LOCUS       40924_28122        7847 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo690, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7847)
  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 7847)
  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 7847)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7847)
  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..7847
                     /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..692
                     /label=anti RR120
                     /note="anti RR120"
     misc_feature    705..1448
                     /label=EpoR-D1_D2_TM
                     /note="EpoR-D1_D2_TM"
     misc_feature    978..980
                     /label=F93A
                     /note="F93A"
     misc_feature    1380..1448
                     /label=transmembrane
                     /note="transmembrane"
     misc_feature    1449..3161
                     /label=VEGFR
                     /note="VEGFR"
     polyA_signal    3220..3444
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3490..3918
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3932..4261
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4328..5119
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    5296..5429
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5466..5482)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5490..5506)
                     /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(5514..5544)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5559..5580)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5868..6453)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6627..7484)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7485..7589)
                     /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 tgctatcact acggaccgtt aagcgggcca atggagggga actgaagaca ggctacttgt
     1501 ccatcgtcat ggatccagat gaactcccat tggatgaaca ttgtgaacga ctgccttatg
     1561 atgccagcaa atgggaattc cccagagacc ggctgaagct aggtaagcct cttggccgtg
     1621 gtgcctttgg ccaagtgatt gaagcagatg cctttggaat tgacaagaca gcaacttgca
     1681 ggacagtagc agtcaaaatg ttgaaagaag gagcaacaca cagtgagcat cgagctctca
     1741 tgtctgaact caagatcctc attcatattg gtcaccatct caatgtggtc aaccttctag
     1801 gtgcctgtac caagccagga gggccactca tggtgattgt ggaattctgc aaatttggaa
     1861 acctgtccac ttacctgagg agcaagagaa atgaatttgt cccctacaag accaaagggg
     1921 cacgattccg tcaagggaaa gactacgttg gagcaatccc tgtggatctg aaacggcgct
     1981 tggacagcat caccagtagc cagagctcag ccagctctgg atttgtggag gagaagtccc
     2041 tcagtgatgt agaagaagag gaagctcctg aagatctgta taaggacttc ctgaccttgg
     2101 agcatctcat ctgttacagc ttccaagtgg ctaagggcat ggagttcttg gcatcgcgaa
     2161 agtgtatcca cagggacctg gcggcacgaa atatcctctt atcggagaag aacgtggtta
     2221 aaatctgtga ctttggcttg gcccgggata tttataaaga tccagattat gtcagaaaag
     2281 gagatgctcg cctccctttg aaatggatgg ccccagaaac aatttttgac agagtgtaca
     2341 caatccagag tgacgtctgg tcttttggtg ttttgctgtg ggaaatattt tccttaggtg
     2401 cttctccata tcctggggta aagattgatg aagaattttg taggcgattg aaagaaggaa
     2461 ctagaatgag ggcccctgat tatactacac cagaaatgta ccagaccatg ctggactgct
     2521 ggcacgggga gcccagtcag agacccacgt tttcagagtt ggtggaacat ttgggaaatc
     2581 tcttgcaagc taatgctcag caggatggca aagactacat tgttcttccg atatcagaga
     2641 ctttgagcat ggaagaggat tctggactct ctctgcctac ctcacctgtt tcctgtatgg
     2701 aggaggagga agtatgtgac cccaaattcc attatgacaa cacagcagga atcagtcagt
     2761 atctgcagaa cagtaagcga aagagccggc ctgtgagtgt aaaaacattt gaagatatcc
     2821 cgttagaaga accagaagta aaagtaatcc cagatgacaa ccagacggac agtggtatgg
     2881 ttcttgcctc agaagagctg aaaactttgg aagacagaac caaattatct ccatcttttg
     2941 gtggaatggt gcccagcaaa agcagggagt ctgtggcatc tgaaggctca aaccagacaa
     3001 gcggctacca gtccggatat cactccgatg acacagacac caccgtgtac tccagtgagg
     3061 aagcagaact tttaaagctg atagagattg gagtgcaaac cggtagcaca gcccagattc
     3121 tccagcctga ctcggggacc acactgagct ctcctcctgt tacgcgtacg cggccgctcg
     3181 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     3241 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3301 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3361 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3421 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     3481 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3541 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3601 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3661 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3721 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3781 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3841 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3901 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3961 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4021 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4081 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4141 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4201 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4261 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     4321 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4381 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4441 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4501 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4561 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4621 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4681 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4741 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4801 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     4861 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4921 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4981 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5041 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5101 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     5161 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5221 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5281 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5341 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5401 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     5461 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5521 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5581 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5641 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5701 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5761 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5821 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5881 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5941 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6001 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6061 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6121 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6181 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6241 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6301 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6361 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag
     6421 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6481 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6541 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6601 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6661 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6721 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6781 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6841 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6901 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6961 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7021 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7081 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7141 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7201 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7261 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7321 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7381 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7441 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7501 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7561 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     7621 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
     7681 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     7741 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     7801 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatat
//
