LOCUS       40924_1794        4938 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector LIC-pPICZ-LC2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4938)
  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 4938)
  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 4938)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4938)
  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..4938
                     /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"
     misc_feature    957..964
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     misc_feature    965..1631
                     /label=LIC stuffer fragment
                     /note="LIC stuffer fragment"
     misc_feature    1632..1639
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     CDS             1645..1665
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     CDS             1675..2658
                     /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             2665..2682
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     misc_feature    2683..2685
                     /label=stop codon (TGA)
                     /note="stop codon (TGA)"
     terminator      2762..3008
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        3023..3434
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3442..3489
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             3508..3879
                     /codon_start=1
                     /label=BleoR
                     /note="antibiotic-binding protein"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      3948..4195
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      complement(4270..4858)
                     /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 aacgatgggt tcttctgttt
      961 aaacctcagt gtcatcattc gaaagattcg aaatatttcc caaagaaagc tcatggccca
     1021 accacaacac aaacaaagga gtaacggcag catgctccca tgcgggggaa agcagttttt
     1081 acagaaattt gctatggctg acggagaagg agggctcata cccaaagctg aaaaattctt
     1141 atgtgaacaa gagagggaaa gaagtccttg tactgtgggg tattcatcac ccgtctaaca
     1201 gtaaggagca acagaatatc tatcagaatg aaaatgctta tgtctctgta gtgacttcaa
     1261 attataacag gagatttacc ccggaaatag cagaaagacc caaagtaaga gatcaagctg
     1321 ggaggatgaa ctattactgg accttgctaa aacccggaga cacaataata tttgaggcaa
     1381 atggaaatct aatagcacca aggtatgctt tcgcactgag tagaggcttt gggtccggca
     1441 tcatcacctc aaacgcatca atgcatgagt gtaacacgaa gtgtcaaaca cccctgggag
     1501 ctataaacag cagtctccct ttccagaata tacacccagt cacaatagga gagtgcccaa
     1561 aatacgtcag gagtgccaaa ttgaggatgg ttacaggact aaggaacatt ccgtccattc
     1621 aatccgctag cgtttaaaca gggcgagaac ctgtattttc agggcggcgg cggcatggct
     1681 cgggaccctc tgccattctt tccacctctg tacctgggcg gccctgagat tacaaccgag
     1741 aactgcgaga gagagcctat ccacattcct gggtccatcc agccacacgg ggctctgctc
     1801 acagctgacg gccactccgg agaggtgctc caagtgtccc tgaatgccgc taccttcctg
     1861 ggccaggagc ctactgtgct gcgggggcag accctggctg ccctgctccc cgagcagtgg
     1921 ccagccctgc aggcagccct gcccccagga tgtccagatg ccctccaata cagggccacc
     1981 ctcgactggc cagctgctgg gcacctcagc ctgactgtgc atcgggtggc tgaactcctg
     2041 atcctggagt tcgaacctac cgaggcctgg gacagcattg gccctcacgc cctgaggaac
     2101 gccatgtttg ctctggaaag cgctccaaac ctgcgggctc tggccgaagt cgcaacacaa
     2161 actgtgagag aactgactgg cttcgatcgg gtgatgctgt acaaatttgc ccctgacgcc
     2221 actggagaga tgattgctga ggccagacgg gagggcatgc aggcttttct gggccacagg
     2281 tttcccgcat cccacacccc tgcacaagct agggccctct acacaagaca cctgctccgg
     2341 ctgaccgcag acaccagggc tgcagcagtg cccctcgacc ccgtgctgaa tccccagaca
     2401 aatgctccta cacctctggg cggagctgtc ctcagagcta catccccaat gcacatgcag
     2461 tacctgagga atatgggagt gggctcctcc ctgagcgtca gcgtcgtggt cggcggccag
     2521 ctgtggggac tgattgtctg ccaccatcag acaccctacg tgctgccacc agatctgcgg
     2581 accaccctgg aggagctggg gagaaagctg tccgggcagg tgcagaggaa agaagccgga
     2641 atggacgagc tgtataaagt cgaccatcat catcatcatc attgagtttg tagccttaga
     2701 catgactgtt cctcagttca agttgggcac ttacgagaag accggtcttg ctagattcta
     2761 atcaagagga tgtcagaatg ccatttgcct gagagatgca ggcttcattt ttgatacttt
     2821 tttatttgta acctatatag tataggattt tttttgtcat tttgtttctt ctcgtacgag
     2881 cttgctcctg atcagcctat ctcgcagctg atgaatatct tgtggtaggg gtttgggaaa
     2941 atcattcgag tttgatgttt ttcttggtat ttcccactcc tcttcagagt acagaagatt
     3001 aagtgagacc ttcgtttgtg cggatccccc acacaccata gcttcaaaat gtttctactc
     3061 cttttttact cttccagatt ttctcggact ccgcgcatcg ccgtaccact tcaaaacacc
     3121 caagcacagc atactaaatt ttccctcttt cttcctctag ggtgtcgtta attacccgta
     3181 ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt cgaaaaaggc
     3241 aataaaaatt tttatcacgt ttctttttct tgaaattttt ttttttagtt tttttctctt
     3301 tcagtgacct ccattgatat ttaagttaat aaacggtctt caatttctca agtttcagtt
     3361 tcatttttct tgttctatta caactttttt tacttcttgt tcattagaaa gaaagcatag
     3421 caatctaatc taaggggcgg tgttgacaat taatcatcgg catagtatat cggcatagta
     3481 taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc
     3541 accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg
     3601 gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc
     3661 gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg
     3721 gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg
     3781 ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg
     3841 gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtccg acggcggccc
     3901 acgggtccca ggcctcggag atccgtcccc cttttccttt gtcgatatca tgtaattagt
     3961 tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa aggaaggagt
     4021 tagacaacct gaagtctagg tccctattta tttttttata gttatgttag tattaagaac
     4081 gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg catgtaacat
     4141 tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggctttaa tttgcaagct
     4201 ggagaccaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4261 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4321 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4381 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4441 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
     4501 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4561 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4621 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4681 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     4741 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4801 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4861 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4921 gattttggtc atgagatc
//
