LOCUS       40924_44329        7308 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTS1002, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7308)
  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 7308)
  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 7308)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7308)
  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..7308
                     /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    1802..2630
                     /label=IL6ST
                     /note="IL6ST"
     polyA_signal    2681..2905
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2951..3379
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3393..3722
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3789..4580
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4757..4890
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4927..4943)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4951..4967)
                     /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(4975..5005)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5020..5041)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5329..5914)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6088..6945)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6946..7050)
                     /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 aaataagcga gacctaatta aaaaacacat ctggcctaat gttccagatc cttcaaagag
     1861 tcatattgcc cagtggtcac ctcacactcc tccaaggcac aattttaatt caaaagatca
     1921 aatgtattca gatggcaatt tcactgatgt aagtgttgtg gaaatagaag caaatgacaa
     1981 aaagcctttt ccagaagatc tgaaatcatt ggacctgttc aaaaaggaaa aaattaatac
     2041 tgaaggacac agcagtggta ttggggggtc ttcatgcatg tcatcttcta ggccaagcat
     2101 ttctagcagt gatgaaaatg aatcttcaca aaacacttcg agcactgtcc agtattctac
     2161 cgtggtacac agtggctaca gacaccaagt tccgtcagtc caagtcttct caagatccga
     2221 gtctacccag cccttgttag attcagagga gcggccagaa gatctacaat tagtagatca
     2281 tgtagatggc ggtgatggta ttttgcccag gcaacagtac ttcaaacaga actgcagtca
     2341 gcatgaatcc agtccagata tttcacattt tgaaaggtca aagcaagttt catcagtcaa
     2401 tgaggaagat tttgttagac ttaaacagca gatttcagat catatttcac aatcctgtgg
     2461 atctgggcaa atgaaaatgt ttcaggaagt ttctgcagca gatgcttttg gtccaggtac
     2521 tgagggacaa gtagaaagat ttgaaacagt tggcatggag gctgcgactg atgaaggcat
     2581 gcctaaaagt tacttaccac agactgtacg gcaaggcggc tacatgcctc gcggccgctc
     2641 gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     2701 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2761 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     2821 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     2881 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     2941 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3001 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3061 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3121 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3181 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3241 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3301 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3361 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3421 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3481 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3541 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3601 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3661 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3721 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     3781 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3841 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3901 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3961 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4021 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4081 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4141 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4201 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4261 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     4321 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4381 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4441 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4501 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4561 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     4621 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4681 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4741 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4801 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4861 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     4921 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4981 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5041 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5101 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5161 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5221 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5281 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5341 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5401 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5461 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5521 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5581 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5641 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5701 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5761 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5821 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa
     5881 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5941 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6001 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6061 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6121 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6181 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6241 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6301 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6361 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6421 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6481 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6541 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6601 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6661 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6721 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6781 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6841 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6901 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6961 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7021 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7081 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
     7141 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     7201 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     7261 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatat
//
