LOCUS       40924_28117        7770 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo671, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7770)
  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 7770)
  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 7770)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7770)
  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..7770
                     /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    310..314
                     /label=heavy
                     /note="heavy"
     misc_feature    315..650
                     /label=light
                     /note="light"
     misc_feature    711..1061
                     /label=heavy
                     /note="heavy"
     misc_feature    1074..1817
                     /label=mouse EpoR-D1_D2_TM
                     /note="mouse EpoR-D1_D2_TM"
     misc_feature    1347..1349
                     /label=F93A
                     /note="F93A"
     misc_feature    1749..1817
                     /label=transmembrane
                     /note="transmembrane"
     misc_feature    1818..3089
                     /label=mFGFR 405-end
                     /note="mFGFR 405-end"
     polyA_signal    3143..3367
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3413..3841
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3855..4184
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4251..5042
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    5219..5352
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5389..5405)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5413..5429)
                     /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(5437..5467)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5482..5503)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5791..6376)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6550..7407)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7408..7512)
                     /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 atccgacatc gttcttacgc aatctcctcc gtccttggct gtgagcctgg
      361 gtcaacgtgc caccatctca tgtcgtgcga gcgagagtat tgatttgtac ggattcacat
      421 ttatgcactg gtatcagcaa aagcccggtc aaccacccaa aattctgatt taccgcgcgt
      481 caaatttaga gtcaggcatt cctgcgcgct tctcaggttc gggctcacgc acagacttta
      541 cattgacaat taacccagtg gaagcagacg atgttgcgac atattattgt caacagacgc
      601 acgaagatcc atacacattt ggtggcggca ctaaattgga aattaagcgt ggtggtggag
      661 gatcaggtgg tggagggtct ggcggaggcg gtagcggtgg aggtggaagc caagtacagt
      721 tacagcagag tggcgcagag ttggcaaaac cgggagcctc ggttaagatg tcatgcaaga
      781 cgagtggata ctcatttagc agttactgga tgcactgggt taaacaacgc ccagggcaag
      841 gacttgaatg gattgggtat atcaatccat cgaccggata tacagagaac aaccagaagt
      901 ttaaagacaa ggttactctg accgctgaca agtcaagtaa cacggcatat atgcaattaa
      961 acagccttac ctctgaagat tcagccgtct attactgtgc tcgctcaggg cgtctgtact
     1021 tcgatgtatg gggggcaggt acgacagtga cggtgtctag ttccggagaa ttcgcacctt
     1081 cacccagcct cccggacccc aagtttgaga gcaaagcggc cctgctggca tcccggggct
     1141 ccgaagaact tctgtgcttc acccaacgct tggaagactt ggtgtgtttc tgggaggaag
     1201 cggcgagctc cgggatggac ttcaactaca gcttctcata ccagctcgag ggtgagtcac
     1261 gaaagtcatg tagcctgcac caggctccca ccgtccgcgg ctccgtgcgt ttctggtgtt
     1321 cactgccaac agcggacaca tcgagtgctg tgccgctgga gctgcaggtg acggaggcgt
     1381 ccggttctcc tcgctatcac cgcatcatcc atatcaatga agtagtgctc ctggacgccc
     1441 ccgcggggct gctggcgcgc cgggcagaag agggcagcca cgtggtgctg cgctggctgc
     1501 cacctcctgg agcacctatg accacccaca tccgatatga agtggacgtg tcggcaggca
     1561 accgggcagg agggacacaa agggtggagg tcctggaagg ccgcactgag tgtgttctga
     1621 gcaacctgcg gggcgggacg cgctacacct tcgctgttcg agcgcgcatg gccgagccga
     1681 gcttcagcgg attctggagt gcctggtctg agcccgcgtc actactgacc gctagcgacc
     1741 tggaccctct catcttgacg ctgtctctca ttctggtcct catctcgttg ttgctgacgg
     1801 ttctggccct gctgtccatg aagagcggca ccaagaagag cgacttccat agccagatgg
     1861 ctgtgcacaa gctggccaag agcatccctc tgcgcagaca ggtaacagtg tcagctgact
     1921 ccagtgcatc catgaactct ggggttctcc tggttcggcc ctcacggctc tcctccagcg
     1981 ggacccccat gctggctgga gtctccgaat atgagctccc tgaggatccc cgctgggagc
     2041 tgccacgaga cagactggtc ttaggcaaac cacttggcga gggctgcttc gggcaggtgg
     2101 tgttggctga ggccatcggg ctggataagg acaaacccaa ccgtgtgacc aaagtggccg
     2161 tgaagatgtt gaagtccgac gcaacggaga aggacctgtc ggatctgatc tcggagatgg
     2221 agatgatgaa aatgattggg aagcacaaga atatcatcaa ccttctggga gcgtgcacac
     2281 aggatggtcc tctttatgtc attgtggagt acgcctccaa aggcaatctc cgggagtatc
     2341 tacaggcccg gaggcctcct gggctggagt actgctataa ccccagccac aaccccgagg
     2401 aacagctgtc ttccaaagat ctggtatcct gtgcctatca ggtggctcgg ggcatggagt
     2461 atcttgcctc taagaagtgt atacaccgag acctggctgc taggaacgtc ctggtgaccg
     2521 aggataacgt aatgaagatc gcagactttg gcttagctcg agacattcat catatcgact
     2581 actacaagaa aaccaccaac ggccggctgc ctgtgaagtg gatggcccct gaggcgttgt
     2641 ttgaccggat ctacacacac cagagcgatg tgtggtcttt tggagtgctc ttgtgggaga
     2701 tcttcactct gggtggctcc ccataccccg gtgtgcctgt ggaggaactt ttcaagctgc
     2761 tgaaggaggg tcatcgaatg gacaagccca gtaactgtac caatgagctg tacatgatga
     2821 tgcgggactg ctggcatgca gtgccctctc agagacctac gttcaagcag ttggtggaag
     2881 acctggaccg cattgtggcc ttgacctcca accaggagta tctggacctg tccataccgc
     2941 tggaccagta ctcacccagc tttcccgaca cacggagctc cacctgctcc tcaggggagg
     3001 actctgtctt ctctcatgag ccgttacctg aggagccctg tctgcctcga caccccaccc
     3061 agcttgccaa cagtggactc aaacggcgct aggcggccgc tcgagtctag agggcccgtt
     3121 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     3181 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3241 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3301 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     3361 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     3421 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3481 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3541 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3601 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3661 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3721 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3781 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3841 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3901 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3961 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4021 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4081 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     4141 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     4201 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4261 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4321 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4381 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     4441 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4501 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4561 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4621 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4681 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4741 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4801 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4861 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4921 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4981 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5041 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5101 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5161 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     5221 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5281 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5341 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5401 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5461 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5521 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5581 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5641 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5701 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5761 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5821 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5881 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5941 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6001 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6061 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6121 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6181 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6241 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6301 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     6361 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6421 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6481 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6541 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6601 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6661 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6721 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6781 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6841 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6901 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6961 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7021 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7081 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7141 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7201 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7261 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7321 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7381 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7441 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7501 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     7561 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     7621 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     7681 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
     7741 gcgctgcttc gcgatgtacg ggccagatat
//
