LOCUS       40924_28027        6969 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo618, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6969)
  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 6969)
  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 6969)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6969)
  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..6969
                     /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..692
                     /label=anti RR120
                     /note="anti RR120"
     misc_feature    705..1448
                     /label=EpoR-D1_D2_TM
                     /note="EpoR-D1_D2_TM"
     misc_feature    1380..1448
                     /label=transmembrane
                     /note="transmembrane"
     misc_feature    1458..2288
                     /label=IL6ST
                     /note="IL6ST"
     misc_feature    1809..1811
                     /label=Y759A
                     /note="Y759A"
     polyA_signal    2342..2566
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2612..3040
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3054..3383
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3450..4241
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4418..4551
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4588..4604)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4612..4628)
                     /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(4636..4666)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4681..4702)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4990..5575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5749..6606)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6607..6711)
                     /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 atcgagtttt 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 agcagcaaat aagcgagacc taattaaaaa acacatctgg cctaatgttc
     1501 cagatccttc aaagagtcat attgcccagt ggtcacctca cactcctcca aggcacaatt
     1561 ttaattcaaa agatcaaatg tattcagatg gcaatttcac tgatgtaagt gttgtggaaa
     1621 tagaagcaaa tgacaaaaag ccttttccag aagatctgaa atcattggac ctgttcaaaa
     1681 aggaaaaaat taatactgaa ggacacagca gtggtattgg ggggtcttca tgcatgtcat
     1741 cttctaggcc aagcatttct agcagtgatg aaaatgaatc ttcacaaaac acttcgagca
     1801 ctgtccaggc ttctaccgtg gtacacagtg gctacagaca ccaagttccg tcagtccaag
     1861 tcttctcaag atccgagtct acccagccct tgttagattc agaggagcgg ccagaagatc
     1921 tacaattagt agatcatgta gatggcggtg atggtatttt gcccaggcaa cagtacttca
     1981 aacagaactg cagtcagcat gaatccagtc cagatatttc acattttgaa aggtcaaagc
     2041 aagtttcatc agtcaatgag gaagattttg ttagacttaa acagcagatt tcagatcata
     2101 tttcacaatc ctgtggatct gggcaaatga aaatgtttca ggaagtttct gcagcagatg
     2161 cttttggtcc aggtactgag ggacaagtag aaagatttga aacagttggc atggaggctg
     2221 cgactgatga aggcatgcct aaaagttact taccacagac tgtacggcaa ggcggctaca
     2281 tgcctcagta ggcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     2341 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2401 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2461 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2521 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     2581 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     2641 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     2701 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2761 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2821 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     2881 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     2941 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3001 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     3061 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3121 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3181 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3241 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3301 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     3361 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     3421 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3481 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3541 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3601 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     3661 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3721 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3781 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3841 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     3901 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     3961 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4021 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4081 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4141 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4201 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4261 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4321 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4381 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4441 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4501 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     4561 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4621 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4681 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4741 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4801 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4861 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4921 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4981 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5041 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5101 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5161 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5221 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5281 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5341 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5401 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5461 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5521 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5581 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5641 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5701 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5761 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5821 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5881 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5941 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6001 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6061 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6121 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6181 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6241 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6301 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6361 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6421 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6481 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6541 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6601 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6661 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6721 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
     6781 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     6841 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     6901 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
     6961 gccagatat
//
