LOCUS       40924_44339        7314 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTS1004, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7314)
  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 7314)
  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 7314)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7314)
  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..7314
                     /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    1808..2636
                     /label=IL6ST
                     /note="IL6ST"
     polyA_signal    2687..2911
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2957..3385
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3399..3728
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3795..4586
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4763..4896
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4933..4949)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4957..4973)
                     /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(4981..5011)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5026..5047)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5335..5920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6094..6951)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6952..7056)
                     /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 tgtccgcagc
     1801 agcagcaaat aagcgagacc taattaaaaa acacatctgg cctaatgttc cagatccttc
     1861 aaagagtcat attgcccagt ggtcacctca cactcctcca aggcacaatt ttaattcaaa
     1921 agatcaaatg tattcagatg gcaatttcac tgatgtaagt gttgtggaaa tagaagcaaa
     1981 tgacaaaaag ccttttccag aagatctgaa atcattggac ctgttcaaaa aggaaaaaat
     2041 taatactgaa ggacacagca gtggtattgg ggggtcttca tgcatgtcat cttctaggcc
     2101 aagcatttct agcagtgatg aaaatgaatc ttcacaaaac acttcgagca ctgtccagta
     2161 ttctaccgtg gtacacagtg gctacagaca ccaagttccg tcagtccaag tcttctcaag
     2221 atccgagtct acccagccct tgttagattc agaggagcgg ccagaagatc tacaattagt
     2281 agatcatgta gatggcggtg atggtatttt gcccaggcaa cagtacttca aacagaactg
     2341 cagtcagcat gaatccagtc cagatatttc acattttgaa aggtcaaagc aagtttcatc
     2401 agtcaatgag gaagattttg ttagacttaa acagcagatt tcagatcata tttcacaatc
     2461 ctgtggatct gggcaaatga aaatgtttca ggaagtttct gcagcagatg cttttggtcc
     2521 aggtactgag ggacaagtag aaagatttga aacagttggc atggaggctg cgactgatga
     2581 aggcatgcct aaaagttact taccacagac tgtacggcaa ggcggctaca tgcctcgcgg
     2641 ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     2701 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2761 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2821 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2881 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     2941 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3001 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3061 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3121 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3181 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3241 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3301 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     3361 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3421 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3481 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3541 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3601 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3661 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3721 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     3781 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     3841 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     3901 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     3961 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4021 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4081 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4141 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4201 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4261 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4321 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4381 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4441 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4501 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4561 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     4621 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     4681 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     4741 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     4801 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4861 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     4921 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4981 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5041 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5101 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5161 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5221 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5281 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5341 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5401 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5461 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5521 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5581 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5641 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5701 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5761 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5821 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     5881 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5941 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6001 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6061 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6121 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6181 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6241 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6301 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6361 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6421 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6481 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6541 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6601 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6661 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6721 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6781 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6841 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6901 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6961 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7021 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7081 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     7141 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     7201 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     7261 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atat
//
