LOCUS       40924_1779        9305 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector LIC-pLEXSY-LC1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9305)
  AUTHORS   Dortay H, Akula UM, Westphal C, Sittig M, Mueller-Roeber B.
  TITLE     High-Throughput Protein Expression Using a Combination of 
            Ligation-Independent Cloning (LIC) and Infrared Fluorescent Protein 
            (IFP) Detection
  JOURNAL   PLoS ONE 6 (4), E18900 (2011)
  PUBMED    21541323
REFERENCE   2  (bases 1 to 9305)
  AUTHORS   Dortay H, Mueller-Roeber B.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-FEB-2011) Molecular Biology, University of Potsdam, 
            Germany, Karl-Liebknecht-Str. 24-25, Haus 20, Potsdam, Brandenburg 
            14476, Germany
REFERENCE   3  (bases 1 to 9305)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9305)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2011"; volume: "6"; issue: "4"; pages: "E18900"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (14-FEB-2011) Molecular Biology, University of Potsdam, Germany, 
            Karl-Liebknecht-Str. 24-25, Haus 20, Potsdam, Brandenburg 14476, 
            Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rRNA            1..686
                     /product="18S ribosomal RNA"
                     /label=5' SSU 18S rRNA
                     /note="5' SSU 18S rRNA"
     misc_feature    1094..1096
                     /label=start codon
                     /note="start codon"
     CDS             1106..1123
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             1133..2116
                     /codon_start=1
                     /label=IFP1.4
                     /note="bacteriophytochrome-based monomeric infrared
                     fluorescent protein (Shu et al., 2009)"
                     /translation="MARDPLPFFPPLYLGGPEITTENCEREPIHIPGSIQPHGALLTAD
                     GHSGEVLQVSLNAATFLGQEPTVLRGQTLAALLPEQWPALQAALPPGCPDALQYRATLD
                     WPAAGHLSLTVHRVAELLILEFEPTEAWDSIGPHALRNAMFALESAPNLRALAEVATQT
                     VRELTGFDRVMLYKFAPDATGEMIAEARREGMQAFLGHRFPASHTPAQARALYTRHLLR
                     LTADTRAAAVPLDPVLNPQTNAPTPLGGAVLRATSPMHMQYLRNMGVGSSLSVSVVVGG
                     QLWGLIVCHHQTPYVLPPDLRTTLEELGRKLSGQVQRKEAGMDELYK"
     CDS             2117..2137
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     misc_feature    2156..2163
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     misc_feature    2164..2830
                     /label=LIC stuffer fragment
                     /note="LIC stuffer fragment"
     misc_feature    2831..2838
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     CDS             4209..4733
                     /codon_start=1
                     /product="nourseothricin resistance marker"
                     /label=nourseothricin resistance marker
                     /protein_id="AEE81079.1"
                     /translation="MKISVIPEQVAETLDAENHFIVREVFDVHLSDQGFELSTRSVSPY
                     RKDYISDDDSDEDSACYGAFIDQELVGKIELNSTWNDLASIEHIVVSHTHRGKGVAHSL
                     IEFAKKWALSRQLLGIRLETQTNNVPACNLYAKCGFTLGGIDLFTYKTRPQVSNETAMY
                     WYWFSGAQDDA"
     rRNA            5357..6441
                     /product="18S ribosomal RNA"
                     /label=3' SSU 18S rRNA
                     /note="3' SSU 18S rRNA"
     promoter        complement(6459..6477)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6498..6514)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6522..6538)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(6546..6576)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6591..6612)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6900..7488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7662..8519)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8520..8624)
