LOCUS       40924_28077        7407 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo643, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7407)
  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 7407)
  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 7407)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7407)
  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..7407
                     /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    1455..2726
                     /label=mFGFR 405-end
                     /note="mFGFR 405-end"
     polyA_signal    2780..3004
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3050..3478
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3492..3821
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3888..4679
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    4856..4989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5026..5042)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5050..5066)
                     /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(5074..5104)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5119..5140)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5428..6013)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6187..7044)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7045..7149)
                     /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 tgcgatgaag agcggcacca agaagagcga cttccatagc cagatggctg
     1501 tgcacaagct ggccaagagc atccctctgc gcagacaggt aacagtgtca gctgactcca
     1561 gtgcatccat gaactctggg gttctcctgg ttcggccctc acggctctcc tccagcggga
     1621 cccccatgct ggctggagtc tccgaatatg agctccctga ggatccccgc tgggagctgc
     1681 cacgagacag actggtctta ggcaaaccac ttggcgaggg ctgcttcggg caggtggtgt
     1741 tggctgaggc catcgggctg gataaggaca aacccaaccg tgtgaccaaa gtggccgtga
     1801 agatgttgaa gtccgacgca acggagaagg acctgtcgga tctgatctcg gagatggaga
     1861 tgatgaaaat gattgggaag cacaagaata tcatcaacct tctgggagcg tgcacacagg
     1921 atggtcctct ttatgtcatt gtggagtacg cctccaaagg caatctccgg gagtatctac
     1981 aggcccggag gcctcctggg ctggagtact gctataaccc cagccacaac cccgaggaac
     2041 agctgtcttc caaagatctg gtatcctgtg cctatcaggt ggctcggggc atggagtatc
     2101 ttgcctctaa gaagtgtata caccgagacc tggctgctag gaacgtcctg gtgaccgagg
     2161 ataacgtaat gaagatcgca gactttggct tagctcgaga cattcatcat atcgactact
     2221 acaagaaaac caccaacggc cggctgcctg tgaagtggat ggcccctgag gcgttgtttg
     2281 accggatcta cacacaccag agcgatgtgt ggtcttttgg agtgctcttg tgggagatct
     2341 tcactctggg tggctcccca taccccggtg tgcctgtgga ggaacttttc aagctgctga
     2401 aggagggtca tcgaatggac aagcccagta actgtaccaa tgagctgtac atgatgatgc
     2461 gggactgctg gcatgcagtg ccctctcaga gacctacgtt caagcagttg gtggaagacc
     2521 tggaccgcat tgtggccttg acctccaacc aggagtatct ggacctgtcc ataccgctgg
     2581 accagtactc acccagcttt cccgacacac ggagctccac ctgctcctca ggggaggact
     2641 ctgtcttctc tcatgagccg ttacctgagg agccctgtct gcctcgacac cccacccagc
     2701 ttgccaacag tggactcaaa cggcgctagg cggccgctcg agtctagagg gcccgtttaa
     2761 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     2821 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     2881 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     2941 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     3001 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     3061 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3121 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3181 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3241 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3301 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3361 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3421 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     3481 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3541 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3601 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3661 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3721 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     3781 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     3841 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3901 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3961 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4021 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     4081 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4141 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4201 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4261 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4321 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4381 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     4441 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4501 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4561 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4621 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4681 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4741 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4801 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4861 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4921 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4981 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5041 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5101 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5161 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5221 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5281 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5341 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5401 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5461 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5521 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5581 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5641 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5701 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5761 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5821 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5881 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5941 caaacaaacc accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6001 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6061 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6121 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6181 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6241 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6301 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6361 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6421 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6481 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6541 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6601 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6661 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6721 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6781 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6841 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6901 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6961 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7021 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7081 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7141 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     7201 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     7261 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     7321 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
     7381 ctgcttcgcg atgtacgggc cagatat
//
