LOCUS       40924_44324        7305 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTS1001, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7305)
  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 7305)
  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 7305)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7305)
  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..7305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        8..387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        388..591
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        636..654
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    674
                     /label=MCS
                     /note="MCS"
     sig_peptide     680..742
                     /label=Ig-kappa leader
                     /note="leader sequence from mouse immunoglobulin kappa 
                     light
                     chain"
     misc_feature    743..757
                     /label=secretion
                     /note="secretion"
     CDS             767..1045
                     /codon_start=1
                     /label=FRB
                     /note="FKBP-rapamycin binding domain of human FRAP"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     misc_feature    1047..1795
                     /label=EpoR-D1_D2_TM
                     /note="EpoR-D1_D2_TM"
     misc_feature    1727..1795
                     /label=transmembrane
                     /note="transmembrane"
     misc_feature    1799..2627
                     /label=IL6ST
                     /note="IL6ST"
     polyA_signal    2678..2902
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2948..3376
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3390..3719
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3786..4577
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4754..4887
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4924..4940)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4948..4964)
                     /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(4972..5002)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5017..5038)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5326..5911)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6085..6942)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6943..7047)
                     /label=AmpR promoter
ORIGIN
        1 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      361 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      421 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      481 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      541 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
      601 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
      661 caagctgaac caagctagca tggagacaga cacactcctg ctatgggtac tgctgctctg
      721 ggttccaggt tccactggtg acggagatat acatatggcc tctcgcatcc tgtggcatga
      781 aatgtggcac gaaggtctgg aagaagcatc tcgcctgtat tttggcgagc gtaacgtgaa
      841 aggcatgttt gaagtgctgg agcctctgca cgcaatgatg gagcgtggac ctcaaactct
      901 gaaagagaca tccttcaatc aggcttatgg ccgtgatctg atggaggcac aggaatggtg
      961 tcgtaaatat atgaaaagcg ggaatgtgaa agacctgctg caagcatggg atctgtatta
     1021 tcatgtgttc cgccgtatca gcaaagaatt cgcaccttca cccagcctcc cggaccccaa
     1081 gtttgagagc aaagcggccc tgctggcatc ccggggctcc gaagaacttc tgtgcttcac
     1141 ccaacgcttg gaagacttgg tgtgtttctg ggaggaagcg gcgagctccg ggatggactt
     1201 caactacagc ttctcatacc agctcgaggg tgagtcacga aagtcatgta gcctgcacca
     1261 ggctcccacc gtccgcggct ccgtgcgttt ctggtgttca ctgccaacag cggacacatc
     1321 gagttttgtg ccgctggagc tgcaggtgac ggaggcgtcc ggttctcctc gctatcaccg
     1381 catcatccat atcaatgaag tagtgctcct ggacgccccc gcggggctgc tggcgcgccg
     1441 ggcagaagag ggcagccacg tggtgctgcg ctggctgcca cctcctggag cacctatgac
     1501 cacccacatc cgatatgaag tggacgtgtc ggcaggcaac cgggcaggag ggacacaaag
     1561 ggtggaggtc ctggaaggcc gcactgagtg tgttctgagc aacctgcggg gcgggacgcg
     1621 ctacaccttc gctgttcgag cgcgcatggc cgagccgagc ttcagcggat tctggagtgc
     1681 ctggtctgag cccgcgtcac tactgaccgc tagcgacctg gaccctctca tcttgacgct
     1741 gtctctcatt ctggtcctca tctcgttgtt gctgacggtt ctggccctgc tgtccgcaaa
     1801 taagcgagac ctaattaaaa aacacatctg gcctaatgtt ccagatcctt caaagagtca
     1861 tattgcccag tggtcacctc acactcctcc aaggcacaat tttaattcaa aagatcaaat
     1921 gtattcagat ggcaatttca ctgatgtaag tgttgtggaa atagaagcaa atgacaaaaa
     1981 gccttttcca gaagatctga aatcattgga cctgttcaaa aaggaaaaaa ttaatactga
     2041 aggacacagc agtggtattg gggggtcttc atgcatgtca tcttctaggc caagcatttc
     2101 tagcagtgat gaaaatgaat cttcacaaaa cacttcgagc actgtccagt attctaccgt
     2161 ggtacacagt ggctacagac accaagttcc gtcagtccaa gtcttctcaa gatccgagtc
     2221 tacccagccc ttgttagatt cagaggagcg gccagaagat ctacaattag tagatcatgt
     2281 agatggcggt gatggtattt tgcccaggca acagtacttc aaacagaact gcagtcagca
     2341 tgaatccagt ccagatattt cacattttga aaggtcaaag caagtttcat cagtcaatga
     2401 ggaagatttt gttagactta aacagcagat ttcagatcat atttcacaat cctgtggatc
     2461 tgggcaaatg aaaatgtttc aggaagtttc tgcagcagat gcttttggtc caggtactga
     2521 gggacaagta gaaagatttg aaacagttgg catggaggct gcgactgatg aaggcatgcc
     2581 taaaagttac ttaccacaga ctgtacggca aggcggctac atgcctcgcg gccgctcgag
     2641 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     2701 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2761 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2821 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     2881 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     2941 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3001 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3061 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3121 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3181 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3241 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3301 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3361 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3421 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3481 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3541 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3601 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     3661 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3721 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     3781 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     3841 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     3901 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3961 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4021 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4081 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4141 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4201 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4261 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     4321 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     4381 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     4441 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     4501 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     4561 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     4621 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4681 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4741 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4801 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4861 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     4921 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4981 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5041 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5101 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5161 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5221 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5281 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5341 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5401 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5461 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5521 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5581 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5641 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5701 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5761 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5821 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca
     5881 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5941 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6001 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6061 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6121 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6181 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6241 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6301 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6361 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6421 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6481 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6541 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6601 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6661 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6721 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6781 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6841 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6901 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6961 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7021 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     7081 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     7141 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     7201 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     7261 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatat
//
