LOCUS       40924_28142        8537 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo695, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8537)
  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 8537)
  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 8537)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8537)
  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..8537
                     /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"
     sig_peptide     1243..1305
                     /label=Ig-kappa leader
                     /note="leader sequence from mouse immunoglobulin kappa 
                     light
                     chain"
     misc_feature    1312..1689
                     /label=anti RR120
                     /note="anti RR120"
     misc_feature    1697..2445
                     /label=EpoR-D1_D2_TM
                     /note="EpoR-D1_D2_TM"
     misc_feature    1975..1977
                     /label=F93A
                     /note="F93A"
     misc_feature    2455..3283
                     /label=IL6ST
                     /note="IL6ST"
     misc_feature    2806..2808
                     /label=Y759A
                     /note="Y759A"
     polyA_signal    3380..3604
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    complement(3653..3672)
                     /label=itr
                     /note="itr"
     promoter        4036..4140
                     /label=AmpR promoter
     CDS             4141..4998
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5172..5760
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    6187..6206
                     /label=itr
                     /note="itr"
     polyA_signal    complement(6323..6378)
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     CDS             complement(6546..7142)
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    complement(7143..7202)
                     /label=P2A
                     /note="P2A"
     CDS             complement(7143..7199)
                     /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(7209..7907)
                     /label=TagBFP
                     /note="monomeric blue fluorescent protein"
     regulatory      complement(7921..8532)
                     /label=RPBSA
                     /note="RPBSA"
                     /regulatory_class="promoter"
     regulatory      7921..8532
                     /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 tgtccgcagc agcaaataag
     2461 cgagacctaa ttaaaaaaca catctggcct aatgttccag atccttcaaa gagtcatatt
     2521 gcccagtggt cacctcacac tcctccaagg cacaatttta attcaaaaga tcaaatgtat
     2581 tcagatggca atttcactga tgtaagtgtt gtggaaatag aagcaaatga caaaaagcct
     2641 tttccagaag atctgaaatc attggacctg ttcaaaaagg aaaaaattaa tactgaagga
     2701 cacagcagtg gtattggggg gtcttcatgc atgtcatctt ctaggccaag catttctagc
     2761 agtgatgaaa atgaatcttc acaaaacact tcgagcactg tccaggcttc taccgtggta
     2821 cacagtggct acagacacca agttccgtca gtccaagtct tctcaagatc cgagtctacc
     2881 cagcccttgt tagattcaga ggagcggcca gaagatctac aattagtaga tcatgtagat
     2941 ggcggtgatg gtattttgcc caggcaacag tacttcaaac agaactgcag tcagcatgaa
     3001 tccagtccag atatttcaca ttttgaaagg tcaaagcaag tttcatcagt caatgaggaa
     3061 gattttgtta gacttaaaca gcagatttca gatcatattt cacaatcctg tggatctggg
     3121 caaatgaaaa tgtttcagga agtttctgca gcagatgctt ttggtccagg tactgaggga
     3181 caagtagaaa gatttgaaac agttggcatg gaggctgcga ctgatgaagg catgcctaaa
     3241 agttacttac cacagactgt acggcaaggc ggctacatgc ctcagtaggc ggccgctcga
     3301 gtctagaggg cccgtttaaa cccggcctgt caggccaagc ttggggatca attctctaga
     3361 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     3421 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     3481 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     3541 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     3601 atggcttctg aggcggaaag aaccagctgg gcctactaga tagcggaccc cttaccgaaa
     3661 catcgccgca ttctgcagag gagtcgaact tagtgtatgt aaacttctga cccactggaa
     3721 ttgtgataca gtgaattata agtgaaataa tctgtctgta aacaattgtt ggaaaaatga
     3781 cttgtgtcat gcacaaagta gatgtcctaa ctgacttgcc aaaactattg tttgttaaca
     3841 agaaatttgt ggagtagttg aaaaacgagt tttaatgact ccaacttaag tgtatgtaaa
     3901 cttccgactt caactgtata gggatccaat tttaacaaaa tattaacgct tacaatttcc
     3961 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcatc aggtggcact
