LOCUS       40924_28137        8969 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo694, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8969)
  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 8969)
  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 8969)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8969)
  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..8969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..1184
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     misc_feature    1200..1204
                     /label=SfiI
                     /note="SfiI"
     regulatory      1237..1246
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1243..1305
                     /label=igk secretion signal
                     /note="igk secretion signal"
     misc_feature    1306
                     /label=scfv
                     /note="scfv"
     misc_feature    1312..1689
                     /label=anti RR120
                     /note="anti RR120"
     misc_feature    1702..2445
                     /label=mouse EpoR-D1_D2_TM
                     /note="mouse EpoR-D1_D2_TM"
     misc_feature    1975..1977
                     /label=F93A
                     /note="F93A"
     misc_feature    2446..3717
                     /label=mFGFR 405-end
                     /note="mFGFR 405-end"
     polyA_signal    3812..4036
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    complement(4085..4104)
                     /label=itr
                     /note="itr"
     promoter        4468..4572
                     /label=AmpR promoter
     CDS             4573..5430
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5604..6192
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    6619..6638
                     /label=itr
                     /note="itr"
     polyA_signal    complement(6755..6810)
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     CDS             complement(6978..7574)
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    complement(7575..7634)
                     /label=P2A
                     /note="P2A"
     CDS             complement(7575..7631)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(7641..8339)
                     /label=TagBFP
                     /note="monomeric blue fluorescent protein"
     regulatory      complement(8353..8964)
                     /label=RPBSA
                     /note="RPBSA"
                     /regulatory_class="promoter"
     regulatory      8353..8964
                     /label=RPBSA
                     /note="RPBSA"
                     /regulatory_class="promoter"
ORIGIN
        1 aggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
       61 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      121 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      181 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      241 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      301 tacttccacg cccctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg
      361 ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg
      421 cttggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg
      481 ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt
      541 tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg
      601 gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc
      661 tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg
      721 tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg
      781 caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat
      841 ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct
      901 ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc
      961 tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg
     1021 cgatggagtt tccccatact gagtgggtgg agactgaagt taggccagct tggcacttga
     1081 tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc
     1141 agacagtggt tcaaagtttt tttcttccat ttaaggtgtc gtgaaaacta ccccaagctg
     1201 gcctctgagg ccagcttcga atcgcgctag cggccggcca ccatggaaac tgatactttg
     1261 ctgctctggg tcctgctgct gtgggtccct ggatcaacgg gggacggatc cgtgcagttg
     1321 caagaatcgg gtggtggact ggttcaagcc ggtgatagtt tgaaactgtc atgcgaagcc
     1381 agtggcgatt ccattggaac ttatgtcatt ggttggtttc gccaagcacc ggggaaagag
     1441 cgcatttacc ttgccactat tggtcgtaac ctggtcgggc cgtccgattt ttatacacgc
