LOCUS       Exported                7213 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pDawn, complete sequence.
ACCESSION   JN579121
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7213)
  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. 416 (4), 534-542 (2012)
  PUBMED    22245580
REFERENCE   2  (bases 1 to 7213)
  AUTHORS   Ohlendorf R, Moffat K, Moeglich A.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-AUG-2011) Biophysikalische Chemie, Humboldt 
            Universitaet zu Berlin, Invalidenstrasse 42, Berlin 10115, Germany
REFERENCE   3  (bases 1 to 7213)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7213
                     /organism="Expression vector pDawn"
                     /mol_type="other DNA"
                     /label=vector for blue-light-induced gene expression
                     /note="vector for blue-light-induced gene expression"
                     /db_xref="taxon:1170225"
     regulatory      complement(26..73)
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      26..73
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    complement(158..203)
                     /label=multiple cloning site
                     /note="multiple cloning site"
     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)"
     regulatory      complement(339..387)
                     /regulatory_class="promoter"
                     /note="lambda pR promoter"
     terminator      402..429
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      445..516
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(561..1307)
                     /codon_start=1
                     /product="cI-LVA"
                     /label=cI-LVA
                     /note="lambda cI-LVA"
                     /protein_id="AFH34992.1"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFGAANDENYALVA"
     CDS             complement(564..596)
                     /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(597..1307)
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=-lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     RBS             1314..1325
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     regulatory      complement(1332..1578)
                     /regulatory_class="promoter"
                     /note="FixK2 promoter"
     promoter        1873..1950
                     /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."
     misc_feature    1951..3704
                     /label=YF1/FixJ bicistronic operon
                     /note="YF1/FixJ bicistronic operon"
     CDS             1951..3093
                     /codon_start=1
                     /product="YF1"
                     /label=YF1
                     /protein_id="AFH34994.1"
                     /translation="MASFQSFGIPGQLEVIKKALDHVRVGVVITDPALEDNPIVYVNQG
                     FVQMTGYETEEILGKNCRFLQGKHTDPAEVDNIRTALQNKEPVTVQIQNYKKDGTMFWN
                     ELNIDPMEIEDKTYFVGIQNDITEHQQTQARLQELQSELVHVSRLSAMGEMASALAHEL
                     NQPLAAISNYMKGSRRLLAGSSDPNTPKVESALDRAAEQALRAGQIIRRLRDFVARGES
                     EKRVESLSKLIEEAGALGLAGAREQNVQLRFSLDPGADLVLADRVQIQQVLVNLFRNAL
                     EAMAQSQRRELVVTNTPAADDMIEVEVSDTGSGFQDDVIPNLFQTFFTTKDTGMGVGLS
                     ISRSIIEAHGGRMWAESNASGGATFRFTLPAADEMIGGLA"
     CDS             3090..3707
                     /codon_start=1
                     /product="FixJ"
                     /label=FixJ
                     /protein_id="AFH34995.1"
                     /translation="MTTKGHIYVIDDDAAMRDSLNFLLDSAGFGVTLFDDAQAFLDALP
                     GLSFGCVVSDVRMPGLDGIELLKRMKAQQSPFPILIMTGHGDVPLAVEAMKLGAVDFLE
                     KPFEDDRLTAMIESAIRQAEPAAKSEAVAQDIAARVASLSPRERQVMEGLIAGLSNKLI
                     AREYDISPRTIEVYRANVMTKMQANSLSELVRLAMRAGMLND"
     CDS             4508..4699
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    4801..4943
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5111..5793)
                     /direction=LEFT
                     /label=ColE1 origin
                     /note="ColE1 origin"
     rep_origin      complement(5129..5717)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             5839..6654
                     /codon_start=1
                     /product="KanR"
                     /label=KanR
                     /note="KanR resistance marker"
                     /protein_id="AFH34993.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      complement(6747..7202)
                     /direction=LEFT
                     /label=M13 origin
                     /note="M13 origin"
     rep_origin      complement(6879..7185)
                     /direction=LEFT
                     /label=F1 origin
                     /note="F1 origin"