                     /label=AmpR promoter
     rep_origin      8651..9107
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     9248..9264
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        9274..9292
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aaattggata acttggcgaa acgccaagct aatacatgaa ccaaccgggt gttctccact
       61 ccagacggtg ggcaaccatc gtcgtgagac gcccagcgaa tgaatgacag taaaaccaat
      121 gccttcactg gcagtaacac ccagcagtgt tgactcaatt cattccgtgc gaaagccggc
      181 ttgttccggc gtcttttgac gaacaactgc cctatcagct ggtgatggcc gtgtagtgga
      241 ctgccatgca tggcgttgac gggagcgggg gattagggtt cgattccgga gagggagcct
      301 gagaaatagc taccacttct acggagggca gcaggcgcgc aaattgccca atgtcaaaac
      361 aaaacgatga ggcagcgaaa agaaatagag ttgtcagtcc atttggattg tcatttcaat
      421 gggggatatt taaacccatc caatatcgag taacaattgg aggacaagtc tggtgccagc
      481 acccgcggta attccagctc caaaagcgta tattaatgct gttgctgtta aagggttcgt
      541 agttgaactg tgggctgtgc aggtttgttc ctggtcgtcc cgtccatgtc ggatttggtg
      601 acccaggccc ttgcagcccg tgaacattca aagaaacaag aaacacggga gtggttcctt
      661 tcctgattta cgcatgtcat gcatgcgtcg aggatccagt cgcagcctga ccgcatcaca
      721 catcaaggcg ttacagcctc tccgtctttg ctgcacccaa gtgtattcgt gtgaccgcgt
      781 tgagggatct aatggattcg gaactttggt cgtttggtct ctcatgctgc ccaatcccat
      841 gcgcttttct ccgtgcctct ctgtctcccc ttccgttcta tttggacaac gatgtgtact
      901 gtaatgcgtg cgccatctga caacgaatag atcagcagca ttcgcacact tgcacatacc
      961 cagtgaagct tttgtgtctg tcgtattgac aacaccgact gcaacaaggt gtagatagaa
     1021 gttggccttc tcgctcgctc gcacgctctt cacgctcctg ctttccttgc tgtgccttgc
     1081 caccagatct gccatgggct ccggccatca ccatcaccat cacctcgaat caatggctcg
     1141 ggaccctctg ccattctttc cacctctgta cctgggcggc cctgagatta caaccgagaa
     1201 ctgcgagaga gagcctatcc acattcctgg gtccatccag ccacacgggg ctctgctcac
     1261 agctgacggc cactccggag aggtgctcca agtgtccctg aatgccgcta ccttcctggg
     1321 ccaggagcct actgtgctgc gggggcagac cctggctgcc ctgctccccg agcagtggcc
     1381 agccctgcag gcagccctgc ccccaggatg tccagatgcc ctccaataca gggccaccct
     1441 cgactggcca gctgctgggc acctcagcct gactgtgcat cgggtggctg aactcctgat
     1501 cctggagttc gaacctaccg aggcctggga cagcattggc cctcacgccc tgaggaacgc
     1561 catgtttgct ctggaaagcg ctccaaacct gcgggctctg gccgaagtcg caacacaaac
     1621 tgtgagagaa ctgactggct tcgatcgggt gatgctgtac aaatttgccc ctgacgccac
     1681 tggagagatg attgctgagg ccagacggga gggcatgcag gcttttctgg gccacaggtt
     1741 tcccgcatcc cacacccctg cacaagctag ggccctctac acaagacacc tgctccggct
     1801 gaccgcagac accagggctg cagcagtgcc cctcgacccc gtgctgaatc cccagacaaa
     1861 tgctcctaca cctctgggcg gagctgtcct cagagctaca tccccaatgc acatgcagta
     1921 cctgaggaat atgggagtgg gctcctccct gagcgtcagc gtcgtggtcg gcggccagct
     1981 gtggggactg attgtctgcc accatcagac accctacgtg ctgccaccag atctgcggac
     2041 caccctggag gagctgggga gaaagctgtc cgggcaggtg cagaggaaag aagccggaat
     2101 ggacgagctg tataaagaga acctgtattt tcagggcggc ggcatgggtt cttctgttta
     2161 aacctcagtg tcatcattcg aaagattcga aatatttccc aaagaaagct catggcccaa
     2221 ccacaacaca aacaaaggag taacggcagc atgctcccat gcgggggaaa gcagttttta
     2281 cagaaatttg ctatggctga cggagaagga gggctcatac ccaaagctga aaaattctta
     2341 tgtgaacaag agagggaaag aagtccttgt actgtggggt attcatcacc cgtctaacag
     2401 taaggagcaa cagaatatct atcagaatga aaatgcttat gtctctgtag tgacttcaaa
     2461 ttataacagg agatttaccc cggaaatagc agaaagaccc aaagtaagag atcaagctgg
     2521 gaggatgaac tattactgga ccttgctaaa acccggagac acaataatat ttgaggcaaa
     2581 tggaaatcta atagcaccaa ggtatgcttt cgcactgagt agaggctttg ggtccggcat
     2641 catcacctca aacgcatcaa tgcatgagtg taacacgaag tgtcaaacac ccctgggagc
     2701 tataaacagc agtctccctt tccagaatat acacccagtc acaataggag agtgcccaaa
