LOCUS       40924_28097        7371 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo667, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7371)
  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 7371)
  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 7371)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7371)
  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..7371
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    7..215
                     /label=Psv40
                     /note="Psv40"
     promoter        18..214
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      65..200
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      240..249
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    246..308
                     /label=igk secretion signal
                     /note="igk secretion signal"
     misc_feature    315..662
                     /label=Nic_VH
                     /note="Nic_VH"
     misc_feature    675..1418
                     /label=mouse EpoR-D1_D2_TM
                     /note="mouse EpoR-D1_D2_TM"
     misc_feature    948..950
                     /label=F93A
                     /note="F93A"
     misc_feature    1350..1418
                     /label=transmembrane
                     /note="transmembrane"
     misc_feature    1419..2690
                     /label=mFGFR 405-end
                     /note="mFGFR 405-end"
     polyA_signal    2744..2968
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3014..3442
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3456..3785
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3852..4643
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    4820..4953
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4990..5006)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5014..5030)
                     /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(5038..5068)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5083..5104)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5392..5977)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6151..7008)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7009..7113)
                     /label=AmpR promoter
ORIGIN
        1 acgcgtgatc tgcgatctgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
       61 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatc gctgactaat
      121 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
      181 aggaggcttt tttggaggcc taggcttttg caaaaagctt cgaatcgcgc tagcggccgg
      241 ccaccatgga aactgatact ttgctgctct gggtcctgct gctgtgggtc cctggatcaa
      301 cgggggacgg atcccagatg caactgttgg aatcgggacc gggtttggtc aagccgagcg
      361 agacattgag tttaacgtgc acggtttcag gcggatcaat ttggggttgg attcgccagc
      421 cgccaggaaa gggactggag tggattggat ctatttactc ctccggttcc acctactata
      481 atccgagctt gaagtcccgt gtgacaacaa gcgtggatac atccaaaaac cagttttcat
      541 tgcgcctgtc ctcagtaacc gcagccgaca cagccgtata ttattgcgtt gcttggtttg
      601 gcgacttatt aagtcttaaa ggggtagagc tttggggcca gggaactctt gtgacggtat
      661 cgtccggaga attcgcacct tcacccagcc tcccggaccc caagtttgag agcaaagcgg
      721 ccctgctggc atcccggggc tccgaagaac ttctgtgctt cacccaacgc ttggaagact
      781 tggtgtgttt ctgggaggaa gcggcgagct ccgggatgga cttcaactac agcttctcat
      841 accagctcga gggtgagtca cgaaagtcat gtagcctgca ccaggctccc accgtccgcg
      901 gctccgtgcg tttctggtgt tcactgccaa cagcggacac atcgagtgct gtgccgctgg
      961 agctgcaggt gacggaggcg tccggttctc ctcgctatca ccgcatcatc catatcaatg
     1021 aagtagtgct cctggacgcc cccgcggggc tgctggcgcg ccgggcagaa gagggcagcc
     1081 acgtggtgct gcgctggctg ccacctcctg gagcacctat gaccacccac atccgatatg
     1141 aagtggacgt gtcggcaggc aaccgggcag gagggacaca aagggtggag gtcctggaag
     1201 gccgcactga gtgtgttctg agcaacctgc ggggcgggac gcgctacacc ttcgctgttc
     1261 gagcgcgcat ggccgagccg agcttcagcg gattctggag tgcctggtct gagcccgcgt
     1321 cactactgac cgctagcgac ctggaccctc tcatcttgac gctgtctctc attctggtcc
     1381 tcatctcgtt gttgctgacg gttctggccc tgctgtccat gaagagcggc accaagaaga
     1441 gcgacttcca tagccagatg gctgtgcaca agctggccaa gagcatccct ctgcgcagac
     1501 aggtaacagt gtcagctgac tccagtgcat ccatgaactc tggggttctc ctggttcggc
     1561 cctcacggct ctcctccagc gggaccccca tgctggctgg agtctccgaa tatgagctcc
     1621 ctgaggatcc ccgctgggag ctgccacgag acagactggt cttaggcaaa ccacttggcg
     1681 agggctgctt cgggcaggtg gtgttggctg aggccatcgg gctggataag gacaaaccca
     1741 accgtgtgac caaagtggcc gtgaagatgt tgaagtccga cgcaacggag aaggacctgt
     1801 cggatctgat ctcggagatg gagatgatga aaatgattgg gaagcacaag aatatcatca
     1861 accttctggg agcgtgcaca caggatggtc ctctttatgt cattgtggag tacgcctcca
     1921 aaggcaatct ccgggagtat ctacaggccc ggaggcctcc tgggctggag tactgctata
     1981 accccagcca caaccccgag gaacagctgt cttccaaaga tctggtatcc tgtgcctatc
     2041 aggtggctcg gggcatggag tatcttgcct ctaagaagtg tatacaccga gacctggctg
     2101 ctaggaacgt cctggtgacc gaggataacg taatgaagat cgcagacttt ggcttagctc
     2161 gagacattca tcatatcgac tactacaaga aaaccaccaa cggccggctg cctgtgaagt
     2221 ggatggcccc tgaggcgttg tttgaccgga tctacacaca ccagagcgat gtgtggtctt
     2281 ttggagtgct cttgtgggag atcttcactc tgggtggctc cccatacccc ggtgtgcctg
     2341 tggaggaact tttcaagctg ctgaaggagg gtcatcgaat ggacaagccc agtaactgta
     2401 ccaatgagct gtacatgatg atgcgggact gctggcatgc agtgccctct cagagaccta
     2461 cgttcaagca gttggtggaa gacctggacc gcattgtggc cttgacctcc aaccaggagt
     2521 atctggacct gtccataccg ctggaccagt actcacccag ctttcccgac acacggagct
     2581 ccacctgctc ctcaggggag gactctgtct tctctcatga gccgttacct gaggagccct
     2641 gtctgcctcg acaccccacc cagcttgcca acagtggact caaacggcgc taggcggccg
     2701 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     2761 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2821 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2881 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2941 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3001 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3061 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3121 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3181 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3241 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3301 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3361 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3421 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3481 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3541 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3601 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3661 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3721 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3781 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     3841 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3901 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3961 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4021 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4081 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4141 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4201 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4261 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4321 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4381 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4441 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4501 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4561 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4621 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4681 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4741 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4801 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4861 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4921 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     4981 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5041 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5101 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5161 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5221 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5281 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5341 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5401 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5461 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5521 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5581 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5641 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5701 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5761 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5821 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5881 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg
     5941 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6001 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6061 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6121 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6181 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6241 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6301 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6361 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6421 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6481 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6541 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6601 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6661 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6721 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6781 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6841 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6901 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6961 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7021 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7081 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7141 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     7201 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     7261 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     7321 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata t
//
