LOCUS       40924_28082        7410 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo644, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7410)
  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 7410)
  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 7410)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7410)
  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..7410
                     /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    246..294
                     /label=secrete
                     /note="secrete"
     misc_feature    309
                     /label=scfv
                     /note="scfv"
     misc_feature    315..692
                     /label=anti RR120
                     /note="anti RR120"
     misc_feature    705..1448
                     /label=mouse EpoR-D1_D2_TM
                     /note="mouse EpoR-D1_D2_TM"
     misc_feature    978..980
                     /label=F93A
                     /note="F93A"
     misc_feature    1380..1448
                     /label=transmembrane
                     /note="transmembrane"
     misc_feature    1458..2729
                     /label=mFGFR 405-end
                     /note="mFGFR 405-end"
     polyA_signal    2783..3007
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3053..3481
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3495..3824
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3891..4682
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    4859..4992
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5029..5045)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5053..5069)
                     /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(5077..5107)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5122..5143)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5431..6016)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6190..7047)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7048..7152)
                     /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 atccgtgcag ttgcaagaat cgggtggtgg actggttcaa gccggtgata
      361 gtttgaaact gtcatgcgaa gccagtggcg attccattgg aacttatgtc attggttggt
      421 ttcgccaagc accggggaaa gagcgcattt accttgccac tattggtcgt aacctggtcg
      481 ggccgtccga tttttataca cgctacgctg actctgtaaa gggtcgcttc gcagtgagtc
      541 gtgacaacgc caagaacact gtaaacttac agatgaactc tcttaaacct gaagatactg
      601 ctgtttacta ttgtgctgcg aaaacgacaa cgtggggggg aaacgatcct aataactgga
      661 attattgggg acaaggcact caggtgaccg tatccggaga attcgcacct tcacccagcc
      721 tcccggaccc caagtttgag agcaaagcgg ccctgctggc atcccggggc tccgaagaac
      781 ttctgtgctt cacccaacgc ttggaagact tggtgtgttt ctgggaggaa gcggcgagct
      841 ccgggatgga cttcaactac agcttctcat accagctcga gggtgagtca cgaaagtcat
      901 gtagcctgca ccaggctccc accgtccgcg gctccgtgcg tttctggtgt tcactgccaa
      961 cagcggacac atcgagtgct gtgccgctgg agctgcaggt gacggaggcg tccggttctc
     1021 ctcgctatca ccgcatcatc catatcaatg aagtagtgct cctggacgcc cccgcggggc
     1081 tgctggcgcg ccgggcagaa gagggcagcc acgtggtgct gcgctggctg ccacctcctg
     1141 gagcacctat gaccacccac atccgatatg aagtggacgt gtcggcaggc aaccgggcag
     1201 gagggacaca aagggtggag gtcctggaag gccgcactga gtgtgttctg agcaacctgc
     1261 ggggcgggac gcgctacacc ttcgctgttc gagcgcgcat ggccgagccg agcttcagcg
     1321 gattctggag tgcctggtct gagcccgcgt cactactgac cgctagcgac ctggaccctc
     1381 tcatcttgac gctgtctctc attctggtcc tcatctcgtt gttgctgacg gttctggccc
     1441 tgctgtccgc cgctgcgatg aagagcggca ccaagaagag cgacttccat agccagatgg
     1501 ctgtgcacaa gctggccaag agcatccctc tgcgcagaca ggtaacagtg tcagctgact
     1561 ccagtgcatc catgaactct ggggttctcc tggttcggcc ctcacggctc tcctccagcg
     1621 ggacccccat gctggctgga gtctccgaat atgagctccc tgaggatccc cgctgggagc
     1681 tgccacgaga cagactggtc ttaggcaaac cacttggcga gggctgcttc gggcaggtgg
     1741 tgttggctga ggccatcggg ctggataagg acaaacccaa ccgtgtgacc aaagtggccg
     1801 tgaagatgtt gaagtccgac gcaacggaga aggacctgtc ggatctgatc tcggagatgg
     1861 agatgatgaa aatgattggg aagcacaaga atatcatcaa ccttctggga gcgtgcacac
     1921 aggatggtcc tctttatgtc attgtggagt acgcctccaa aggcaatctc cgggagtatc
     1981 tacaggcccg gaggcctcct gggctggagt actgctataa ccccagccac aaccccgagg
     2041 aacagctgtc ttccaaagat ctggtatcct gtgcctatca ggtggctcgg ggcatggagt
     2101 atcttgcctc taagaagtgt atacaccgag acctggctgc taggaacgtc ctggtgaccg
     2161 aggataacgt aatgaagatc gcagactttg gcttagctcg agacattcat catatcgact
     2221 actacaagaa aaccaccaac ggccggctgc ctgtgaagtg gatggcccct gaggcgttgt
     2281 ttgaccggat ctacacacac cagagcgatg tgtggtcttt tggagtgctc ttgtgggaga
     2341 tcttcactct gggtggctcc ccataccccg gtgtgcctgt ggaggaactt ttcaagctgc
     2401 tgaaggaggg tcatcgaatg gacaagccca gtaactgtac caatgagctg tacatgatga
     2461 tgcgggactg ctggcatgca gtgccctctc agagacctac gttcaagcag ttggtggaag
     2521 acctggaccg cattgtggcc ttgacctcca accaggagta tctggacctg tccataccgc
     2581 tggaccagta ctcacccagc tttcccgaca cacggagctc cacctgctcc tcaggggagg
     2641 actctgtctt ctctcatgag ccgttacctg aggagccctg tctgcctcga caccccaccc
     2701 agcttgccaa cagtggactc aaacggcgct aggcggccgc tcgagtctag agggcccgtt
     2761 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2821 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2881 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2941 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     3001 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     3061 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3121 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3181 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3241 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3301 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3361 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3421 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3481 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3541 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3601 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3661 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3721 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3781 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3841 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3901 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3961 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4021 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     4081 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4141 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4201 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4261 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4321 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4381 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4441 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4501 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4561 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4621 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4681 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4741 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4801 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4861 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4921 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4981 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5041 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5101 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5161 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5221 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5281 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5341 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5401 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5461 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5521 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5581 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5641 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5701 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5761 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5821 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5881 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5941 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     6001 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6061 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6121 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6181 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6241 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6301 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6361 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6421 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6481 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6541 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6601 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6661 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6721 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6781 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6841 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6901 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6961 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7021 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7081 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7141 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     7201 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     7261 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     7321 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
     7381 gcgctgcttc gcgatgtacg ggccagatat
//
