LOCUS       40924_1754        6349 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector LIC-pDEST-LC2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6349)
  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 6349)
  AUTHORS   Dortay H, Mueller-Roeber B.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-FEB-2011) Molecular Biology, University of Potsdam, 
            Germany, Karl-Liebknecht-Str. 24-25, Haus 20, Potsdam, Brandenburg 
            14476, Germany
REFERENCE   3  (bases 1 to 6349)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6349)
  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 
            (12-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..6349
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    10..17
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     misc_feature    18..684
                     /label=LIC stuffer fragment
                     /note="LIC stuffer fragment"
     misc_feature    685..692
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     CDS             698..718
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     CDS             728..1711
                     /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             1718..1735
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     misc_feature    1736..1738
                     /label=stop codon (TAG)
                     /note="stop codon (TAG)"
     terminator      1803..1850
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(2216..2244)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     promoter        2357..2461
                     /label=AmpR promoter
     CDS             2462..3319
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3493..4081
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4267..4409)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4514..4702)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        6267..6285
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             6317..6339
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    6347..6349
                     /label=start codon
                     /note="start codon"
ORIGIN
        1 ggttcttctg tttaaacctc agtgtcatca ttcgaaagat tcgaaatatt tcccaaagaa
       61 agctcatggc ccaaccacaa cacaaacaaa ggagtaacgg cagcatgctc ccatgcgggg
      121 gaaagcagtt tttacagaaa tttgctatgg ctgacggaga aggagggctc atacccaaag
      181 ctgaaaaatt cttatgtgaa caagagaggg aaagaagtcc ttgtactgtg gggtattcat
      241 cacccgtcta acagtaagga gcaacagaat atctatcaga atgaaaatgc ttatgtctct
      301 gtagtgactt caaattataa caggagattt accccggaaa tagcagaaag acccaaagta
      361 agagatcaag ctgggaggat gaactattac tggaccttgc taaaacccgg agacacaata
      421 atatttgagg caaatggaaa tctaatagca ccaaggtatg ctttcgcact gagtagaggc
      481 tttgggtccg gcatcatcac ctcaaacgca tcaatgcatg agtgtaacac gaagtgtcaa
      541 acacccctgg gagctataaa cagcagtctc cctttccaga atatacaccc agtcacaata
      601 ggagagtgcc caaaatacgt caggagtgcc aaattgagga tggttacagg actaaggaac
      661 attccgtcca ttcaatccgc tagcgtttaa acagggcgag aacctgtatt ttcagggcgg
      721 cggcggcatg gctcgggacc ctctgccatt ctttccacct ctgtacctgg gcggccctga
      781 gattacaacc gagaactgcg agagagagcc tatccacatt cctgggtcca tccagccaca
      841 cggggctctg ctcacagctg acggccactc cggagaggtg ctccaagtgt ccctgaatgc
      901 cgctaccttc ctgggccagg agcctactgt gctgcggggg cagaccctgg ctgccctgct
      961 ccccgagcag tggccagccc tgcaggcagc cctgccccca ggatgtccag atgccctcca
     1021 atacagggcc accctcgact ggccagctgc tgggcacctc agcctgactg tgcatcgggt
     1081 ggctgaactc ctgatcctgg agttcgaacc taccgaggcc tgggacagca ttggccctca
     1141 cgccctgagg aacgccatgt ttgctctgga aagcgctcca aacctgcggg ctctggccga
     1201 agtcgcaaca caaactgtga gagaactgac tggcttcgat cgggtgatgc tgtacaaatt
     1261 tgcccctgac gccactggag agatgattgc tgaggccaga cgggagggca tgcaggcttt
     1321 tctgggccac aggtttcccg catcccacac ccctgcacaa gctagggccc tctacacaag
     1381 acacctgctc cggctgaccg cagacaccag ggctgcagca gtgcccctcg accccgtgct
     1441 gaatccccag acaaatgctc ctacacctct gggcggagct gtcctcagag ctacatcccc
     1501 aatgcacatg cagtacctga ggaatatggg agtgggctcc tccctgagcg tcagcgtcgt
     1561 ggtcggcggc cagctgtggg gactgattgt ctgccaccat cagacaccct acgtgctgcc
     1621 accagatctg cggaccaccc tggaggagct ggggagaaag ctgtccgggc aggtgcagag
     1681 gaaagaagcc ggaatggacg agctgtataa aggtacccac caccatcacc accactaggg
     1741 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     1801 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     1861 gaactatatc cggatatcca caggacgggt gtggtcgcca tgatcgcgta gtcgatagtg
     1921 gctccaagta gcgaagcgag caggactggg cggcggccaa agcggtcgga cagtgctccg
     1981 agaacgggtg cgcatagaaa ttgcatcaac gcatatagcg ctagcagcac gccatagtga
     2041 ctggcgatgc tgtcggaatg gacgatatcc cgcaagaggc ccggcagtac cggcataacc
     2101 aagcctatgc ctacagcatc cagggtgacg gtgccgagga tgacgatgag cgcattgtta
     2161 gatttcatac acggtgcctg actgcgttag caatttaact gtgataaact accgcattaa
     2221 agcttatcga tgataagctg tcaaacatga gaattcttga agacgaaagg gcctcgtgat
     2281 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     2341 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2401 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2461 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     2521 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     2581 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     2641 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     2701 ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     2761 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     2821 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     2881 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     2941 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3001 gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3061 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3121 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3181 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     3241 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3301 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     3361 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3421 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3481 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3541 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3601 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     3661 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3721 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3781 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3841 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3901 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3961 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4021 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4081 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4141 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     4201 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     4261 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatata
     4321 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
     4381 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     4441 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     4501 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     4561 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     4621 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     4681 cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg ttcatggggg
     4741 taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat gatgaacatg
     4801 cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg cgggaccaga
     4861 gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt gttccacagg
     4921 gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc gctgacttcc
     4981 gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt gctcaggtcg
     5041 cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct
     5101 aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc acgatcatgc
     5161 gcacccgtgg ccaggaccca acgctgcccg agatgcgccg cgtgcggctg ctggagatgg
     5221 cggacgcgat ggatatgttc tgccaagggt tggtttgcgc attcacagtt ctccgcaaga
     5281 attgattggc tccaattctt ggagtggtga atccgttagc gaggtgccgc cggcttccat
     5341 tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac gcggggaggc agacaaggta
     5401 tagggcggcg cctacaatcc atgccaaccc gttccatgtg ctcgccgagg cggcataaat
     5461 cgccgtgacg atcagcggtc cagtgatcga agttaggctg gtaagagccg cgagcgatcc
     5521 ttgaagctgt ccctgatggt cgtcatctac ctgcctggac agcatggcct gcaacgcggg
     5581 catcccgatg ccgccggaag cgagaagaat cataatgggg aaggccatcc agcctcgcgt
     5641 cgcgaacgcc agcaagacgt agcccagcgc gtcggccgcc atgccggcga taatggcctg
     5701 cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa
     5761 gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc
     5821 gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac
     5881 agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
     5941 gaaggctctc aagggcatcg gtcgatcgac gctctccctt atgcgactcc tgcattagga
     6001 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
     6061 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     6121 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
     6181 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
     6241 ggatcgagat ctcgatcccg cgaaattaat acgactcact atagggagac cacaacggtt
     6301 tccctctaga aataattttg tttaacttta agaaggagat atacatatg
//
