LOCUS       40924_1759        5250 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector LIC-pIVEX-LC1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5250)
  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 5250)
  AUTHORS   Dortay H, Mueller-Roeber B.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-FEB-2011) Molecular Biology, University of Potsdam, 
            Germany, Karl-Liebknecht-Str. 24-25, Haus 20, Potsdam, Brandenburg 
            14476, Germany
REFERENCE   3  (bases 1 to 5250)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5250)
  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 
            (11-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..5250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     381..397
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        620..638
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             670..692
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    699..701
                     /label=start codon
                     /note="start codon"
     CDS             711..728
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             738..749
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     CDS             756..1739
                     /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             1740..1760
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     misc_feature    1779..1786
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     misc_feature    1787..2453
                     /label=LIC stuffer fragment
                     /note="LIC stuffer fragment"
     misc_feature    2454..2461
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     terminator      2567..2614
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(2980..3008)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     primer_bind     complement(3029..3045)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3053..3069)
                     /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(3077..3107)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3122..3143)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3431..4019)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4193..5050)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5051..5155)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
      241 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
      301 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      361 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagcttgcat gcaaggagat
      421 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
      481 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
      541 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga
      601 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
      661 agaaataatt ttgtttaact ttaagaagga gatataccat gtctggttct catcatcatc
      721 atcatcatag cagcggcatc gaaggccgcg gccgcatggc tcgggaccct ctgccattct
      781 ttccacctct gtacctgggc ggccctgaga ttacaaccga gaactgcgag agagagccta
      841 tccacattcc tgggtccatc cagccacacg gggctctgct cacagctgac ggccactccg
      901 gagaggtgct ccaagtgtcc ctgaatgccg ctaccttcct gggccaggag cctactgtgc
      961 tgcgggggca gaccctggct gccctgctcc ccgagcagtg gccagccctg caggcagccc
     1021 tgcccccagg atgtccagat gccctccaat acagggccac cctcgactgg ccagctgctg
     1081 ggcacctcag cctgactgtg catcgggtgg ctgaactcct gatcctggag ttcgaaccta
     1141 ccgaggcctg ggacagcatt ggccctcacg ccctgaggaa cgccatgttt gctctggaaa
     1201 gcgctccaaa cctgcgggct ctggccgaag tcgcaacaca aactgtgaga gaactgactg
     1261 gcttcgatcg ggtgatgctg tacaaatttg cccctgacgc cactggagag atgattgctg
     1321 aggccagacg ggagggcatg caggcttttc tgggccacag gtttcccgca tcccacaccc
     1381 ctgcacaagc tagggccctc tacacaagac acctgctccg gctgaccgca gacaccaggg
     1441 ctgcagcagt gcccctcgac cccgtgctga atccccagac aaatgctcct acacctctgg
     1501 gcggagctgt cctcagagct acatccccaa tgcacatgca gtacctgagg aatatgggag
     1561 tgggctcctc cctgagcgtc agcgtcgtgg tcggcggcca gctgtgggga ctgattgtct
     1621 gccaccatca gacaccctac gtgctgccac cagatctgcg gaccaccctg gaggagctgg
     1681 ggagaaagct gtccgggcag gtgcagagga aagaagccgg aatggacgag ctgtataaag
     1741 agaacctgta ttttcagggc ggcggcatgg gttcttctgt ttaaacctca gtgtcatcat
     1801 tcgaaagatt cgaaatattt cccaaagaaa gctcatggcc caaccacaac acaaacaaag
     1861 gagtaacggc agcatgctcc catgcggggg aaagcagttt ttacagaaat ttgctatggc
     1921 tgacggagaa ggagggctca tacccaaagc tgaaaaattc ttatgtgaac aagagaggga
     1981 aagaagtcct tgtactgtgg ggtattcatc acccgtctaa cagtaaggag caacagaata
     2041 tctatcagaa tgaaaatgct tatgtctctg tagtgacttc aaattataac aggagattta
     2101 ccccggaaat agcagaaaga cccaaagtaa gagatcaagc tgggaggatg aactattact
     2161 ggaccttgct aaaacccgga gacacaataa tatttgaggc aaatggaaat ctaatagcac
     2221 caaggtatgc tttcgcactg agtagaggct ttgggtccgg catcatcacc tcaaacgcat
     2281 caatgcatga gtgtaacacg aagtgtcaaa cacccctggg agctataaac agcagtctcc
     2341 ctttccagaa tatacaccca gtcacaatag gagagtgccc aaaatacgtc aggagtgcca
     2401 aattgaggat ggttacagga ctaaggaaca ttccgtccat tcaatccgct agcgtttaaa
     2461 cagggcgaga acctggatcc ggtaactaac taagatccgg taagatccgg ctgctaacaa
     2521 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     2581 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggata
     2641 tccacaggac gggtgtggtc gccatgatcg cgtagtcgat agtggctcca agtagcgaag
     2701 cgagcaggac tgggcggcgg ccaaagcggt cggacagtgc tccgagaacg ggtgcgcata
     2761 gaaattgcat caacgcatat agcgctagca gcacgccata gtgactggcg atgctgtcgg
     2821 aatggacgat atcccgcaag aggcccggca gtaccggcat aaccaagcct atgcctacag
     2881 catccagggt gacggtgccg aggatgacga tgagcgcatt gttagatttc atacacggtg
     2941 cctgactgcg ttagcaattt aactgtgata aactaccgca ttaaagctta tcgatgataa
     3001 gctgtcaaac atgagaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc
     3061 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     3121 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3181 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3241 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3301 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3361 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3421 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3481 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3541 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3601 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3661 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3721 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3781 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3841 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     3901 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3961 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4021 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4081 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4141 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4201 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4261 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4321 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4381 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4441 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4501 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4561 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4621 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4681 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4741 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4801 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4861 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4921 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4981 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5041 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5101 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5161 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     5221 aaaataggcg tatcacgagg ccctttcgtc
//