     2761 atacgtcagg agtgccaaat tgaggatggt tacaggacta aggaacattc cgtccattca
     2821 atccgctagc gtttaaacag ggcgagaacc tgcggccgcc ctcctcctcc tttcttgttc
     2881 ctttcacgtc gccttctcgg ttgtagctgg cagacgacga gtcttacttt tacgtgtact
     2941 tctctataga tgatgtatga tctctctgca tgcgtgttcg tgcatgtgtc cgtgtgttgt
     3001 gtacgcgtgc gtctcgcctc agctctccgc gtgaaagggt ttgactgccc atgatgcgtg
     3061 tgtatatcca cgcgcaggca cgcacacaca cacacacaca cacacacaca ggcacacaca
     3121 ggcacacaaa cgcatctcag gccgagccgc atacgtctct cgcacggtct cgtttatttg
     3181 atcatgtagt tgagtattaa attgggaaga caaaaacata atagcacgaa gagtcgggca
     3241 cgaaaagccc gatctctctc tctctctctc gtgcgcgcag gggcgtgtgg gtgcacgacg
     3301 acgaggacgg aggggggaag ggaggcataa acggatcgat cccccatcgc acatgcgggt
     3361 acgagcatta tctgctgtgt ctggtccttt atcatatcgc tacccgcccg ccccccgccc
     3421 ccctcccccc cccccccccg cggctcctgt cactgtcgtt cctgcgactc cccacacacc
     3481 cgcgcaacgc tcatgtcacg aagagaaaag tagaaggcgt ggcgatgcgt tgcgcggccc
     3541 cctgtccgct ggcatgcaga cgtcgagccg tggaacggat ggtagcgaga gacaacgcgg
     3601 cgatgcagga aaggagattt actgcaggac gtctacacac gcgcgcacac cactgcgaat
     3661 ggcaccgtgc tgaggaaaat tgggggggag ggcgcacggg ggcggggcag gaaacggttg
     3721 gatcagcagc actctcaact tgtgtccgta ttgcgaagga gggagaggag cctgcccgta
     3781 ttatgggcgc tgaaacgtcg gaaccaccta tgaatcccac ttctgcgtcg cggaggcgta
     3841 ctgcttccga ccacctccgc agctcacgcg cgtcgagctc cctccctttc cccttctttc
     3901 cgttgtgtgt gtgtgtgtgt gtgtgtcttg tcacgatgca tggccatctc tcctccaacc
     3961 ctccacgcct tcctctcccc cgcccatcaa acgcgctacg gccacaacca tttcttcaag
     4021 tatcacatcc accaccactc ttacccacct accctcttcg actctgacac agccactgac
     4081 gccctctccg cctctctcgc tcgctgcaca ttgactctcc acagcccccc ctccctcgca
     4141 cagccagccc cacccaccca cccacccacc caccgctcta cacacctcca cagcagccag
     4201 atcctagtat gaagatttcg gtgatccctg agcaggtggc ggaaacattg gatgctgaga
     4261 accatttcat tgttcgtgaa gtgttcgatg tgcacctatc cgaccaaggc tttgaactat
     4321 ctaccagaag tgtgagcccc taccggaagg attacatctc ggatgatgac tctgatgaag
     4381 actctgcttg ctatggcgca ttcatcgacc aagagcttgt cgggaagatt gaactcaact
     4441 caacatggaa cgatctagcc tctatcgaac acattgttgt gtcgcacacg caccgaggca
     4501 aaggagtcgc gcacagtctc atcgaatttg cgaaaaagtg ggcactaagc agacagctcc
     4561 ttggcatacg attagagaca caaacgaaca atgtacctgc ctgcaatttg tacgcaaaat
     4621 gtggctttac tctcggcggc attgacctgt tcacgtataa aactagacct caagtctcga
     4681 acgaaacagc gatgtactgg tactggttct cgggagcaca ggatgacgcc taactagcct
     4741 cggagatcca ctagttctag ttctaggggc cgaattcaga tcctcgtgtg agcgttcgcg
     4801 gaatcggtcg ctcgtgttta tgcccgtctt ggtgttgtgc tcgcaaggcg gtgcagcagg
     4861 ataccgtcgc cctcctctct ccttgcttct ctgttcttca attcgcgatc tcacagaggc
     4921 cggctgtgca cgcccttcct caccctcctt ttcccacctc tcggccaccg gtcggctccg
     4981 ttccgtctgc cgtcgagaag ggacgggcat gtgcagctcc tccctttctc tcgcgcgcgc
     5041 atcttctctt gcttgtggca ctcacgctca tgcgtcaagg cggccccacg ccgagcccct
     5101 gcgctccctt ccctcttgcg catccgtagc gcggatgccg tcgatgcgca aggccggcat
     5161 gaaggagcgc gtgccctcaa gagggcacac tatcatgccc tacgtgggcc acgcagcgat
     5221 gaggccggct tcggcggaga tgcgtcacgc acgtgccaga tgatgcgcta cgccttcctt
     5281 gacttgcgcc cccctctctt cctccgtctc tcactctctc tctctcacac acacacacac
     5341 acacacacac acacaaagct ccggttcgtc cggccgtaac gccttttcaa ctcacggcct
     5401 ctaggaatga aggagggtag ttcgggggag aacgtactgg ggcgtcagag gtgaaattct
     5461 tagaccgcac caagacgaac tacagcgaag gcattcttca aggatacctt cctcaatcaa
     5521 gaaccaaagt gtggagatcg aagatgatta gagaccattg tagtccacac tgcaaacgat
     5581 gacacccatg aattggggat cttatgggcc ggcctgcggc agggtttacc ctgtgtcagc
     5641 accgcgcccg cttttaccaa cttacgtatc ttttctattc ggcctttacc ggccacccac
     5701 gggaatatcc tcagcacgtt ttctgttttt tcacgcgaaa gctttgaggt tacagtctca