     4021 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4081 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4141 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4201 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4261 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4321 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4381 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4441 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     4501 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     4561 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     4621 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     4681 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     4741 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     4801 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     4861 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4921 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4981 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     5041 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5101 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5161 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5221 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5281 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5341 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5401 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5461 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5521 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5581 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5641 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5701 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5761 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5821 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     5881 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggat
     5941 ccctatacag ttgaagtcgg aagtttacat acaccttagc caaatacatt taaactcact
     6001 ttttcacaat tcctgacatt taatcctagt aaaaattccc tgtcttaggt cagttaggat
     6061 caccacttta ttttaagaat gtgaaatatc agaataatag tagagagaat gattcatttc
     6121 agcttttatt tctttcatca cattcccagt gggtcagaag tttacataca ctcgactcct
     6181 ctgcagaatg cggcgatgtt tcggtaaggg gtcccgagta gctagttcat ggcagccagc
     6241 atatggcata tgttgccaaa ctctaaacca aatactcatt ctgatgtttt aaatgatttg
     6301 ccctcccata tgtccttccg agtgagagac acaaaaaatt ccaacacact attgcaatga
     6361 aaataaattt cctttattag ccagaagtca gatgctcaag gggcttcatg atgtccccat
     6421 aatttttggc agagggaaaa agatctcagt ggtatttgtg agccagggca ttggccacac
     6481 cagccaccac cttctgatag gcagcctgca cctgaggagt gaattcacga caggccttcg
     6541 aatcaggcac cgggcttgcg ggtcatgcac caggtgcgcg gtccttcggg cacctcgacg
     6601 tcggcggtga cggtgaagcc gagccgctcg tagaagggga ggttgcgggg cgcggaggtc
     6661 tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca ctccggggag cacgacggcg
     6721 ctgcccagac ccttgccctg gtggtcgggc gagacgccga cggtggccag gaaccacgcg
     6781 ggctccttgg gccggtgcgg cgccaggagg ccttccatct gttgctgcgc ggccagccgg
     6841 gaaccgctca actcggccat gcgcgggccg atctcggcga acaccgcccc cgcttcgacg
     6901 ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt ccgcgaccca caccttgccg
     6961 atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct cggtgacccg ctcgatgtgg
     7021 cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg cgaacgcggc ggcgagggtg
     7081 cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca ccgtgggctt gtactcggtc
     7141 atggggccag gattctcctc gacgtcacca gcctgcttca gcaggctgaa gttagtagct
     7201 ccactgccgt tcagcttgtg ccccagtttg ctagggaggt cgcagtatct ggccactgcc
     7261 acctcgtgct gctcgacgta ggtctcgttg ttggcctcct tgattctttc cagtctgtag
     7321 tccacatagt agacgccagg catcttgagg ttcttagcgg gtttcttgga tctatatgtg
     7381 gtcttgatgt ttgcgatcag atggctcccg cccacgagct tcagggccat gtcgtttctg
     7441 ccttccaggc cgccgtcagc ggggtacagc gtctcggtga aggcctccca gccgagtgtt
     7501 ttcttctgca tcacagggcc gttggatgtg aagttcaccc ctctgatctt gacgttgtag
     7561 atgaggcagc cgtcctggag gctggtgtcc tgggtagcgg tcagcacgcc cccgtcttcg
     7621 tatgtggtga ctctctccca tgtgaagccc tcagggaagg actgcttgaa gaagtcgggg
     7681 atgccctggg tgtggttgat gaaggtcttg ctgccgtaga ggaagctagt agccaggatg
     7741 tcgaaggcga aggggagagg gccgccctcg accaccttga ttctcatggt ctgggtgccc
     7801 tcgtagggct tgccttcgcc ctcggatgtg cacttgaagt gatggttgtc cacggtgccc
     7861 tccatgtaca gcttcatgtg catgttctcc ttaatcagct cgctcatggt ggcctcaggt
     7921 gcagaggttt ctacagggaa agagagaaag gctacagtta gtttgctgtg atgatgctaa
     7981 gtgccgaggt cagatgtcta tggattgtac acggaatccc aacgcaggaa ccaaaaaaca
     8041 ctcacccaaa aaaatggcgc taaggccgag agaaaggcga agttccgtct acggctatat
     8101 accttgaaac aggggcggca gaagtgacgc aacgcggttg cgcggacggc ttcctgtgtt
     8161 ttcattggtc gaagcgttct gcgcgtgcgc agagattccc cagggaaggc gggccctctt
     8221 ccgaatgtcc aatccggaca ttcttccggt tggacctccg cggtcagcgc tgtgcccggc
     8281 gagagatcac gtggggcgcg gaggcggtgc tgctggggca cggccgtcca gcctcggcgg
     8341 ccatattttt gaggggctgt tcatctcgtt cacacgctct gtccgccatg tttgtgagtg
     8401 gaagcgccat taccccttca agcgactgaa ggctgcaggg cctctggtgg cccgcatggg
     8461 gagaccagac ccgccaggcc cgcctttccg cactcagtcc gggcttactt tattttgtga
     8521 gacagggtct cgcctag
//
