LOCUS       V008067                 7213 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V008067
VERSION     V008067
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7213)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. Mol.
            Biol."; date: "2012"; volume: "416"; issue: "4"; pages: "534-542"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (15-AUG-2011) Biophysikalische Chemie, Humboldt Universitaet zu
            Berlin, Invalidenstrasse 42, Berlin 10115, Germany"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     CDS             complement(140..157)
                     /label="6xHis"
                     /note="6xHis affinity tag"
     misc_feature    complement(158..203)
                     /label="multiple cloning site"
                     /note="multiple cloning site"
     CDS             complement(205..237)
                     /label="T7 tag (gene 10 leader)"
                     /note="leader peptide from bacteriophage T7 gene 10"
     CDS             complement(241..258)
                     /label="thrombin site"
                     /note="thrombin recognition and cleavage site"
     CDS             complement(268..285)
                     /label="6xHis"
                     /note="6xHis affinity tag"
     RBS             complement(304..326)
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      complement(339..387)
                     /label="lambda pR promoter"
                     /note="lambda pR promoter"
                     /regulatory_class="promoter"
     terminator      complement(402..429)
                     /label="T7Te terminator"
                     /note="phage T7 early transcription terminator"
     terminator      complement(445..516)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(564..596)
                     /label="ssrA tag (LVA)"
                     /note="C-terminal peptide that mediates degradation in
                     bacteria through the ClpXP and ClpAP proteases (McGinness
                     et al., 2006)"
     CDS             complement(597..1307)
                     /label="lambda repressor"
                     /note="phage lambda repressor"
     RBS             1314..1325
                     /note="strong bacterial ribosome binding site (Elowitz and
                     Leibler, 2000)"
     regulatory      complement(1332..1578)
                     /label="FixK2 promoter"
                     /note="FixK2 promoter"
                     /regulatory_class="promoter"
     promoter        1873..1950
                     /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             1951..3093
                     /codon_start=1
                     /product="YF1"
                     /label="YF1"
                     /protein_id="AFH34994.1"
                     /translation="MASFQSFGIPGQLEVIKKALDHVRVGVVITDPALEDNPIVYVNQG
                     FVQMTGYETEEILGKNCRFLQGKHTDPAEVDNIRTALQNKEPVTVQIQNYKKDGTMFWN
                     ELNIDPMEIEDKTYFVGIQNDITEHQQTQARLQELQSELVHVSRLSAMGEMASALAHEL
                     NQPLAAISNYMKGSRRLLAGSSDPNTPKVESALDRAAEQALRAGQIIRRLRDFVARGES
                     EKRVESLSKLIEEAGALGLAGAREQNVQLRFSLDPGADLVLADRVQIQQVLVNLFRNAL
                     EAMAQSQRRELVVTNTPAADDMIEVEVSDTGSGFQDDVIPNLFQTFFTTKDTGMGVGLS
                     ISRSIIEAHGGRMWAESNASGGATFRFTLPAADEMIGGLA"
     CDS             3090..3704
                     /gene="fixJ"
                     /label="Transcriptional regulatory protein FixJ"
                     /note="Transcriptional regulatory protein FixJ from
                     Bradyrhizobium diazoefficiens (strain JCM 10833 / BCRC
                     13528 / IAM 13628 / NBRC 14792 / USDA 110). Accession#:
                     P23221"
     CDS             4508..4696
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    4801..4943
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5129..5717)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             5839..6651
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(6747..7202)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
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 tttctagagc aaccattatc accgccagag
      361 gtaaaatagt caacacgcac ggtgttactc tagtatataa acgcagaaag gcccacccga
      421 aggtgagcca gtgtgactct agtagagagc gttcaccgac aaacaacaga taaaacgaaa
      481 ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggctctag tagcgatcta
      541 cactagcact atcagcgtta ttaagctact aaagcgtagt tttcgtcgtt tgcagcgcca
      601 aacgtctctt caggccactg actagcgata actttcccca caacggaaca actctcattg
      661 catgggatca ttgggtactg tgggtttagt ggttgtaaaa acacctgacc gctatccctg
      721 atcagtttct tgaaggtaaa ctcatcaccc ccaagtctgg ctatgcagaa atcacctggc
      781 tcaacagcct gctcagggtc aacgagaatt aacattccgt caggaaagct cggcttggag
      841 cctgttggtg cggtcatgga attaccttca acctcaagcc agaatgcaga atcactggct
      901 tttttggttg tgcttaccca tctctccgca tcacctttgg taaaggttct aagctcaggt
      961 gagaacatcc ctgcctgaac atgagaaaaa acagggtact catactcact tctaagtgac
     1021 ggctgcatac taaccgcttc atacatctcg tagatttctc tggcgattga agggctaaat
     1081 tcttcaacgc taactttgag aatttttgca agcaatgcgg cgttataagc atttaatgca
     1141 ttgatgccat taaataaagc accaacgcct gactgcccca tccccatctt gtctgcgaca
     1201 gattcctggg ataagccaag ttcatttttc tttttttcat aaattgcttt aaggcgacgt
     1261 gcgtcctcaa gctgctcttg tgttaatggt ttcttttttg tgctcatcta gtatttctcc
     1321 tcttttctag actccgttgt gatgacgcat tggtacgcgg tatcgggagg ttcgaaaatt
     1381 tcgagcgata tcttaagggg ggtgccttac gtagaacccc gtaggtcatg cccgaggccg
     1441 gtcctggatg gcgcggcgga tacgcttgag caggttttcg tcgagaagcg gcttcaaaac
     1501 cacgtctttt acgccggcct cggcggcccg ggtcgagatg ttttcgtccg gatagccggt
     1561 gatcaggatc acgggcgtag atctcgatcc tctacgccgg acgcatcgtg gccggcatca
     1621 ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc
     1681 gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg
     1741 tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc
     1801 tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc
     1861 gtcgagatcc cggacaccat cgaatggtgc aaaacctttc gcggtatggc atgatagcgc
