LOCUS       40924_1789        4982 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector LIC-pPICZ-LC1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4982)
  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 4982)
  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 4982)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4982)
  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..4982
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..935
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     misc_feature    945..947
                     /label=start codon
                     /note="start codon"
     CDS             957..974
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             984..1967
                     /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             1968..1988
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     misc_feature    2007..2014
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     misc_feature    2015..2681
                     /label=LIC stuffer fragment
                     /note="LIC stuffer fragment"
     misc_feature    2682..2689
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     CDS             2709..2726
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      2806..3052
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        3067..3478
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3486..3533
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             3552..3923
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      3992..4239
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(4314..4902)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag
       61 gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt
      121 tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc
      181 agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta
      241 acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta
      301 tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg
      361 agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaaggct
      421 gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg
      481 ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt
      541 cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct
      601 ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct
      661 ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact
      721 gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat
      781 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt
      841 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga
      901 caacttgaga agatcaaaaa acaactaatt attcgttcga aacgatgggc tccggccatc
      961 accatcacca tcacctcgaa tcaatggctc gggaccctct gccattcttt ccacctctgt
     1021 acctgggcgg ccctgagatt acaaccgaga actgcgagag agagcctatc cacattcctg
     1081 ggtccatcca gccacacggg gctctgctca cagctgacgg ccactccgga gaggtgctcc
     1141 aagtgtccct gaatgccgct accttcctgg gccaggagcc tactgtgctg cgggggcaga
     1201 ccctggctgc cctgctcccc gagcagtggc cagccctgca ggcagccctg cccccaggat
     1261 gtccagatgc cctccaatac agggccaccc tcgactggcc agctgctggg cacctcagcc
     1321 tgactgtgca tcgggtggct gaactcctga tcctggagtt cgaacctacc gaggcctggg
     1381 acagcattgg ccctcacgcc ctgaggaacg ccatgtttgc tctggaaagc gctccaaacc
     1441 tgcgggctct ggccgaagtc gcaacacaaa ctgtgagaga actgactggc ttcgatcggg
     1501 tgatgctgta caaatttgcc cctgacgcca ctggagagat gattgctgag gccagacggg
     1561 agggcatgca ggcttttctg ggccacaggt ttcccgcatc ccacacccct gcacaagcta
     1621 gggccctcta cacaagacac ctgctccggc tgaccgcaga caccagggct gcagcagtgc
     1681 ccctcgaccc cgtgctgaat ccccagacaa atgctcctac acctctgggc ggagctgtcc
     1741 tcagagctac atccccaatg cacatgcagt acctgaggaa tatgggagtg ggctcctccc
     1801 tgagcgtcag cgtcgtggtc ggcggccagc tgtggggact gattgtctgc caccatcaga
     1861 caccctacgt gctgccacca gatctgcgga ccaccctgga ggagctgggg agaaagctgt
     1921 ccgggcaggt gcagaggaaa gaagccggaa tggacgagct gtataaagag aacctgtatt
     1981 ttcagggcgg cggcatgggt tcttctgttt aaacctcagt gtcatcattc gaaagattcg
     2041 aaatatttcc caaagaaagc tcatggccca accacaacac aaacaaagga gtaacggcag
     2101 catgctccca tgcgggggaa agcagttttt acagaaattt gctatggctg acggagaagg
     2161 agggctcata cccaaagctg aaaaattctt atgtgaacaa gagagggaaa gaagtccttg
     2221 tactgtgggg tattcatcac ccgtctaaca gtaaggagca acagaatatc tatcagaatg
     2281 aaaatgctta tgtctctgta gtgacttcaa attataacag gagatttacc ccggaaatag
     2341 cagaaagacc caaagtaaga gatcaagctg ggaggatgaa ctattactgg accttgctaa
     2401 aacccggaga cacaataata tttgaggcaa atggaaatct aatagcacca aggtatgctt
     2461 tcgcactgag tagaggcttt gggtccggca tcatcacctc aaacgcatca atgcatgagt
     2521 gtaacacgaa gtgtcaaaca cccctgggag ctataaacag cagtctccct ttccagaata
     2581 tacacccagt cacaatagga gagtgcccaa aatacgtcag gagtgccaaa ttgaggatgg
     2641 ttacaggact aaggaacatt ccgtccattc aatccgctag cgtttaaaca gggcgagaac
     2701 ctgtcgacca tcatcatcat catcattgag tttgtagcct tagacatgac tgttcctcag
     2761 ttcaagttgg gcacttacga gaagaccggt cttgctagat tctaatcaag aggatgtcag
     2821 aatgccattt gcctgagaga tgcaggcttc atttttgata cttttttatt tgtaacctat
     2881 atagtatagg attttttttg tcattttgtt tcttctcgta cgagcttgct cctgatcagc
     2941 ctatctcgca gctgatgaat atcttgtggt aggggtttgg gaaaatcatt cgagtttgat
     3001 gtttttcttg gtatttccca ctcctcttca gagtacagaa gattaagtga gaccttcgtt
     3061 tgtgcggatc ccccacacac catagcttca aaatgtttct actccttttt tactcttcca
     3121 gattttctcg gactccgcgc atcgccgtac cacttcaaaa cacccaagca cagcatacta
     3181 aattttccct ctttcttcct ctagggtgtc gttaattacc cgtactaaag gtttggaaaa
     3241 gaaaaaagag accgcctcgt ttctttttct tcgtcgaaaa aggcaataaa aatttttatc
     3301 acgtttcttt ttcttgaaat tttttttttt agtttttttc tctttcagtg acctccattg
     3361 atatttaagt taataaacgg tcttcaattt ctcaagtttc agtttcattt ttcttgttct
     3421 attacaactt tttttacttc ttgttcatta gaaagaaagc atagcaatct aatctaaggg
     3481 gcggtgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
     3541 ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg
     3601 ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg
     3661 acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg
     3721 tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg
     3781 agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga
     3841 tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc
     3901 acttcgtggc cgaggagcag gactgacacg tccgacggcg gcccacgggt cccaggcctc
     3961 ggagatccgt cccccttttc ctttgtcgat atcatgtaat tagttatgtc acgcttacat
     4021 tcacgccctc cccccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc
     4081 taggtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt tatatttcaa
     4141 atttttcttt tttttctgta cagacgcgtg tacgcatgta acattatact gaaaaccttg
     4201 cttgagaagg ttttgggacg ctcgaaggct ttaatttgca agctggagac caacatgtga
     4261 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4321 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4381 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4441 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4501 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4561 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4621 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4681 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4741 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4801 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4861 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4921 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4981 tc
//
