LOCUS       40924_1784        9288 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector LIC-pLEXSY-LC2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9288)
  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 9288)
  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 9288)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9288)
  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..9288
                     /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"
     misc_feature    1106..1113
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     misc_feature    1114..1780
                     /label=LIC stuffer fragment
                     /note="LIC stuffer fragment"
     misc_feature    1781..1788
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     CDS             1794..1814
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     CDS             1824..2807
                     /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             2814..2831
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     misc_feature    2832..2834
                     /label=stop codon (TAG)
                     /note="stop codon (TAG)"
     CDS             4192..4716
                     /codon_start=1
                     /product="nourseothricin resistance marker"
                     /label=nourseothricin resistance marker
                     /protein_id="AEE81081.1"
                     /translation="MKISVIPEQVAETLDAENHFIVREVFDVHLSDQGFELSTRSVSPY
                     RKDYISDDDSDEDSACYGAFIDQELVGKIELNSTWNDLASIEHIVVSHTHRGKGVAHSL
                     IEFAKKWALSRQLLGIRLETQTNNVPACNLYAKCGFTLGGIDLFTYKTRPQVSNETAMY
                     WYWFSGAQDDA"
     rRNA            5340..6424
                     /product="18S ribosomal RNA"
                     /label=3' SSU 18S rRNA
                     /note="3' SSU 18S rRNA"
     promoter        complement(6442..6460)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6481..6497)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6505..6521)
                     /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(6529..6559)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6574..6595)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6883..7471)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7645..8502)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8503..8607)
                     /label=AmpR promoter
     rep_origin      8634..9090
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     9231..9247
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        9257..9275
                     /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 gccatgggtt cttctgttta aacctcagtg tcatcattcg aaagattcga
     1141 aatatttccc aaagaaagct catggcccaa ccacaacaca aacaaaggag taacggcagc
     1201 atgctcccat gcgggggaaa gcagttttta cagaaatttg ctatggctga cggagaagga
     1261 gggctcatac ccaaagctga aaaattctta tgtgaacaag agagggaaag aagtccttgt
     1321 actgtggggt attcatcacc cgtctaacag taaggagcaa cagaatatct atcagaatga
     1381 aaatgcttat gtctctgtag tgacttcaaa ttataacagg agatttaccc cggaaatagc
     1441 agaaagaccc aaagtaagag atcaagctgg gaggatgaac tattactgga ccttgctaaa
     1501 acccggagac acaataatat ttgaggcaaa tggaaatcta atagcaccaa ggtatgcttt
     1561 cgcactgagt agaggctttg ggtccggcat catcacctca aacgcatcaa tgcatgagtg
     1621 taacacgaag tgtcaaacac ccctgggagc tataaacagc agtctccctt tccagaatat
     1681 acacccagtc acaataggag agtgcccaaa atacgtcagg agtgccaaat tgaggatggt
     1741 tacaggacta aggaacattc cgtccattca atccgctagc gtttaaacag ggcgagaacc
     1801 tgtattttca gggcggcggc ggcatggctc gggaccctct gccattcttt ccacctctgt
     1861 acctgggcgg ccctgagatt acaaccgaga actgcgagag agagcctatc cacattcctg
     1921 ggtccatcca gccacacggg gctctgctca cagctgacgg ccactccgga gaggtgctcc
     1981 aagtgtccct gaatgccgct accttcctgg gccaggagcc tactgtgctg cgggggcaga
     2041 ccctggctgc cctgctcccc gagcagtggc cagccctgca ggcagccctg cccccaggat
     2101 gtccagatgc cctccaatac agggccaccc tcgactggcc agctgctggg cacctcagcc
     2161 tgactgtgca tcgggtggct gaactcctga tcctggagtt cgaacctacc gaggcctggg
