LOCUS       40924_28107        7755 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo669, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7755)
  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 7755)
  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 7755)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7755)
  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..7755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    8..216
                     /label=Psv40
                     /note="Psv40"
     promoter        19..215
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      66..201
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      241..250
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    247..309
                     /label=igk secretion signal
                     /note="igk secretion signal"
     misc_feature    247..295
                     /label=secrete
                     /note="secrete"
     misc_feature    310..1044
                     /label=scfv
                     /note="scfv"
     misc_feature    1060..1803
                     /label=mouse EpoR-D1_D2_TM
                     /note="mouse EpoR-D1_D2_TM"
     misc_feature    1333..1335
                     /label=F93A
                     /note="F93A"
     misc_feature    1735..1803
                     /label=transmembrane
                     /note="transmembrane"
     misc_feature    1804..3075
                     /label=mFGFR 405-end
                     /note="mFGFR 405-end"
     polyA_signal    3129..3353
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3399..3827
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3841..4170
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4237..5028
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    5205..5338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5375..5391)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5399..5415)
                     /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(5423..5453)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5468..5489)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5777..6362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6536..7393)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7394..7498)
                     /label=AmpR promoter
ORIGIN
        1 tacgcgtgat ctgcgatctg catctcaatt agtcagcaac catagtcccg cccctaactc
       61 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat cgctgactaa
      121 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
      181 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tcgaatcgcg ctagcggccg
      241 gccaccatgg aaactgatac tttgctgctc tgggtcctgc tgctgtgggt ccctggatca
      301 acgggggacg gatccgacat cgtgatgacc cagagcccca gcagcctgag cgccagcgtg
      361 ggcgaccgcg tgaccatcac ctgccgcagc agcaccggcg ccgtgaccac cagcaactac
      421 gccagctggg tgcaggagaa gcccggcaag ctgttcaagg gcctgatcgg cggcaccaac
      481 aaccgcgccc ccggcgtgcc cagccgcttc agcggcagcc tgatcggcga caaggccacc
      541 ctgaccatca gcagcctgca gcccgaggac ttcgccacct acttctgcgc cctgtggtac
      601 agcaaccact gggtgttcgg ccagggcacc aaggtggagc tgaagcgcgg cggcggcggc
      661 agcggcggcg gcggcagcgg cggcggcggc agcagcggcg gcggcagcga ggtgaagctg
      721 ctggagagcg gcggcggcct ggtgcagccc ggcggcagcc tgaagctgag ctgcgccgtg
      781 agcggcttca gcctgaccga ctacggcgtg aactgggtgc gccaggcccc cggccgcggc
      841 ctggagtgga tcggcgtgat ctggggcgac ggcatcaccg actacaacag cgccctgaag
      901 gaccgcttca tcatcagcaa ggacaacggc aagaacaccg tgtacctgca gatgagcaag
      961 gtgcgcagcg acgacaccgc cctgtactac tgcgtgaccg gcctgttcga ctactggggc
     1021 cagggcaccc tggtgaccgt gagcagctcc ggagaattcg caccttcacc cagcctcccg
     1081 gaccccaagt ttgagagcaa agcggccctg ctggcatccc ggggctccga agaacttctg
     1141 tgcttcaccc aacgcttgga agacttggtg tgtttctggg aggaagcggc gagctccggg
     1201 atggacttca actacagctt ctcataccag ctcgagggtg agtcacgaaa gtcatgtagc
     1261 ctgcaccagg ctcccaccgt ccgcggctcc gtgcgtttct ggtgttcact gccaacagcg
     1321 gacacatcga gtgctgtgcc gctggagctg caggtgacgg aggcgtccgg ttctcctcgc
     1381 tatcaccgca tcatccatat caatgaagta gtgctcctgg acgcccccgc ggggctgctg
     1441 gcgcgccggg cagaagaggg cagccacgtg gtgctgcgct ggctgccacc tcctggagca
     1501 cctatgacca cccacatccg atatgaagtg gacgtgtcgg caggcaaccg ggcaggaggg
     1561 acacaaaggg tggaggtcct ggaaggccgc actgagtgtg ttctgagcaa cctgcggggc
     1621 gggacgcgct acaccttcgc tgttcgagcg cgcatggccg agccgagctt cagcggattc
     1681 tggagtgcct ggtctgagcc cgcgtcacta ctgaccgcta gcgacctgga ccctctcatc
     1741 ttgacgctgt ctctcattct ggtcctcatc tcgttgttgc tgacggttct ggccctgctg
     1801 tccatgaaga gcggcaccaa gaagagcgac ttccatagcc agatggctgt gcacaagctg
     1861 gccaagagca tccctctgcg cagacaggta acagtgtcag ctgactccag tgcatccatg
     1921 aactctgggg ttctcctggt tcggccctca cggctctcct ccagcgggac ccccatgctg
     1981 gctggagtct ccgaatatga gctccctgag gatccccgct gggagctgcc acgagacaga
     2041 ctggtcttag gcaaaccact tggcgagggc tgcttcgggc aggtggtgtt ggctgaggcc
     2101 atcgggctgg ataaggacaa acccaaccgt gtgaccaaag tggccgtgaa gatgttgaag
     2161 tccgacgcaa cggagaagga cctgtcggat ctgatctcgg agatggagat gatgaaaatg
     2221 attgggaagc acaagaatat catcaacctt ctgggagcgt gcacacagga tggtcctctt
     2281 tatgtcattg tggagtacgc ctccaaaggc aatctccggg agtatctaca ggcccggagg
     2341 cctcctgggc tggagtactg ctataacccc agccacaacc ccgaggaaca gctgtcttcc
     2401 aaagatctgg tatcctgtgc ctatcaggtg gctcggggca tggagtatct tgcctctaag
