LOCUS       40924_44334        7311 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTS1003, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7311)
  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 7311)
  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 7311)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7311)
  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..7311
                     /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    1805..2633
                     /label=IL6ST
                     /note="IL6ST"
     polyA_signal    2684..2908
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2954..3382
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3396..3725
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3792..4583
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4760..4893
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4930..4946)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4954..4970)
                     /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(4978..5008)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5023..5044)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5332..5917)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6091..6948)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6949..7053)
                     /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 agcaaataag cgagacctaa ttaaaaaaca catctggcct aatgttccag atccttcaaa
     1861 gagtcatatt gcccagtggt cacctcacac tcctccaagg cacaatttta attcaaaaga
     1921 tcaaatgtat tcagatggca atttcactga tgtaagtgtt gtggaaatag aagcaaatga
     1981 caaaaagcct tttccagaag atctgaaatc attggacctg ttcaaaaagg aaaaaattaa
     2041 tactgaagga cacagcagtg gtattggggg gtcttcatgc atgtcatctt ctaggccaag
     2101 catttctagc agtgatgaaa atgaatcttc acaaaacact tcgagcactg tccagtattc
     2161 taccgtggta cacagtggct acagacacca agttccgtca gtccaagtct tctcaagatc
     2221 cgagtctacc cagcccttgt tagattcaga ggagcggcca gaagatctac aattagtaga
     2281 tcatgtagat ggcggtgatg gtattttgcc caggcaacag tacttcaaac agaactgcag
     2341 tcagcatgaa tccagtccag atatttcaca ttttgaaagg tcaaagcaag tttcatcagt
     2401 caatgaggaa gattttgtta gacttaaaca gcagatttca gatcatattt cacaatcctg
     2461 tggatctggg caaatgaaaa tgtttcagga agtttctgca gcagatgctt ttggtccagg
     2521 tactgaggga caagtagaaa gatttgaaac agttggcatg gaggctgcga ctgatgaagg
     2581 catgcctaaa agttacttac cacagactgt acggcaaggc ggctacatgc ctcgcggccg
     2641 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     2701 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2761 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2821 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2881 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     2941 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3001 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3061 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3121 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3181 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3241 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3301 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3361 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3421 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3481 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3541 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3601 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3661 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3721 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     3781 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3841 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3901 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3961 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4021 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4081 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4141 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4201 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4261 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4321 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4381 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4441 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4501 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4561 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4621 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4681 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4741 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4801 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4861 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     4921 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4981 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5041 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5101 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5161 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5221 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5281 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5341 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5401 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5461 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5521 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5581 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5641 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5701 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5761 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5821 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg
     5881 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5941 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6001 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6061 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6121 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6181 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6241 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6301 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6361 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6421 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6481 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6541 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6601 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6661 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6721 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6781 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6841 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6901 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6961 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7021 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7081 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     7141 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     7201 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     7261 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata t
//