     2221 acagcattgg ccctcacgcc ctgaggaacg ccatgtttgc tctggaaagc gctccaaacc
     2281 tgcgggctct ggccgaagtc gcaacacaaa ctgtgagaga actgactggc ttcgatcggg
     2341 tgatgctgta caaatttgcc cctgacgcca ctggagagat gattgctgag gccagacggg
     2401 agggcatgca ggcttttctg ggccacaggt ttcccgcatc ccacacccct gcacaagcta
     2461 gggccctcta cacaagacac ctgctccggc tgaccgcaga caccagggct gcagcagtgc
     2521 ccctcgaccc cgtgctgaat ccccagacaa atgctcctac acctctgggc ggagctgtcc
     2581 tcagagctac atccccaatg cacatgcagt acctgaggaa tatgggagtg ggctcctccc
     2641 tgagcgtcag cgtcgtggtc ggcggccagc tgtggggact gattgtctgc caccatcaga
     2701 caccctacgt gctgccacca gatctgcgga ccaccctgga ggagctgggg agaaagctgt
     2761 ccgggcaggt gcagaggaaa gaagccggaa tggacgagct gtataaaggt acccaccacc
     2821 atcaccacca ctaggcggcc gccctcctcc tcctttcttg ttcctttcac gtcgccttct
     2881 cggttgtagc tggcagacga cgagtcttac ttttacgtgt acttctctat agatgatgta
     2941 tgatctctct gcatgcgtgt tcgtgcatgt gtccgtgtgt tgtgtacgcg tgcgtctcgc
     3001 ctcagctctc cgcgtgaaag ggtttgactg cccatgatgc gtgtgtatat ccacgcgcag
     3061 gcacgcacac acacacacac acacacacac acaggcacac acaggcacac aaacgcatct
     3121 caggccgagc cgcatacgtc tctcgcacgg tctcgtttat ttgatcatgt agttgagtat
     3181 taaattggga agacaaaaac ataatagcac gaagagtcgg gcacgaaaag cccgatctct
     3241 ctctctctct ctcgtgcgcg caggggcgtg tgggtgcacg acgacgagga cggagggggg
     3301 aagggaggca taaacggatc gatcccccat cgcacatgcg ggtacgagca ttatctgctg
     3361 tgtctggtcc tttatcatat cgctacccgc ccgccccccg cccccctccc cccccccccc
     3421 ccgcggctcc tgtcactgtc gttcctgcga ctccccacac acccgcgcaa cgctcatgtc
     3481 acgaagagaa aagtagaagg cgtggcgatg cgttgcgcgg ccccctgtcc gctggcatgc
     3541 agacgtcgag ccgtggaacg gatggtagcg agagacaacg cggcgatgca ggaaaggaga
     3601 tttactgcag gacgtctaca cacgcgcgca caccactgcg aatggcaccg tgctgaggaa
     3661 aattgggggg gagggcgcac gggggcgggg caggaaacgg ttggatcagc agcactctca
     3721 acttgtgtcc gtattgcgaa ggagggagag gagcctgccc gtattatggg cgctgaaacg
     3781 tcggaaccac ctatgaatcc cacttctgcg tcgcggaggc gtactgcttc cgaccacctc
     3841 cgcagctcac gcgcgtcgag ctccctccct ttccccttct ttccgttgtg tgtgtgtgtg
     3901 tgtgtgtgtc ttgtcacgat gcatggccat ctctcctcca accctccacg ccttcctctc
     3961 ccccgcccat caaacgcgct acggccacaa ccatttcttc aagtatcaca tccaccacca
     4021 ctcttaccca cctaccctct tcgactctga cacagccact gacgccctct ccgcctctct
     4081 cgctcgctgc acattgactc tccacagccc cccctccctc gcacagccag ccccacccac
     4141 ccacccaccc acccaccgct ctacacacct ccacagcagc cagatcctag tatgaagatt
     4201 tcggtgatcc ctgagcaggt ggcggaaaca ttggatgctg agaaccattt cattgttcgt
     4261 gaagtgttcg atgtgcacct atccgaccaa ggctttgaac tatctaccag aagtgtgagc
     4321 ccctaccgga aggattacat ctcggatgat gactctgatg aagactctgc ttgctatggc
     4381 gcattcatcg accaagagct tgtcgggaag attgaactca actcaacatg gaacgatcta
     4441 gcctctatcg aacacattgt tgtgtcgcac acgcaccgag gcaaaggagt cgcgcacagt
     4501 ctcatcgaat ttgcgaaaaa gtgggcacta agcagacagc tccttggcat acgattagag
     4561 acacaaacga acaatgtacc tgcctgcaat ttgtacgcaa aatgtggctt tactctcggc
     4621 ggcattgacc tgttcacgta taaaactaga cctcaagtct cgaacgaaac agcgatgtac
     4681 tggtactggt tctcgggagc acaggatgac gcctaactag cctcggagat ccactagttc
     4741 tagttctagg ggccgaattc agatcctcgt gtgagcgttc gcggaatcgg tcgctcgtgt
     4801 ttatgcccgt cttggtgttg tgctcgcaag gcggtgcagc aggataccgt cgccctcctc
     4861 tctccttgct tctctgttct tcaattcgcg atctcacaga ggccggctgt gcacgccctt
     4921 cctcaccctc cttttcccac ctctcggcca ccggtcggct ccgttccgtc tgccgtcgag
     4981 aagggacggg catgtgcagc tcctcccttt ctctcgcgcg cgcatcttct cttgcttgtg
     5041 gcactcacgc tcatgcgtca aggcggcccc acgccgagcc cctgcgctcc cttccctctt
     5101 gcgcatccgt agcgcggatg ccgtcgatgc gcaaggccgg catgaaggag cgcgtgccct
     5161 caagagggca cactatcatg ccctacgtgg gccacgcagc gatgaggccg gcttcggcgg
     5221 agatgcgtca cgcacgtgcc agatgatgcg ctacgccttc cttgacttgc gcccccctct
     5281 cttcctccgt ctctcactct ctctctctca cacacacaca cacacacaca cacacacaaa
     5341 gctccggttc gtccggccgt aacgcctttt caactcacgg cctctaggaa tgaaggaggg
     5401 tagttcgggg gagaacgtac tggggcgtca gaggtgaaat tcttagaccg caccaagacg
