LOCUS       40924_1749        6366 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector LIC-pDEST-LC1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6366)
  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 6366)
  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 6366)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6366)
  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..6366
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             10..27
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             37..1020
                     /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             1021..1041
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     misc_feature    1060..1067
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     misc_feature    1068..1734
                     /label=LIC stuffer fragment
                     /note="LIC stuffer fragment"
     misc_feature    1735..1742
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     terminator      1820..1867
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(2233..2261)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     promoter        2374..2478
                     /label=AmpR promoter
     CDS             2479..3336
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3510..4098
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4284..4426)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4531..4719)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        6284..6302
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             6334..6356
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    6364..6366
                     /label=start codon
                     /note="start codon"
ORIGIN
        1 tcgtactacc atcaccatca ccatcacctc gaatcaatgg ctcgggaccc tctgccattc
       61 tttccacctc tgtacctggg cggccctgag attacaaccg agaactgcga gagagagcct
      121 atccacattc ctgggtccat ccagccacac ggggctctgc tcacagctga cggccactcc
      181 ggagaggtgc tccaagtgtc cctgaatgcc gctaccttcc tgggccagga gcctactgtg
      241 ctgcgggggc agaccctggc tgccctgctc cccgagcagt ggccagccct gcaggcagcc
      301 ctgcccccag gatgtccaga tgccctccaa tacagggcca ccctcgactg gccagctgct
      361 gggcacctca gcctgactgt gcatcgggtg gctgaactcc tgatcctgga gttcgaacct
      421 accgaggcct gggacagcat tggccctcac gccctgagga acgccatgtt tgctctggaa
      481 agcgctccaa acctgcgggc tctggccgaa gtcgcaacac aaactgtgag agaactgact
      541 ggcttcgatc gggtgatgct gtacaaattt gcccctgacg ccactggaga gatgattgct
      601 gaggccagac gggagggcat gcaggctttt ctgggccaca ggtttcccgc atcccacacc
      661 cctgcacaag ctagggccct ctacacaaga cacctgctcc ggctgaccgc agacaccagg
      721 gctgcagcag tgcccctcga ccccgtgctg aatccccaga caaatgctcc tacacctctg
      781 ggcggagctg tcctcagagc tacatcccca atgcacatgc agtacctgag gaatatggga
      841 gtgggctcct ccctgagcgt cagcgtcgtg gtcggcggcc agctgtgggg actgattgtc
      901 tgccaccatc agacacccta cgtgctgcca ccagatctgc ggaccaccct ggaggagctg
      961 gggagaaagc tgtccgggca ggtgcagagg aaagaagccg gaatggacga gctgtataaa
     1021 gagaacctgt attttcaggg cggcggcatg ggttcttctg tttaaacctc agtgtcatca
     1081 ttcgaaagat tcgaaatatt tcccaaagaa agctcatggc ccaaccacaa cacaaacaaa
     1141 ggagtaacgg cagcatgctc ccatgcgggg gaaagcagtt tttacagaaa tttgctatgg
     1201 ctgacggaga aggagggctc atacccaaag ctgaaaaatt cttatgtgaa caagagaggg
     1261 aaagaagtcc ttgtactgtg gggtattcat cacccgtcta acagtaagga gcaacagaat
     1321 atctatcaga atgaaaatgc ttatgtctct gtagtgactt caaattataa caggagattt
     1381 accccggaaa tagcagaaag acccaaagta agagatcaag ctgggaggat gaactattac
     1441 tggaccttgc taaaacccgg agacacaata atatttgagg caaatggaaa tctaatagca
     1501 ccaaggtatg ctttcgcact gagtagaggc tttgggtccg gcatcatcac ctcaaacgca
     1561 tcaatgcatg agtgtaacac gaagtgtcaa acacccctgg gagctataaa cagcagtctc
     1621 cctttccaga atatacaccc agtcacaata ggagagtgcc caaaatacgt caggagtgcc
     1681 aaattgagga tggttacagg actaaggaac attccgtcca ttcaatccgc tagcgtttaa
     1741 acagggcgag aacctggatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     1801 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1861 ttttttgctg aaaggaggaa ctatatccgg atatccacag gacgggtgtg gtcgccatga
     1921 tcgcgtagtc gatagtggct ccaagtagcg aagcgagcag gactgggcgg cggccaaagc
     1981 ggtcggacag tgctccgaga acgggtgcgc atagaaattg catcaacgca tatagcgcta
     2041 gcagcacgcc atagtgactg gcgatgctgt cggaatggac gatatcccgc aagaggcccg
     2101 gcagtaccgg cataaccaag cctatgccta cagcatccag ggtgacggtg ccgaggatga
     2161 cgatgagcgc attgttagat ttcatacacg gtgcctgact gcgttagcaa tttaactgtg
     2221 ataaactacc gcattaaagc ttatcgatga taagctgtca aacatgagaa ttcttgaaga
     2281 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     2341 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2401 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2461 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     2521 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     2581 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     2641 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     2701 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     2761 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     2821 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     2881 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     2941 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3001 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     3061 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3121 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3181 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3241 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     3301 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     3361 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     3421 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3481 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3541 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3601 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     3661 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3721 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3781 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3841 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3901 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3961 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4021 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4081 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4141 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4201 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     4261 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     4321 atttcacacc gcatatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4381 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     4441 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4501 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4561 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     4621 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     4681 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
     4741 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     4801 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     4861 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     4921 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     4981 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     5041 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     5101 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     5161 caggagcacg atcatgcgca cccgtggcca ggacccaacg ctgcccgaga tgcgccgcgt
     5221 gcggctgctg gagatggcgg acgcgatgga tatgttctgc caagggttgg tttgcgcatt
     5281 cacagttctc cgcaagaatt gattggctcc aattcttgga gtggtgaatc cgttagcgag
     5341 gtgccgccgg cttccattca ggtcgaggtg gcccggctcc atgcaccgcg acgcaacgcg
     5401 gggaggcaga caaggtatag ggcggcgcct acaatccatg ccaacccgtt ccatgtgctc
     5461 gccgaggcgg cataaatcgc cgtgacgatc agcggtccag tgatcgaagt taggctggta
     5521 agagccgcga gcgatccttg aagctgtccc tgatggtcgt catctacctg cctggacagc
     5581 atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat aatggggaag
     5641 gccatccagc ctcgcgtcgc gaacgccagc aagacgtagc ccagcgcgtc ggccgccatg
     5701 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     5761 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     5821 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     5881 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     5941 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gatcgacgct ctcccttatg
     6001 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
     6061 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
     6121 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
     6181 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
     6241 gatgcgtccg gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata
     6301 gggagaccac aacggtttcc ctctagaaat aattttgttt aactttaaga aggagatata
     6361 catatg
//