     2461 aagtgtatac accgagacct ggctgctagg aacgtcctgg tgaccgagga taacgtaatg
     2521 aagatcgcag actttggctt agctcgagac attcatcata tcgactacta caagaaaacc
     2581 accaacggcc ggctgcctgt gaagtggatg gcccctgagg cgttgtttga ccggatctac
     2641 acacaccaga gcgatgtgtg gtcttttgga gtgctcttgt gggagatctt cactctgggt
     2701 ggctccccat accccggtgt gcctgtggag gaacttttca agctgctgaa ggagggtcat
     2761 cgaatggaca agcccagtaa ctgtaccaat gagctgtaca tgatgatgcg ggactgctgg
     2821 catgcagtgc cctctcagag acctacgttc aagcagttgg tggaagacct ggaccgcatt
     2881 gtggccttga cctccaacca ggagtatctg gacctgtcca taccgctgga ccagtactca
     2941 cccagctttc ccgacacacg gagctccacc tgctcctcag gggaggactc tgtcttctct
     3001 catgagccgt tacctgagga gccctgtctg cctcgacacc ccacccagct tgccaacagt
     3061 ggactcaaac ggcgctaggc ggccgctcga gtctagaggg cccgtttaaa cccgctgatc
     3121 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3181 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3241 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3301 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     3361 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3421 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3481 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3541 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3601 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3661 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3721 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     3781 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     3841 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3901 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3961 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4021 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4081 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4141 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4201 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4261 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4321 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4381 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4441 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4501 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4561 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4621 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4681 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4741 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     4801 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4861 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4921 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4981 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5041 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5101 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5161 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     5221 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5281 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     5341 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5401 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5461 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5521 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5581 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5641 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5701 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5761 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5821 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5881 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5941 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6001 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6061 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6121 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6181 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6241 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6301 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6361 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6421 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6481 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6541 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6601 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6661 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6721 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6781 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6841 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6901 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6961 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7021 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7081 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7141 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7201 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7261 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7321 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7381 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7441 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7501 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     7561 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     7621 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     7681 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     7741 tgtacgggcc agata
//
