aagcatttaa tgcattgatg ccattaaata aagcaccaac
     5221 gcctgactgc cccatcccca tcttgtctgc gacagattcc tgggataagc caagttcatt
     5281 tttctttttt tcataaattg ctttaaggcg acgtgcgtcc tcaagctgct cttgtgttaa
     5341 tggtttcttt tttgtgctca tctagtattt ctcctctttt ctagactccg ttgtgatgac
     5401 gcattggtac gcggtatcgg gaggttcgaa aatttcgagc gatatcttaa ggggggtgcc
     5461 ttacgtagaa ccccgtaggt catgcccgag gccggtcctg gatggcgcgg cggatacgct
     5521 tgagcaggtt ttcgtcgaga agcggcttca aaaccacgtc ttttacgccg gcctcggcgg
     5581 cccgggtcga gatgttttcg tccggatagc cggtgatcag gatcacgggc gtagatctcg
     5641 atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc
     5701 gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc
     5761 gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc
     5821 tccttgcatg caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc
     5881 tgcttcctaa tgcaggagtc gcataaggga gagcgtcgag atcccggaca ccatcgaatg
     5941 gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat tgagggtggt
     6001 gaatgtggct agttttcaat catttgggat accaggacag ctggaagtca tcaaaaaagc
     6061 acttgatcac gtgcgagtcg gtgtggtaat tacagatccc gcacttgaag ataatcctat
     6121 tgtctacgta aatcaaggct ttgttcaaat gaccggctac gagaccgagg aaattttagg
     6181 aaagaactgt cgcttcttac aggggaaaca cacagatcct gcagaagtgg acaacatcag
     6241 aaccgcttta caaaataaag aaccggtcac cgttcagatc caaaactaca aaaaagacgg
     6301 aacgatgttc tggaatgaat taaatattga tccaatggaa atagaggata aaacgtattt
     6361 tgtcggaatt cagaatgata tcaccgagca ccagcagacc caggcgcgcc tccaggaact
     6421 gcaatccgag ctcgtccacg tctccaggct gagcgccatg ggcgaaatgg cgtccgcgct
     6481 cgcgcacgag ctcaaccagc cgctggcggc gatcagcaac tacatgaagg gctcgcggcg
     6541 gctgcttgcc ggcagcagtg atccgaacac accgaaggtc gaaagcgccc tggaccgcgc
     6601 cgccgagcag gcgctgcgcg ccggccagat catccggcgc ctgcgcgact tcgttgcccg
     6661 cggcgaatcg gagaagcggg tcgagagtct ctccaagctg atcgaggagg ccggcgcgct
     6721 cgggcttgcc ggcgcgcgcg agcagaacgt gcagctccgc ttcagtctcg atccgggcgc
     6781 cgatctcgtt ctcgccgacc gggtgcagat ccagcaggtc ctggtcaacc tgttccgcaa
     6841 cgcgctggaa gcgatggctc agtcgcagcg acgcgagctc gtcgtcacca acacccccgc
     6901 cgccgacgac atgatcgagg tcgaagtgtc cgacaccggc agcggtttcc aggacgacgt
     6961 cattccgaac ctgtttcaga ctttcttcac caccaaggac accggcatgg gcgtgggact
     7021 gtccatcagc cgctcgatca tcgaagctca cggcgggcgc atgtgggccg agagcaacgc
     7081 atcgggcggg gcgaccttcc gcttcaccct cccggcagcc gacgagatga taggaggtct
     7141 agcatgacga ccaagggaca tatctacgtc atcgacgacg acgcggcgat gcgggattcg
     7201 ctgaatttc
//
