LOCUS       40924_44364        7329 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTS1009, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7329)
  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 7329)
  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 7329)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7329)
  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..7329
                     /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"
     sig_peptide     674..736
                     /label=Ig-kappa leader
                     /note="leader sequence from mouse immunoglobulin kappa 
                     light
                     chain"
     CDS             737..1060
                     /codon_start=1
                     /label=FKBP
                     /note="human FK506-binding protein FKBP12"
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     misc_feature    1062..1810
                     /label=EpoR-D1_D2_TM
                     /note="EpoR-D1_D2_TM"
     misc_feature    1742..1810
                     /label=transmembrane
                     /note="transmembrane"
     misc_feature    1823..2651
                     /label=IL6ST
                     /note="IL6ST"
     polyA_signal    2702..2926
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2972..3400
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3414..3743
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3810..4601
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4778..4911
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4948..4964)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4972..4988)
                     /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(4996..5026)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5041..5062)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5350..5935)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6109..6966)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6967..7071)
                     /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 caagctggct agcatggaga cagacacact cctgctatgg gtactgctgc tctgggttcc
      721 aggttccact ggtgacatgg gcgttcaggt tgaaaccatt agcccgggag atggccgtac
      781 atttccgaaa cgtggccaga catgtgttgt tcactatacc ggcatgctgg aggatggtaa
      841 aaaatttgat agcagccgtg accgtaataa accgttcaaa ttcatgctgg gcaaacagga
      901 agtcattcgc ggttgggaag aaggtgtcgc acagatgagt gtgggtcaac gtgccaaact
      961 gacgattagt cctgactatg cctatggagc aacaggtcat ccagggatta ttccgcctca
     1021 cgctactctg gtctttgatg tggaactgct gaaactggaa gaattcgcac cttcacccag
     1081 cctcccggac cccaagtttg agagcaaagc ggccctgctg gcatcccggg gctccgaaga
     1141 acttctgtgc ttcacccaac gcttggaaga cttggtgtgt ttctgggagg aagcggcgag
     1201 ctccgggatg gacttcaact acagcttctc ataccagctc gagggtgagt cacgaaagtc
     1261 atgtagcctg caccaggctc ccaccgtccg cggctccgtg cgtttctggt gttcactgcc
     1321 aacagcggac acatcgagtt ttgtgccgct ggagctgcag gtgacggagg cgtccggttc
     1381 tcctcgctat caccgcatca tccatatcaa tgaagtagtg ctcctggacg cccccgcggg
     1441 gctgctggcg cgccgggcag aagagggcag ccacgtggtg ctgcgctggc tgccacctcc
     1501 tggagcacct atgaccaccc acatccgata tgaagtggac gtgtcggcag gcaaccgggc
     1561 aggagggaca caaagggtgg aggtcctgga aggccgcact gagtgtgttc tgagcaacct
     1621 gcggggcggg acgcgctaca ccttcgctgt tcgagcgcgc atggccgagc cgagcttcag
     1681 cggattctgg agtgcctggt ctgagcccgc gtcactactg accgctagcg acctggaccc
     1741 tctcatcttg acgctgtctc tcattctggt cctcatctcg ttgttgctga cggttctggc
     1801 cctgctgtcc gcagcagcag caaataagcg agacctaatt aaaaaacaca tctggcctaa
     1861 tgttccagat ccttcaaaga gtcatattgc ccagtggtca cctcacactc ctccaaggca
     1921 caattttaat tcaaaagatc aaatgtattc agatggcaat ttcactgatg taagtgttgt
     1981 ggaaatagaa gcaaatgaca aaaagccttt tccagaagat ctgaaatcat tggacctgtt
     2041 caaaaaggaa aaaattaata ctgaaggaca cagcagtggt attggggggt cttcatgcat
     2101 gtcatcttct aggccaagca tttctagcag tgatgaaaat gaatcttcac aaaacacttc
     2161 gagcactgtc cagtattcta ccgtggtaca cagtggctac agacaccaag ttccgtcagt
     2221 ccaagtcttc tcaagatccg agtctaccca gcccttgtta gattcagagg agcggccaga
     2281 agatctacaa ttagtagatc atgtagatgg cggtgatggt attttgccca ggcaacagta
     2341 cttcaaacag aactgcagtc agcatgaatc cagtccagat atttcacatt ttgaaaggtc
     2401 aaagcaagtt tcatcagtca atgaggaaga ttttgttaga cttaaacagc agatttcaga
     2461 tcatatttca caatcctgtg gatctgggca aatgaaaatg tttcaggaag tttctgcagc
     2521 agatgctttt ggtccaggta ctgagggaca agtagaaaga tttgaaacag ttggcatgga
     2581 ggctgcgact gatgaaggca tgcctaaaag ttacttacca cagactgtac ggcaaggcgg
     2641 ctacatgcct cgcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     2701 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2761 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2821 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2881 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     2941 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3001 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3061 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3121 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3181 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3241 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3301 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3361 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     3421 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3481 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3541 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3601 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3661 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     3721 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     3781 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3841 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3901 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3961 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4021 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4081 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4141 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4201 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4261 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4321 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     4381 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4441 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4501 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4561 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4621 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     4681 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4741 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4801 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4861 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     4921 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4981 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5041 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5101 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5161 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5221 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5281 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5341 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5401 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5461 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5521 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5581 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5641 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5701 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5761 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5821 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5881 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5941 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6001 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6061 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6121 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6181 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6241 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6301 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6361 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6421 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6481 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6541 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6601 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6661 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6721 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6781 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6841 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6901 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6961 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7021 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7081 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
     7141 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     7201 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     7261 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
     7321 gccagatat
//
