LOCUS       40924_28042        7344 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo621, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7344)
  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 7344)
  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 7344)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7344)
  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..7344
                     /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    315..1068
                     /label=5d3d11
                     /note="5d3d11"
     misc_feature    315..650
                     /label=light
                     /note="light"
     misc_feature    714..1068
                     /label=heavy
                     /note="heavy"
     misc_feature    1080..1823
                     /label=EpoR-D1_D2_TM
                     /note="EpoR-D1_D2_TM"
     misc_feature    1353..1355
                     /label=F93A
                     /note="F93A"
     misc_feature    1755..1823
                     /label=transmembrane
                     /note="transmembrane"
     misc_feature    1833..2661
                     /label=IL6ST
                     /note="IL6ST"
     misc_feature    2184..2186
                     /label=Y759A
                     /note="Y759A"
     polyA_signal    2717..2941
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2987..3415
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3429..3758
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3825..4616
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4793..4926
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4963..4979)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4987..5003)
                     /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(5011..5041)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5056..5077)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5365..5950)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6124..6981)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6982..7086)
                     /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 atccgacatt gttatgacgc agaccgcgcc atccgtcttt gtgacgcccg
      361 gtgaatccgt cagtatctca tgtcgttctt ctaagagttt attgcatagc aatggaaata
      421 catatttata ctggttcctt caacgtccgg gccaaagccc ccagttactt atttatcgta
      481 tgtccaatct tgcatcaggt gtccccgacc gtttcagtgg atctgggtcc ggcacggact
      541 tcactcttcg tatttcgcgt gtagaagccg aagatgtagg agtctattac tgtatgcaac
      601 atctggaata cccagtcacc tttggcgccg gtactaaggt ggaaatcaag cgtggtggtg
      661 gaggatcagg tggtggaggg tctggcggag gcggtagcgg tggaggtgga agccaagtac
      721 agttacaaca atccgggccg gagttagtaa aacccggtgc atcggtcaag atcagttgca
      781 aggtgtcggg ttatgcgatt tcatcttcct ggatgaactg ggttaaacaa cgtcctggac
      841 acggcttaga atggattggt cgtatttacc caggggatgg ggatacaaaa tataacggga
      901 aattcaagga taaggccaca cttacggtag acaaaagctc ctcaactgct tacatgcagc
      961 tttcctcgct tacgagcgtt gattcggctg tgtacttttg tgcccgcgat ggctaccgct
     1021 attattttga ttattggggg caaggtacta gcgtgacggt tagctcatcc ggagaattcg
     1081 caccttcacc cagcctcccg gaccccaagt ttgagagcaa agcggccctg ctggcatccc
     1141 ggggctccga agaacttctg tgcttcaccc aacgcttgga agacttggtg tgtttctggg
     1201 aggaagcggc gagctccggg atggacttca actacagctt ctcataccag ctcgagggtg
     1261 agtcacgaaa gtcatgtagc ctgcaccagg ctcccaccgt ccgcggctcc gtgcgtttct
     1321 ggtgttcact gccaacagcg gacacatcga gtgctgtgcc gctggagctg caggtgacgg
     1381 aggcgtccgg ttctcctcgc tatcaccgca tcatccatat caatgaagta gtgctcctgg
     1441 acgcccccgc ggggctgctg gcgcgccggg cagaagaggg cagccacgtg gtgctgcgct
     1501 ggctgccacc tcctggagca cctatgacca cccacatccg atatgaagtg gacgtgtcgg
     1561 caggcaaccg ggcaggaggg acacaaaggg tggaggtcct ggaaggccgc actgagtgtg
     1621 ttctgagcaa cctgcggggc gggacgcgct acaccttcgc tgttcgagcg cgcatggccg
     1681 agccgagctt cagcggattc tggagtgcct ggtctgagcc cgcgtcacta ctgaccgcta
     1741 gcgacctgga ccctctcatc ttgacgctgt ctctcattct ggtcctcatc tcgttgttgc
     1801 tgacggttct ggccctgctg tccgcagcag caaataagcg agacctaatt aaaaaacaca
     1861 tctggcctaa tgttccagat ccttcaaaga gtcatattgc ccagtggtca cctcacactc
     1921 ctccaaggca caattttaat tcaaaagatc aaatgtattc agatggcaat ttcactgatg
     1981 taagtgttgt ggaaatagaa gcaaatgaca aaaagccttt tccagaagat ctgaaatcat
     2041 tggacctgtt caaaaaggaa aaaattaata ctgaaggaca cagcagtggt attggggggt
     2101 cttcatgcat gtcatcttct aggccaagca tttctagcag tgatgaaaat gaatcttcac
     2161 aaaacacttc gagcactgtc caggcttcta ccgtggtaca cagtggctac agacaccaag
     2221 ttccgtcagt ccaagtcttc tcaagatccg agtctaccca gcccttgtta gattcagagg
     2281 agcggccaga agatctacaa ttagtagatc atgtagatgg cggtgatggt attttgccca
     2341 ggcaacagta cttcaaacag aactgcagtc agcatgaatc cagtccagat atttcacatt
     2401 ttgaaaggtc aaagcaagtt tcatcagtca atgaggaaga ttttgttaga cttaaacagc
     2461 agatttcaga tcatatttca caatcctgtg gatctgggca aatgaaaatg tttcaggaag
     2521 tttctgcagc agatgctttt ggtccaggta ctgagggaca agtagaaaga tttgaaacag
     2581 ttggcatgga ggctgcgact gatgaaggca tgcctaaaag ttacttacca cagactgtac
     2641 ggcaaggcgg ctacatgcct cagtaggcgg ccgctcgagt ctagagggcc cgtttaaacc
     2701 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2761 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2821 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2881 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2941 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3001 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3061 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3121 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3181 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3241 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3301 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3361 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3421 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3481 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3541 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3601 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3661 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3721 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     3781 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3841 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3901 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3961 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4021 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4081 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4141 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4201 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4261 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4321 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4381 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4441 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4501 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4561 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4621 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4681 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4741 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4801 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4861 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4921 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     4981 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5041 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5101 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5161 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5221 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5281 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5341 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5401 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5461 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5521 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5581 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5641 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5701 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5761 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5821 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5881 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5941 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6001 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6061 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6121 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6181 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6241 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6301 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6361 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6421 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6481 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6541 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6601 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6661 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6721 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6781 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6841 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6901 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6961 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7021 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7081 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     7141 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     7201 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     7261 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     7321 cttcgcgatg tacgggccag atat
//
