LOCUS       40924_28007        6580 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo410, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6580)
  AUTHORS   Scheller L, Strittmatter T, Fuchs D, Bojar D, Fussenegger M.
  TITLE     Generalized extracellular molecule sensor platform for programming 
            cellular behavior
  JOURNAL   Nat. Chem. Biol. 14 (7), 723-729 (2018)
  PUBMED    29686358
REFERENCE   2  (bases 1 to 6580)
  AUTHORS   Scheller L, Strittmatter T.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-NOV-2017) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 
            4058, Switzerland
REFERENCE   3  (bases 1 to 6580)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6580)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem. 
            Biol."; date: "2018"; volume: "14"; issue: "7"; pages: "723-729"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-NOV-2017) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 4058, 
            Switzerland"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    40..64
                     /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)."
     RBS             79..101
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     misc_feature    84..100
                     /label=T7_transl_en_RBS
                     /note="T7_transl_en_RBS"
     CDS             108..815
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     misc_feature    846..902
                     /label=GCN4 tag
                     /note="GCN4 tag"
     CDS             846..902
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     misc_feature    918..974
                     /label=GCN4 tag
                     /note="GCN4 tag"
     CDS             918..974
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     misc_feature    963..965
                     /label=R
                     /note="R"
     misc_feature    990..1046
                     /label=GCN4 tag
                     /note="GCN4 tag"
     CDS             990..1046
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     misc_feature    1062..1118
                     /label=GCN4 tag
                     /note="GCN4 tag"
     CDS             1062..1118
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     misc_feature    1134..1190
                     /label=GCN4 tag
                     /note="GCN4 tag"
     CDS             1134..1190
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     misc_feature    1206..1262
                     /label=GCN4 tag
                     /note="GCN4 tag"
     CDS             1206..1262
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     misc_feature    1278..1334
                     /label=GCN4 tag
                     /note="GCN4 tag"
     CDS             1278..1334
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     misc_feature    1350..1406
                     /label=GCN4 tag
                     /note="GCN4 tag"
     CDS             1350..1406
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             1425..1445
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     CDS             1461..1478
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      1545..1592
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1629..2084
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(2180..2992)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      3114..3702
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(3888..4030)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4135..4323)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(5098..5119)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(5135..6214)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(6215..6292)
                     /label=lacI promoter
ORIGIN
        1 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
       61 ttcccctcta gaaataattt tgtttaactt taagaaggag atatagcatg gtgagcaagg
      121 gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
      181 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
      241 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
      301 tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
      361 ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
      421 aggacggcgg cgtggtgacc gtgacccagg actcctccct ccaggacggc gagttcatct
      481 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
      541 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg
      601 agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca
      661 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg
      721 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
      781 gccactccac cggcggcatg gacgagctgt acaagggatc caacggtccg actgacgccg
      841 cggaagaaga acttttgagc aagaattatc atcttgagaa cgaagtggct cgtcttaaga
      901 aaggttctgg cagtggagaa gaactgcttt caaagaatta ccacctggaa aatgaggtag
      961 ctagactgaa aaaggggagc ggaagtgggg aggagttgct gagcaaaaat tatcatttgg
     1021 agaacgaagt agcacgacta aagaaagggt ccggatcggg tgaggagtta ctctcgaaaa
     1081 attatcatct cgaaaacgaa gtggctcggc taaaaaaggg cagtggttct ggagaagagc
     1141 tattatctaa aaactaccac ctcgaaaatg aggtggcacg cttaaaaaag ggaagtggca
     1201 gtggtgaaga gctactatcc aagaattatc atcttgagaa cgaggtagcg cgtttgaaga
     1261 agggttccgg ctcaggagag gaactgctct cgaagaacta tcatcttgaa aatgaggtcg
     1321 ctcgattaaa aaagggatcg ggcagtggtg aggaactact ttcaaagaat taccacctcg
     1381 aaaacgaagt agctcgatta aagaaaggtt cagggtcggg tgaagaaaac ctgtacttcc
     1441 aatccgcggc cgcactcgag caccaccacc accaccactg agatccggct gctaacaaag
     1501 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     1561 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg
     1621 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     1681 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     1741 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     1801 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     1861 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     1921 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     1981 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     2041 aaaatttaac gcgaatttta acaaactagt aacgtttaca atttcaggtg gcacttttcg
     2101 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2161 gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     2221 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     2281 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     2341 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     2401 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
     2461 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     2521 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
     2581 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatgtttt
     2641 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg
     2701 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     2761 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     2821 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
     2881 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     2941 tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc ataacacccc
     3001 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgaccaaaat cccttaacgt
     3061 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3121 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3181 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3241 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     3301 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3361 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3421 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3481 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3541 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3601 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3661 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3721 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3781 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3841 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     3901 tttctcctta cgcatctgtg cggtatttca caccgcatat atggtgcact ctcagtacaa
     3961 tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt
     4021 catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     4081 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     4141 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     4201 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     4261 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     4321 cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg
     4381 agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg
     4441 tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca
     4501 atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc
     4561 gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga
     4621 aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca
     4681 gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg
     4741 ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat
     4801 gccggcgata atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc
     4861 ttgagcgagg gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct
     4921 ccagcgaaag cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag
     4981 ttgcatgata aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg
     5041 gaaggagctg actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg
     5101 agtgagctaa cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5161 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5221 gcgccagggt ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg
     5281 cctggccctg agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat
     5341 cctgtttgat ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc
     5401 ccactaccga gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc
     5461 ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca
     5521 tttgcatggt ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg
     5581 gctgaatttg attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga
     5641 cagaacttaa tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct
     5701 ccacgcccag tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt
     5761 cagagacatc aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat
     5821 cctggtcatc cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt
     5881 gcaccgccgc tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg
     5941 cacccagttg atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg
     6001 ccagactgga ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca
     6061 cgcggttggg aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg
     6121 cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat
     6181 actctgcgac atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt
     6241 ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct
     6301 cgacgctctc ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg
     6361 ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg
     6421 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
     6481 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
     6541 ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
//