     5761 ggggggagta cgttcgcaag agtgaaactt aaagaaattg acggaatggc accacaagac
     5821 gtggagcgtg cggtttaatt tgactcaaca cggggaactt taccagatcc ggacaggatg
     5881 aggattgaca gattgagtgt tctttctcga ttccctgaat ggtggtgcat ggccgctttt
     5941 ggtcggtgga gtgatttgtt tggttgattc cgtcaacgga cgagatccaa gctgcccagt
     6001 agaattcaga attgcccata gaatagcaaa ctcatcggcg ggttttaccc aacggtgggc
     6061 cgcattcggt cgaattcttc tctgcgggat tcctttgtaa ttgcacaagg tgaaattttg
     6121 ggcaacagca ggtctgtgat gctcctcaat gttctgggcg acacgcgcac tacaatgtca
     6181 gtgagaacaa gaaaaacgac ttttgtcgaa cctacttgat caaaagagtg gggaaacccc
     6241 ggaatcacat agacccactt gggaccgagg attgcaatta ttggtcgcgc aacgaggaat
     6301 gtctcgtagg cgcagctcat caaactgtgc cgattacgtc cctgccattt gtacacaccg
     6361 cccgtcgttg tttccgatga tggtgcaata caggtgatcg gacaggcggt gttttatccg
     6421 cccgaaagtt caccgatatt taaatccagc ttttgttccc tttagtgagg gttaattgcg
     6481 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     6541 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6601 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6661 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6721 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6781 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6841 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6901 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6961 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     7021 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     7081 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     7141 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     7201 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7261 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7321 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7381 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7441 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7501 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7561 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7621 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7681 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7741 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     7801 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7861 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7921 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7981 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     8041 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     8101 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     8161 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     8221 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     8281 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8341 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     8401 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     8461 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8521 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8581 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8641 gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     8701 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     8761 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggcg ccctttaggg
     8821 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     8881 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     8941 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     9001 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaaatg agctgattta
     9061 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcca ttcgccattc
     9121 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     9181 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     9241 cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg
     9301 gattt
//
