LOCUS       40924_28062        6924 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo627, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6924)
  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 6924)
  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 6924)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6924)
  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..6924
                     /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"
     sig_peptide     246..308
                     /label=Ig-kappa leader
                     /note="leader sequence from mouse immunoglobulin kappa 
                     light
                     chain"
     misc_feature    310..654
                     /label=VL
                     /note="VL"
     misc_feature    660..1403
                     /label=EpoR-D1_D2_TM
                     /note="EpoR-D1_D2_TM"
     misc_feature    933..935
                     /label=F93A
                     /note="F93A"
     misc_feature    1335..1403
                     /label=transmembrane
                     /note="transmembrane"
     misc_feature    1413..2241
                     /label=IL6ST
                     /note="IL6ST"
     misc_feature    1764..1766
                     /label=Y759A
                     /note="Y759A"
     polyA_signal    2297..2521
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2567..2995
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3009..3338
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3405..4196
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4373..4506
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4543..4559)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4567..4583)
                     /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(4591..4621)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4636..4657)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4945..5530)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5704..6561)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6562..6666)
                     /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 tccgcagcag caaataagcg agacctaatt aaaaaacaca
     1441 tctggcctaa tgttccagat ccttcaaaga gtcatattgc ccagtggtca cctcacactc
     1501 ctccaaggca caattttaat tcaaaagatc aaatgtattc agatggcaat ttcactgatg
     1561 taagtgttgt ggaaatagaa gcaaatgaca aaaagccttt tccagaagat ctgaaatcat
     1621 tggacctgtt caaaaaggaa aaaattaata ctgaaggaca cagcagtggt attggggggt
     1681 cttcatgcat gtcatcttct aggccaagca tttctagcag tgatgaaaat gaatcttcac
     1741 aaaacacttc gagcactgtc caggcttcta ccgtggtaca cagtggctac agacaccaag
     1801 ttccgtcagt ccaagtcttc tcaagatccg agtctaccca gcccttgtta gattcagagg
     1861 agcggccaga agatctacaa ttagtagatc atgtagatgg cggtgatggt attttgccca
     1921 ggcaacagta cttcaaacag aactgcagtc agcatgaatc cagtccagat atttcacatt
     1981 ttgaaaggtc aaagcaagtt tcatcagtca atgaggaaga ttttgttaga cttaaacagc
     2041 agatttcaga tcatatttca caatcctgtg gatctgggca aatgaaaatg tttcaggaag
     2101 tttctgcagc agatgctttt ggtccaggta ctgagggaca agtagaaaga tttgaaacag
     2161 ttggcatgga ggctgcgact gatgaaggca tgcctaaaag ttacttacca cagactgtac
     2221 ggcaaggcgg ctacatgcct cagtaggcgg ccgctcgagt ctagagggcc cgtttaaacc
     2281 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2341 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2401 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2461 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2521 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     2581 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2641 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2701 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2761 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2821 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2881 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2941 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3001 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3061 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3121 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3181 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3241 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3301 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     3361 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3421 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3481 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3541 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3601 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3661 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3721 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3781 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3841 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3901 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     3961 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4021 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4081 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4141 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4201 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4261 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4321 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4381 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4441 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4501 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     4561 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4621 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4681 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4741 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4801 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4861 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4921 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4981 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5041 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5101 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5161 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5221 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5281 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5341 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5401 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5461 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5521 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5581 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5641 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5701 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5761 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5821 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5881 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5941 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6001 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6061 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6121 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6181 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6241 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6301 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6361 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6421 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6481 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6541 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6601 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6661 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     6721 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     6781 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     6841 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     6901 cttcgcgatg tacgggccag atat
//