     1501 tacgctgact ctgtaaaggg tcgcttcgca gtgagtcgtg acaacgccaa gaacactgta
     1561 aacttacaga tgaactctct taaacctgaa gatactgctg tttactattg tgctgcgaaa
     1621 acgacaacgt gggggggaaa cgatcctaat aactggaatt attggggaca aggcactcag
     1681 gtgaccgtat ccggagaatt cgcaccttca cccagcctcc cggaccccaa gtttgagagc
     1741 aaagcggccc tgctggcatc ccggggctcc gaagaacttc tgtgcttcac ccaacgcttg
     1801 gaagacttgg tgtgtttctg ggaggaagcg gcgagctccg ggatggactt caactacagc
     1861 ttctcatacc agctcgaggg tgagtcacga aagtcatgta gcctgcacca ggctcccacc
     1921 gtccgcggct ccgtgcgttt ctggtgttca ctgccaacag cggacacatc gagtgctgtg
     1981 ccgctggagc tgcaggtgac ggaggcgtcc ggttctcctc gctatcaccg catcatccat
     2041 atcaatgaag tagtgctcct ggacgccccc gcggggctgc tggcgcgccg ggcagaagag
     2101 ggcagccacg tggtgctgcg ctggctgcca cctcctggag cacctatgac cacccacatc
     2161 cgatatgaag tggacgtgtc ggcaggcaac cgggcaggag ggacacaaag ggtggaggtc
     2221 ctggaaggcc gcactgagtg tgttctgagc aacctgcggg gcgggacgcg ctacaccttc
     2281 gctgttcgag cgcgcatggc cgagccgagc ttcagcggat tctggagtgc ctggtctgag
     2341 cccgcgtcac tactgaccgc tagcgacctg gaccctctca tcttgacgct gtctctcatt
     2401 ctggtcctca tctcgttgtt gctgacggtt ctggccctgc tgtccatgaa gagcggcacc
     2461 aagaagagcg acttccatag ccagatggct gtgcacaagc tggccaagag catccctctg
     2521 cgcagacagg taacagtgtc agctgactcc agtgcatcca tgaactctgg ggttctcctg
     2581 gttcggccct cacggctctc ctccagcggg acccccatgc tggctggagt ctccgaatat
     2641 gagctccctg aggatccccg ctgggagctg ccacgagaca gactggtctt aggcaaacca
     2701 cttggcgagg gctgcttcgg gcaggtggtg ttggctgagg ccatcgggct ggataaggac
     2761 aaacccaacc gtgtgaccaa agtggccgtg aagatgttga agtccgacgc aacggagaag
     2821 gacctgtcgg atctgatctc ggagatggag atgatgaaaa tgattgggaa gcacaagaat
     2881 atcatcaacc ttctgggagc gtgcacacag gatggtcctc tttatgtcat tgtggagtac
     2941 gcctccaaag gcaatctccg ggagtatcta caggcccgga ggcctcctgg gctggagtac
     3001 tgctataacc ccagccacaa ccccgaggaa cagctgtctt ccaaagatct ggtatcctgt
     3061 gcctatcagg tggctcgggg catggagtat cttgcctcta agaagtgtat acaccgagac
     3121 ctggctgcta ggaacgtcct ggtgaccgag gataacgtaa tgaagatcgc agactttggc
     3181 ttagctcgag acattcatca tatcgactac tacaagaaaa ccaccaacgg ccggctgcct
     3241 gtgaagtgga tggcccctga ggcgttgttt gaccggatct acacacacca gagcgatgtg
     3301 tggtcttttg gagtgctctt gtgggagatc ttcactctgg gtggctcccc ataccccggt
     3361 gtgcctgtgg aggaactttt caagctgctg aaggagggtc atcgaatgga caagcccagt
     3421 aactgtacca atgagctgta catgatgatg cgggactgct ggcatgcagt gccctctcag
     3481 agacctacgt tcaagcagtt ggtggaagac ctggaccgca ttgtggcctt gacctccaac
     3541 caggagtatc tggacctgtc cataccgctg gaccagtact cacccagctt tcccgacaca
     3601 cggagctcca cctgctcctc aggggaggac tctgtcttct ctcatgagcc gttacctgag
     3661 gagccctgtc tgcctcgaca ccccacccag cttgccaaca gtggactcaa acggcgctag
     3721 gcggccgctc gagtctagag ggcccgttta aacccggcct gtcaggccaa gcttggggat
     3781 caattctcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3841 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3901 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3961 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     4021 gcggtgggct ctatggcttc tgaggcggaa agaaccagct gggcctacta gatagcggac
     4081 cccttaccga aacatcgccg cattctgcag aggagtcgaa cttagtgtat gtaaacttct
     4141 gacccactgg aattgtgata cagtgaatta taagtgaaat aatctgtctg taaacaattg
     4201 ttggaaaaat gacttgtgtc atgcacaaag tagatgtcct aactgacttg ccaaaactat
     4261 tgtttgttaa caagaaattt gtggagtagt tgaaaaacga gttttaatga ctccaactta
     4321 agtgtatgta aacttccgac ttcaactgta tagggatcca attttaacaa aatattaacg
     4381 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4441 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4501 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4561 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4621 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4681 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4741 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4801 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4861 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4921 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4981 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5041 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5101 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5161 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5221 