LOCUS       62056_17380        5833 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pMSx132, complete sequence.
ACCESSION   MT267329
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5833)
  AUTHORS   Scheller L, Schmollack M, Bertschi A, Mansouri M, Saxena P, 
            Fussenegger M.
  TITLE     Phosphoregulated orthogonal signal transduction in mammalian cells
  JOURNAL   Nat Commun 11 (1), 3085 (2020)
  PUBMED    32555187
REFERENCE   2  (bases 1 to 5833)
  AUTHORS   Scheller L.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-MAR-2020) STI IBI-STI LPDI, EPFL, AI 3123 (Batiment 
            AI), Station 19, Lausanne 1015, Switzerland
REFERENCE   3  (bases 1 to 5833)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun"; date: "2020"; volume: "11"; issue: "1"; pages: "3085"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-MAR-2020) STI IBI-STI LPDI, EPFL, AI 3123 (Batiment AI), Station
            19, Lausanne 1015, Switzerland"
FEATURES             Location/Qualifiers
     source          1..5833
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    27..230
                     /label=SV40 early promoter
                     /note="SV40 early promoter"
     promoter        280..298
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             330..1385
                     /codon_start=1
                     /transl_table=11
                     /product="EnvZ-ORK"
                     /label=EnvZ-ORK
                     /protein_id="QKE44368.1"
                     /translation="MTSGSQVQLVESGGGLVQAGGSLRLSCTASGRTGTIYSMAWFRQA
                     PGKEREFLATVGWSSGITYYMDSVKGRFTISRDKGKNTVYLQMDSLKPEDTAVYYCTAT
                     RAYSVGYDYWGQGTQVTVSSASDDRTLLMAGVSHDLRTPLTRIRLATEMMSEQDGYLAE
                     SINKDIEECNAIIEQFIDYLRTASMEMADLNAVLGEVIAAESGYEREIETALYPGSIEV
                     KMHPLSIKRAVANMVVNAARYGNGWIKVSSGTEPNRAWFQVEDDGPGIAPEQRKHLFQP
                     FVRGDSARTISGTGLGLAIVQRIVDNHNGMLELGTSERGGLSIRAWLPVPVTRAQGTTK
                     EGASASGSTGV"
     misc_feature    339..701
                     /label=aCaffVHH
                     /note="aCaffVHH"
     misc_feature    420..443
                     /label=CDR1
                     /note="CDR1"
     misc_feature    492..515
                     /label=CDR2
                     /note="CDR2"
     misc_feature    633..668
                     /label=CDR3
                     /note="CDR3"
     misc_feature    708..1355
                     /note="EnvZ 232-450;GQEMP:AS"
     misc_feature    708..885
                     /label=envZ DHp
                     /note="envZ DHp"
     misc_feature    891..1355
                     /label=envZ CA
                     /note="envZ CA"
     polyA_signal    1422..1646
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1692..2120
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2134..2463
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2530..3321
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    3498..3631
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3668..3684)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3692..3708)
                     /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(3716..3746)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3761..3782)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4070..4655)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4829..5686)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5687..5791)
                     /label=AmpR promoter
ORIGIN
        1 agatctccac gcgtggtacc ctcgaggcga tctgcatctc aattagtcag caaccatagt
       61 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
      121 ccatcgctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
      181 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa gagctctctg
      241 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
      301 gacccaagct gttcgaagcg gaattcacca tgactagtgg ctctcaggtt caattggtgg
      361 aatctggagg gggtctcgta caggcaggcg gttctctccg actgagttgc acagcctccg
      421 gtaggactgg gaccatctac tcaatggcct ggtttcgcca ggccccaggc aaagaaagag
      481 agtttcttgc cactgtaggt tggagttctg ggatcacata ctacatggat tcagttaaag
      541 gaagattcac tatcagccga gataaaggga aaaatactgt gtacctccag atggactctc
      601 tgaaaccgga ggacacggct gtctactact gtacagccac ccgcgcctac tccgtagggt
      661 atgactactg ggggcaagga acacaggtaa ccgtctctag cgctagtgat gaccgcacgc
      721 tgctgatggc gggggtaagt cacgacttgc gcacgccgct gacgcgtatt cgcctggcga
      781 ctgagatgat gagcgagcag gatggctatc tggcagaatc gatcaataaa gatatcgaag
      841 agtgcaacgc catcattgag cagtttatcg actacctgcg caccgctagt atggaaatgg
      901 cggatcttaa tgcagtactc ggtgaggtga ttgctgccga aagtggctat gagcgggaaa
      961 ttgaaaccgc gctttacccc ggcagcattg aagtgaaaat gcacccgctg tcgatcaaac
     1021 gcgcggtggc gaatatggtg gtcaacgccg cccgttatgg caatggctgg atcaaagtca
     1081 gcagcggaac ggagccgaat cgcgcctggt tccaggtgga agatgacggt ccgggaattg
     1141 cgccggaaca acgtaagcac ctgttccagc cgtttgtccg cggcgacagt gcgcgcacca
     1201 ttagcggcac gggattaggg ctggcaattg tgcagcgtat cgtggataac cataacggga
     1261 tgctggagct tggcaccagc gagcggggcg ggctttccat tcgcgcctgg ctgccagtgc
     1321 cggtaacgcg ggcgcagggc acgacaaaag aaggggctag tgctagcgga tccaccggtg
     1381 tctagaaagc tttgaggccg gcctgcaggg atcagcctcg actgtgcctt ctagttgcca
     1441 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     1501 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     1561 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     1621 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     1681 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     1741 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     1801 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     1861 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     1921 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     1981 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     2041 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     2101 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     2161 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     2221 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2281 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     2341 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     2401 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     2461 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     2521 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     2581 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     2641 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     2701 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     2761 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     2821 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     2881 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     2941 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3001 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     3061 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3121 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3181 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3241 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     3301 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     3361 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     3421 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     3481 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3541 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3601 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     3661 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     3721 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     3781 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3841 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3901 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3961 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4021 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4081 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4141 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4201 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4261 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4321 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4381 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4441 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4501 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4561 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     4621 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4681 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4741 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4801 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4861 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4921 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4981 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5041 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5101 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5161 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5221 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5281 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5341 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5401 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5461 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5521 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5581 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5641 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5701 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5761 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5821 tcgacggatc ggg
//
