LOCUS       40924_1764        5236 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector LIC-pIVEX-LC2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5236)
  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 5236)
  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 5236)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5236)
  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..5236
                     /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"
     misc_feature    711..718
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     misc_feature    719..1385
                     /label=LIC stuffer fragment
                     /note="LIC stuffer fragment"
     misc_feature    1386..1393
                     /label=PmeI recognition site
                     /note="PmeI recognition site"
     CDS             1399..1419
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     CDS             1429..2412
                     /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             2428..2445
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     misc_feature    2446..2451
                     /label=double stop codon (TAA TAA)
                     /note="double stop codon (TAA TAA)"
     terminator      2553..2600
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(2966..2994)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     primer_bind     complement(3015..3031)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3039..3055)
                     /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(3063..3093)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3108..3129)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3417..4005)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4179..5036)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5037..5141)
                     /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 gggttcttct gtttaaacct
      721 cagtgtcatc attcgaaaga ttcgaaatat ttcccaaaga aagctcatgg cccaaccaca
      781 acacaaacaa aggagtaacg gcagcatgct cccatgcggg ggaaagcagt ttttacagaa
      841 atttgctatg gctgacggag aaggagggct catacccaaa gctgaaaaat tcttatgtga
      901 acaagagagg gaaagaagtc cttgtactgt ggggtattca tcacccgtct aacagtaagg
      961 agcaacagaa tatctatcag aatgaaaatg cttatgtctc tgtagtgact tcaaattata
     1021 acaggagatt taccccggaa atagcagaaa gacccaaagt aagagatcaa gctgggagga
     1081 tgaactatta ctggaccttg ctaaaacccg gagacacaat aatatttgag gcaaatggaa
     1141 atctaatagc accaaggtat gctttcgcac tgagtagagg ctttgggtcc ggcatcatca
     1201 cctcaaacgc atcaatgcat gagtgtaaca cgaagtgtca aacacccctg ggagctataa
     1261 acagcagtct ccctttccag aatatacacc cagtcacaat aggagagtgc ccaaaatacg
     1321 tcaggagtgc caaattgagg atggttacag gactaaggaa cattccgtcc attcaatccg
     1381 ctagcgttta aacagggcga gaacctgtat tttcagggcg gcggcggcat ggctcgggac
     1441 cctctgccat tctttccacc tctgtacctg ggcggccctg agattacaac cgagaactgc
     1501 gagagagagc ctatccacat tcctgggtcc atccagccac acggggctct gctcacagct
     1561 gacggccact ccggagaggt gctccaagtg tccctgaatg ccgctacctt cctgggccag
     1621 gagcctactg tgctgcgggg gcagaccctg gctgccctgc tccccgagca gtggccagcc
     1681 ctgcaggcag ccctgccccc aggatgtcca gatgccctcc aatacagggc caccctcgac
     1741 tggccagctg ctgggcacct cagcctgact gtgcatcggg tggctgaact cctgatcctg
     1801 gagttcgaac ctaccgaggc ctgggacagc attggccctc acgccctgag gaacgccatg
     1861 tttgctctgg aaagcgctcc aaacctgcgg gctctggccg aagtcgcaac acaaactgtg
     1921 agagaactga ctggcttcga tcgggtgatg ctgtacaaat ttgcccctga cgccactgga
     1981 gagatgattg ctgaggccag acgggagggc atgcaggctt ttctgggcca caggtttccc
     2041 gcatcccaca cccctgcaca agctagggcc ctctacacaa gacacctgct ccggctgacc
     2101 gcagacacca gggctgcagc agtgcccctc gaccccgtgc tgaatcccca gacaaatgct
     2161 cctacacctc tgggcggagc tgtcctcaga gctacatccc caatgcacat gcagtacctg
     2221 aggaatatgg gagtgggctc ctccctgagc gtcagcgtcg tggtcggcgg ccagctgtgg
     2281 ggactgattg tctgccacca tcagacaccc tacgtgctgc caccagatct gcggaccacc
     2341 ctggaggagc tggggagaaa gctgtccggg caggtgcaga ggaaagaagc cggaatggac
     2401 gagctgtata aacccggggg gggttctcat catcatcatc atcattaata aaagggcgaa
     2461 ttccagcaca ctggcggccg ttactagtgg atccggctgc taacaaagcc cgaaaggaag
     2521 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     2581 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatatcca caggacgggt
     2641 gtggtcgcca tgatcgcgta gtcgatagtg gctccaagta gcgaagcgag caggactggg
     2701 cggcggccaa agcggtcgga cagtgctccg agaacgggtg cgcatagaaa ttgcatcaac
     2761 gcatatagcg ctagcagcac gccatagtga ctggcgatgc tgtcggaatg gacgatatcc
     2821 cgcaagaggc ccggcagtac cggcataacc aagcctatgc ctacagcatc cagggtgacg
     2881 gtgccgagga tgacgatgag cgcattgtta gatttcatac acggtgcctg actgcgttag
     2941 caatttaact gtgataaact accgcattaa agcttatcga tgataagctg tcaaacatga
     3001 gaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3061 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3121 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3181 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3241 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3301 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3361 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3421 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3481 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3541 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3601 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     3661 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     3721 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     3781 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     3841 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     3901 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     3961 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4021 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4081 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4141 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4201 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4261 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4321 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4381 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4441 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4501 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4561 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4621 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     4681 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     4741 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     4801 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     4861 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     4921 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     4981 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5041 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5101 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5161 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     5221 acgaggccct ttcgtc
//