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5281 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5341 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5401 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5461 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5521 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5581 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5641 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5701 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5761 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5821 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5881 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5941 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6001 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6061 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6121 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6181 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6241 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6301 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6361 gaggaagcgg atccctatac agttgaagtc ggaagtttac atacacctta gccaaataca
     6421 tttaaactca ctttttcaca attcctgaca tttaatccta gtaaaaattc cctgtcttag
     6481 gtcagttagg atcaccactt tattttaaga atgtgaaata tcagaataat agtagagaga
     6541 atgattcatt tcagctttta tttctttcat cacattccca gtgggtcaga agtttacata
     6601 cactcgactc ctctgcagaa tgcggcgatg tttcggtaag gggtcccgag tagctagttc
     6661 atggcagcca gcatatggca tatgttgcca aactctaaac caaatactca ttctgatgtt
     6721 ttaaatgatt tgccctccca tatgtccttc cgagtgagag acacaaaaaa ttccaacaca
     6781 ctattgcaat gaaaataaat ttcctttatt agccagaagt cagatgctca aggggcttca
     6841 tgatgtcccc ataatttttg gcagagggaa aaagatctca gtggtatttg tgagccaggg
     6901 cattggccac accagccacc accttctgat aggcagcctg cacctgagga gtgaattcac
     6961 gacaggcctt cgaatcaggc accgggcttg cgggtcatgc accaggtgcg cggtccttcg
     7021 ggcacctcga cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg
     7081 ggcgcggagg tctccaggaa ggcgggcacc ccggcgcgct cggccgcctc cactccgggg
     7141 agcacgacgg cgctgcccag acccttgccc tggtggtcgg gcgagacgcc gacggtggcc
     7201 aggaaccacg cgggctcctt gggccggtgc ggcgccagga ggccttccat ctgttgctgc
     7261 gcggccagcc gggaaccgct caactcggcc atgcgcgggc cgatctcggc gaacaccgcc
     7321 cccgcttcga cgctctccgg cgtggtccag accgccaccg cggcgccgtc gtccgcgacc
     7381 cacaccttgc cgatgtcgag cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc
     7441 cgctcgatgt ggcggtccgg atcgacggtg tggcgcgtgg cggggtagtc ggcgaacgcg
     7501 gcggcgaggg tgcgtacggc cctggggacg tcgtcgcggg tggcgaggcg caccgtgggc
     7561 ttgtactcgg tcatggggcc aggattctcc tcgacgtcac cagcctgctt cagcaggctg
     7621 aagttagtag ctccactgcc gttcagcttg tgccccagtt tgctagggag gtcgcagtat
     7681 ctggccactg ccacctcgtg ctgctcgacg taggtctcgt tgttggcctc cttgattctt
     7741 tccagtctgt agtccacata gtagacgcca ggcatcttga ggttcttagc gggtttcttg
     7801 gatctatatg tggtcttgat gtttgcgatc agatggctcc cgcccacgag cttcagggcc
     7861 atgtcgtttc tgccttccag gccgccgtca gcggggtaca gcgtctcggt gaaggcctcc
     7921 cagccgagtg ttttcttctg catcacaggg ccgttggatg tgaagttcac ccctctgatc
     7981 ttgacgttgt agatgaggca gccgtcctgg aggctggtgt cctgggtagc ggtcagcacg
     8041 cccccgtctt cgtatgtggt gactctctcc catgtgaagc cctcagggaa ggactgcttg
     8101 aagaagtcgg ggatgccctg ggtgtggttg atgaaggtct tgctgccgta gaggaagcta
     8161 gtagccagga tgtcgaaggc gaaggggaga gggccgccct cgaccacctt gattctcatg
     8221 gtctgggtgc cctcgtaggg cttgccttcg ccctcggatg tgcacttgaa gtgatggttg
     8281 tccacggtgc cctccatgta cagcttcatg tgcatgttct ccttaatcag ctcgctcatg
     8341 gtggcctcag gtgcagaggt ttctacaggg aaagagagaa aggctacagt tagtttgctg
     8401 tgatgatgct aagtgccgag gtcagatgtc tatggattgt acacggaatc ccaacgcagg
     8461 aaccaaaaaa cactcaccca aaaaaatggc gctaaggccg agagaaaggc gaagttccgt
     8521 ctacggctat ataccttgaa acaggggcgg cagaagtgac gcaacgcggt tgcgcggacg
     8581 gcttcctgtg ttttcattgg tcgaagcgtt ctgcgcgtgc gcagagattc cccagggaag
     8641 gcgggccctc ttccgaatgt ccaatccgga cattcttccg gttggacctc cgcggtcagc
     8701 gctgtgcccg gcgagagatc acgtggggcg cggaggcggt gctgctgggg cacggccgtc
     8761 cagcctcggc ggccatattt ttgaggggct gttcatctcg ttcacacgct ctgtccgcca
     8821 tgtttgtgag tggaagcgcc attacccctt caagcgactg aaggctgcag ggcctctggt
     8881 ggcccgcatg gggagaccag acccgccagg cccgcctttc cgcactcagt ccgggcttac
     8941 tttattttgt gagacagggt ctcgcctag
//