     1921 ccggaagaga gtcaattgag ggtggtgaat gtggctagtt ttcaatcatt tgggatacca
     1981 ggacagctgg aagtcatcaa aaaagcactt gatcacgtgc gagtcggtgt ggtaattaca
     2041 gatcccgcac ttgaagataa tcctattgtc tacgtaaatc aaggctttgt tcaaatgacc
     2101 ggctacgaga ccgaggaaat tttaggaaag aactgtcgct tcttacaggg gaaacacaca
     2161 gatcctgcag aagtggacaa catcagaacc gctttacaaa ataaagaacc ggtcaccgtt
     2221 cagatccaaa actacaaaaa agacggaacg atgttctgga atgaattaaa tattgatcca
     2281 atggaaatag aggataaaac gtattttgtc ggtattcaga atgatatcac cgagcaccag
     2341 cagacccagg cgcgcctcca ggaactgcaa tccgagctcg tccacgtctc caggctgagc
     2401 gccatgggcg aaatggcgtc cgcgctcgcg cacgagctca accagccgct ggcggcgatc
     2461 agcaactaca tgaagggctc gcggcggctg cttgccggca gcagtgatcc gaacacaccg
     2521 aaggtcgaaa gcgccctgga ccgcgccgcc gagcaggcgc tgcgcgccgg ccagatcatc
     2581 cggcgcctgc gcgacttcgt tgcccgcggc gaatcggaga agcgggtcga gagtctctcc
     2641 aagctgatcg aggaggccgg cgcgctcggg cttgccggcg cgcgcgagca gaacgtgcag
     2701 ctccgcttca gtctcgatcc gggcgccgat ctcgttctcg ccgaccgggt gcagatccag
     2761 caggtcctgg tcaacctgtt ccgcaacgcg ctggaagcga tggctcagtc gcagcgacgc
     2821 gagctcgtcg tcaccaacac ccccgccgcc gacgacatga tcgaggtcga agtgtccgac
     2881 accggcagcg gtttccagga cgacgtcatt ccgaacctgt ttcagacttt cttcaccacc
     2941 aaggacaccg gcatgggcgt gggactgtcc atcagccgct cgatcatcga agctcacggc
     3001 gggcgcatgt gggccgagag caacgcatcg ggcggggcga ccttccgctt caccctcccg
     3061 gcagccgacg agatgatagg aggtctagca tgacgaccaa gggacatatc tacgtcatcg
     3121 acgacgacgc ggcgatgcgg gattcgctga atttcctgct ggattctgcc ggcttcggcg
     3181 tcacgctgtt tgacgacgcg caagcctttc tcgacgccct gccgggtctc tccttcggct
     3241 gcgtcgtctc cgacgtgcgc atgccgggcc ttgacggcat cgagctgttg aagcggatga
     3301 aggcgcagca aagccccttt ccgatcctca tcatgaccgg tcacggcgac gtgccgctcg
     3361 cggtcgaggc gatgaagtta ggggcggtgg actttctgga aaagcctttc gaggacgacc
     3421 gcctcaccgc catgatcgaa tcggcgatcc gccaggccga gccggccgcc aagagcgagg
     3481 ccgtcgcgca ggatatcgcc gcccgcgtcg cctcgttgag ccccagggag cgccaggtca
     3541 tggaagggct gatcgccggc ctttccaaca agctgatcgc ccgcgagtac gacatcagcc
     3601 cgcgcaccat cgaggtgtat cgggccaacg tcatgaccaa gatgcaggcc aacagccttt
     3661 cggagctggt tcgcctcgcg atgcgcgccg gcatgctcaa cgattgacaa ttgatgtaag
     3721 ttagctcact cattaggcac cgggatctcg accgatgccc ttgagagcct tcaacccagt
     3781 cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt
     3841 tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg
     3901 ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc
     3961 cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat
     4021 tatcgccggc atggcggccc cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg
     4081 gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac gcgagcgaac
     4141 gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa tggtcttcgg
     4201 tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg
     4261 gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg tattaacgaa
     4321 gcgctggcat tgaccctgag tgatttttct ctggtcccgc cgcatccata ccgccagttg
     4381 tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag
     4441 catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc cttacacgga
     4501 ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca
     4561 gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg cagacatctg
     4621 tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga
     4681 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     4741 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     4801 cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca
     4861 tcagagcaga ttgtactgag agtgcaccat atatgcggtg tgaaataccg cacagatgcg
     4921 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4981 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5041 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5101 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5161 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5221 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5281 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5341 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5401 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5461 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5521 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     5581 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5641 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5701 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5761 acgaaaactc acgttaaggg attttggtca tgaacaataa aactgtctgc ttacataaac
     5821 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc taggccgcga
     5881 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
     5941 caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg
     6001 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
     6061 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
     6121 tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga agaatatcct
     6181 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
     6241 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca
     6301 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
     6361 gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc
     6421 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     6481 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     6541 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     6601 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaagaatta
     6661 attcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6721 cacatttccc cgaaaagtgc cacctgaaat tgtaaacgtt aatattttgt taaaattcgc
     6781 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     6841 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     6901 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     6961 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     7021 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     7081 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     7141 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     7201 gtcccattcg cca
//
