LOCUS       40924_28052        7338 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLeo623, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7338)
  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 7338)
  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 7338)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7338)
  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..7338
                     /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..314
                     /label=heavy
                     /note="heavy"
     misc_feature    315..650
                     /label=light
                     /note="light"
     misc_feature    711..1061
                     /label=heavy
                     /note="heavy"
     misc_feature    1074..1817
                     /label=EpoR-D1_D2_TM
                     /note="EpoR-D1_D2_TM"
     misc_feature    1347..1349
                     /label=F93A
                     /note="F93A"
     misc_feature    1749..1817
                     /label=transmembrane
                     /note="transmembrane"
     misc_feature    1827..2655
                     /label=IL6ST
                     /note="IL6ST"
     misc_feature    2178..2180
                     /label=Y759A
                     /note="Y759A"
     polyA_signal    2711..2935
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2981..3409
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3423..3752
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3819..4610
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4787..4920
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4957..4973)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4981..4997)
                     /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(5005..5035)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5050..5071)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5359..5944)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6118..6975)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6976..7080)
                     /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 atccgacatc gttcttacgc aatctcctcc gtccttggct gtgagcctgg
      361 gtcaacgtgc caccatctca tgtcgtgcga gcgagagtat tgatttgtac ggattcacat
      421 ttatgcactg gtatcagcaa aagcccggtc aaccacccaa aattctgatt taccgcgcgt
      481 caaatttaga gtcaggcatt cctgcgcgct tctcaggttc gggctcacgc acagacttta
      541 cattgacaat taacccagtg gaagcagacg atgttgcgac atattattgt caacagacgc
      601 acgaagatcc atacacattt ggtggcggca ctaaattgga aattaagcgt ggtggtggag
      661 gatcaggtgg tggagggtct ggcggaggcg gtagcggtgg aggtggaagc caagtacagt
      721 tacagcagag tggcgcagag ttggcaaaac cgggagcctc ggttaagatg tcatgcaaga
      781 cgagtggata ctcatttagc agttactgga tgcactgggt taaacaacgc ccagggcaag
      841 gacttgaatg gattgggtat atcaatccat cgaccggata tacagagaac aaccagaagt
      901 ttaaagacaa ggttactctg accgctgaca agtcaagtaa cacggcatat atgcaattaa
      961 acagccttac ctctgaagat tcagccgtct attactgtgc tcgctcaggg cgtctgtact
     1021 tcgatgtatg gggggcaggt acgacagtga cggtgtctag ttccggagaa ttcgcacctt
     1081 cacccagcct cccggacccc aagtttgaga gcaaagcggc cctgctggca tcccggggct
     1141 ccgaagaact tctgtgcttc acccaacgct tggaagactt ggtgtgtttc tgggaggaag
     1201 cggcgagctc cgggatggac ttcaactaca gcttctcata ccagctcgag ggtgagtcac
     1261 gaaagtcatg tagcctgcac caggctccca ccgtccgcgg ctccgtgcgt ttctggtgtt
     1321 cactgccaac agcggacaca tcgagtgctg tgccgctgga gctgcaggtg acggaggcgt
     1381 ccggttctcc tcgctatcac cgcatcatcc atatcaatga agtagtgctc ctggacgccc
     1441 ccgcggggct gctggcgcgc cgggcagaag agggcagcca cgtggtgctg cgctggctgc
     1501 cacctcctgg agcacctatg accacccaca tccgatatga agtggacgtg tcggcaggca
     1561 accgggcagg agggacacaa agggtggagg tcctggaagg ccgcactgag tgtgttctga
     1621 gcaacctgcg gggcgggacg cgctacacct tcgctgttcg agcgcgcatg gccgagccga
     1681 gcttcagcgg attctggagt gcctggtctg agcccgcgtc actactgacc gctagcgacc
     1741 tggaccctct catcttgacg ctgtctctca ttctggtcct catctcgttg ttgctgacgg
     1801 ttctggccct gctgtccgca gcagcaaata agcgagacct aattaaaaaa cacatctggc
     1861 ctaatgttcc agatccttca aagagtcata ttgcccagtg gtcacctcac actcctccaa
     1921 ggcacaattt taattcaaaa gatcaaatgt attcagatgg caatttcact gatgtaagtg
     1981 ttgtggaaat agaagcaaat gacaaaaagc cttttccaga agatctgaaa tcattggacc
     2041 tgttcaaaaa ggaaaaaatt aatactgaag gacacagcag tggtattggg gggtcttcat
     2101 gcatgtcatc ttctaggcca agcatttcta gcagtgatga aaatgaatct tcacaaaaca
     2161 cttcgagcac tgtccaggct tctaccgtgg tacacagtgg ctacagacac caagttccgt
     2221 cagtccaagt cttctcaaga tccgagtcta cccagccctt gttagattca gaggagcggc
     2281 cagaagatct acaattagta gatcatgtag atggcggtga tggtattttg cccaggcaac
     2341 agtacttcaa acagaactgc agtcagcatg aatccagtcc agatatttca cattttgaaa
     2401 ggtcaaagca agtttcatca gtcaatgagg aagattttgt tagacttaaa cagcagattt
     2461 cagatcatat ttcacaatcc tgtggatctg ggcaaatgaa aatgtttcag gaagtttctg
     2521 cagcagatgc ttttggtcca ggtactgagg gacaagtaga aagatttgaa acagttggca
     2581 tggaggctgc gactgatgaa ggcatgccta aaagttactt accacagact gtacggcaag
     2641 gcggctacat gcctcagtag gcggccgctc gagtctagag ggcccgttta aacccgctga
     2701 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2761 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2821 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2881 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2941 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3001 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3061 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3121 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3181 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3241 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3301 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3361 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3421 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3481 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3541 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3601 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3661 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3721 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     3781 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3841 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3901 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3961 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4021 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4081 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4141 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4201 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4261 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4321 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     4381 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4441 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4501 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4561 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4621 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4681 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4741 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4801 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4861 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4921 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4981 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5041 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5101 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5161 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5221 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5281 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5341 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5401 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5461 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5521 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5581 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5641 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5701 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5761 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5821 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5881 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5941 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6001 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6061 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6121 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6181 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6241 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6301 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6361 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6421 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6481 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6541 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6601 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6661 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6721 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6781 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6841 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6901 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6961 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7021 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7081 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     7141 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     7201 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     7261 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     7321 gatgtacggg ccagatat
//