     5461 aactacagcg aaggcattct tcaaggatac cttcctcaat caagaaccaa agtgtggaga
     5521 tcgaagatga ttagagacca ttgtagtcca cactgcaaac gatgacaccc atgaattggg
     5581 gatcttatgg gccggcctgc ggcagggttt accctgtgtc agcaccgcgc ccgcttttac
     5641 caacttacgt atcttttcta ttcggccttt accggccacc cacgggaata tcctcagcac
     5701 gttttctgtt ttttcacgcg aaagctttga ggttacagtc tcagggggga gtacgttcgc
     5761 aagagtgaaa cttaaagaaa ttgacggaat ggcaccacaa gacgtggagc gtgcggttta
     5821 atttgactca acacggggaa ctttaccaga tccggacagg atgaggattg acagattgag
     5881 tgttctttct cgattccctg aatggtggtg catggccgct tttggtcggt ggagtgattt
     5941 gtttggttga ttccgtcaac ggacgagatc caagctgccc agtagaattc agaattgccc
     6001 atagaatagc aaactcatcg gcgggtttta cccaacggtg ggccgcattc ggtcgaattc
     6061 ttctctgcgg gattcctttg taattgcaca aggtgaaatt ttgggcaaca gcaggtctgt
     6121 gatgctcctc aatgttctgg gcgacacgcg cactacaatg tcagtgagaa caagaaaaac
     6181 gacttttgtc gaacctactt gatcaaaaga gtggggaaac cccggaatca catagaccca
     6241 cttgggaccg aggattgcaa ttattggtcg cgcaacgagg aatgtctcgt aggcgcagct
     6301 catcaaactg tgccgattac gtccctgcca tttgtacaca ccgcccgtcg ttgtttccga
     6361 tgatggtgca atacaggtga tcggacaggc ggtgttttat ccgcccgaaa gttcaccgat
     6421 atttaaatcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg
     6481 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6541 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6601 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6661 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6721 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6781 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6841 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6901 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6961 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7021 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7081 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7141 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7201 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7261 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7321 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7381 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7441 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7501 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7561 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     7621 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7681 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7741 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7801 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7861 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7921 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7981 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8041 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8101 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8161 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8221 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8281 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8341 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8401 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8461 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8521 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8581 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc
     8641 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     8701 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     8761 cggctttccc cgtcaagctc taaatcgggg gcgcccttta gggttccgat ttagtgcttt
     8821 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     8881 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     8941 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     9001 tttgccgatt tcggcctatt ggttaaaaaa atgagctgat ttaacaaaaa tttaacgcga
     9061 attttaacaa aatattaacg cttacaattt ccattcgcca ttcaggctgc gcaactgttg
     9121 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     9181 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     9241 ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttggattt
//
