LOCUS       40924_28102        7356 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo668, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7356)
  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 7356)
  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 7356)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7356)
  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..7356
                     /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..654
                     /label=VL
                     /note="VL"
     misc_feature    660..1403
                     /label=mouse EpoR-D1_D2_TM
                     /note="mouse EpoR-D1_D2_TM"
     misc_feature    933..935
                     /label=F93A
                     /note="F93A"
     misc_feature    1335..1403
                     /label=transmembrane
                     /note="transmembrane"
     misc_feature    1404..2675
                     /label=mFGFR 405-end
                     /note="mFGFR 405-end"
     polyA_signal    2729..2953
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2999..3427
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3441..3770
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3837..4628
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    4805..4938
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4975..4991)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4999..5015)
                     /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(5023..5053)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5068..5089)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5377..5962)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6136..6993)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6994..7098)
                     /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 atcccaatcc gagttgaccc agccgcctag tgctagtgga acaccgggac
      361 agcgcgtgac aatttcatgc tccggttcaa gctcaaatat cggctctaat tatgtttact
      421 ggtatcagca gcttccaggt actgcgccta agctcttaat ctaccgtaac aatcaacgtc
      481 cgtccggtgt gcccgaccgc ttctcaggca gtaaaagtgg tacttccgca tcccttgcaa
      541 tttcgggact tcgtagcgaa gatgaggcag actattactg tgcagcatgg gatgattcct
      601 tatcagcttg ggtattcggt ggcggaactc aattagacat tttggggtcc ggagaattcg
      661 caccttcacc cagcctcccg gaccccaagt ttgagagcaa agcggccctg ctggcatccc
      721 ggggctccga agaacttctg tgcttcaccc aacgcttgga agacttggtg tgtttctggg
      781 aggaagcggc gagctccggg atggacttca actacagctt ctcataccag ctcgagggtg
      841 agtcacgaaa gtcatgtagc ctgcaccagg ctcccaccgt ccgcggctcc gtgcgtttct
      901 ggtgttcact gccaacagcg gacacatcga gtgctgtgcc gctggagctg caggtgacgg
      961 aggcgtccgg ttctcctcgc tatcaccgca tcatccatat caatgaagta gtgctcctgg
     1021 acgcccccgc ggggctgctg gcgcgccggg cagaagaggg cagccacgtg gtgctgcgct
     1081 ggctgccacc tcctggagca cctatgacca cccacatccg atatgaagtg gacgtgtcgg
     1141 caggcaaccg ggcaggaggg acacaaaggg tggaggtcct ggaaggccgc actgagtgtg
     1201 ttctgagcaa cctgcggggc gggacgcgct acaccttcgc tgttcgagcg cgcatggccg
     1261 agccgagctt cagcggattc tggagtgcct ggtctgagcc cgcgtcacta ctgaccgcta
     1321 gcgacctgga ccctctcatc ttgacgctgt ctctcattct ggtcctcatc tcgttgttgc
     1381 tgacggttct ggccctgctg tccatgaaga gcggcaccaa gaagagcgac ttccatagcc
     1441 agatggctgt gcacaagctg gccaagagca tccctctgcg cagacaggta acagtgtcag
     1501 ctgactccag tgcatccatg aactctgggg ttctcctggt tcggccctca cggctctcct
     1561 ccagcgggac ccccatgctg gctggagtct ccgaatatga gctccctgag gatccccgct
     1621 gggagctgcc acgagacaga ctggtcttag gcaaaccact tggcgagggc tgcttcgggc
     1681 aggtggtgtt ggctgaggcc atcgggctgg ataaggacaa acccaaccgt gtgaccaaag
     1741 tggccgtgaa gatgttgaag tccgacgcaa cggagaagga cctgtcggat ctgatctcgg
     1801 agatggagat gatgaaaatg attgggaagc acaagaatat catcaacctt ctgggagcgt
     1861 gcacacagga tggtcctctt tatgtcattg tggagtacgc ctccaaaggc aatctccggg
     1921 agtatctaca ggcccggagg cctcctgggc tggagtactg ctataacccc agccacaacc
     1981 ccgaggaaca gctgtcttcc aaagatctgg tatcctgtgc ctatcaggtg gctcggggca
     2041 tggagtatct tgcctctaag aagtgtatac accgagacct ggctgctagg aacgtcctgg
     2101 tgaccgagga taacgtaatg aagatcgcag actttggctt agctcgagac attcatcata
     2161 tcgactacta caagaaaacc accaacggcc ggctgcctgt gaagtggatg gcccctgagg
     2221 cgttgtttga ccggatctac acacaccaga gcgatgtgtg gtcttttgga gtgctcttgt
     2281 gggagatctt cactctgggt ggctccccat accccggtgt gcctgtggag gaacttttca
     2341 agctgctgaa ggagggtcat cgaatggaca agcccagtaa ctgtaccaat gagctgtaca
     2401 tgatgatgcg ggactgctgg catgcagtgc cctctcagag acctacgttc aagcagttgg
     2461 tggaagacct ggaccgcatt gtggccttga cctccaacca ggagtatctg gacctgtcca
     2521 taccgctgga ccagtactca cccagctttc ccgacacacg gagctccacc tgctcctcag
     2581 gggaggactc tgtcttctct catgagccgt tacctgagga gccctgtctg cctcgacacc
     2641 ccacccagct tgccaacagt ggactcaaac ggcgctaggc ggccgctcga gtctagaggg
     2701 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2761 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2821 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2881 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     2941 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     3001 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3061 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3121 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3181 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3241 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3301 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3361 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3421 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3481 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3541 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3601 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3661 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3721 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3781 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3841 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3901 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3961 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     4021 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4081 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4141 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4201 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4261 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4321 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     4381 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4441 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4501 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4561 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4621 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4681 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4741 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4801 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4861 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4921 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4981 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5041 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5101 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5161 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5221 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5281 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5341 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5401 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5461 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5521 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5581 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5641 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5701 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5761 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5821 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5881 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac
     5941 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6001 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6061 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6121 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6181 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6241 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6301 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6361 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6421 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6481 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6541 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6601 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6661 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6721 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6781 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6841 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6901 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6961 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7021 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7081 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     7141 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     7201 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     7261 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
     7321 cgttttgcgc tgcttcgcga tgtacgggcc agatat
//
