LOCUS       62056_17425        5473 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pMSx80, complete sequence.
ACCESSION   MT267326
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5473)
  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 5473)
  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 5473)
  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..5473
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..9
                     /label=SV40 polyA
                     /note="SV40 polyA"
     primer_bind     complement(49..65)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(73..89)
                     /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(97..127)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(142..163)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(451..1036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1210..2067)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(2068..2172)
                     /label=AmpR promoter
     misc_feature    2241..2444
                     /label=SV40 early promoter
                     /note="SV40 early promoter"
     promoter        2494..2512
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2541..2546
                     /label=Kozak+ATG
                     /note="Kozak+ATG"
     CDS             2544..3239
                     /codon_start=1
                     /transl_table=11
                     /product="truncated EnvZ"
                     /label=truncated EnvZ
                     /protein_id="QKE44365.1"
                     /translation="MTSDDRTLLMAGVSHDLRTPLTRIRLATEMMSEQDGYLAESINKD
                     IEECNAIIEQFIDYLRTGQEMPMEMADLNAVLGEVIAAESGYEREIETALYPGSIEVKM
                     HPLSIKRAVANMVVNAARYGNGWIKVSSGTEPNRAWFQVEDDGPGIAPEQRKHLFQPFV
                     RGDSARTISGTGLGLAIVQRIVDNHNGMLELGTSERGGLSIRAWLPVPVTRAQGTTKEG
                     ASASGSTGV"
     misc_feature    2553..3209
                     /label=envZ 232-450
                     /note="envZ 232-450"
     misc_feature    2730..2747
                     /label=linker acceptor-ATP binding
                     /note="linker acceptor-ATP binding"
     polyA_signal    3276..3500
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3546..3974
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3988..4317
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4384..5175
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    5352..5473
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
       61 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
      121 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
      181 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
      241 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
      301 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
      361 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
      421 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
      481 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
      541 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
      601 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
      661 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
      721 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
      781 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
      841 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
      901 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
      961 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     1021 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     1081 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     1141 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     1201 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1261 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1321 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     1381 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1441 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1501 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     1561 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     1621 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     1681 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     1741 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     1801 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     1861 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     1921 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     1981 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     2041 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     2101 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     2161 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     2221 ccacgcgtgg taccctcgag gcgatctgca tctcaattag tcagcaacca tagtcccgcc
     2281 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatcg
     2341 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     2401 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaagagctc tctggctaac
     2461 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
     2521 agctgttcga agcggaattc accatgacta gtgatgaccg cacgctgctg atggcggggg
     2581 taagtcacga cttgcgcacg ccgctgacgc gtattcgcct ggcgactgag atgatgagcg
     2641 agcaggatgg ctatctggca gaatcgatca ataaagatat cgaagagtgc aacgccatca
     2701 ttgagcagtt tatcgactac ctgcgcaccg ggcaggagat gccgatggaa atggcggatc
     2761 ttaatgcagt actcggtgag gtgattgctg ccgaaagtgg ctatgagcgg gaaattgaaa
     2821 ccgcgcttta ccccggcagc attgaagtga aaatgcaccc gctgtcgatc aaacgcgcgg
     2881 tggcgaatat ggtggtcaac gccgcccgtt atggcaatgg ctggatcaaa gtcagcagcg
     2941 gaacggagcc gaatcgcgcc tggttccagg tggaagatga cggtccggga attgcgccgg
     3001 aacaacgtaa gcacctgttc cagccgtttg tccgcggcga cagtgcgcgc accattagcg
     3061 gcacgggatt agggctggca attgtgcagc gtatcgtgga taaccataac gggatgctgg
     3121 agcttggcac cagcgagcgg ggcgggcttt ccattcgcgc ctggctgcca gtgccggtaa
     3181 cgcgggcgca gggcacgaca aaagaagggg ctagtgctag cggatccacc ggtgtctaga
     3241 aagctttgag gccggcctgc agggatcagc ctcgactgtg ccttctagtt gccagccatc
     3301 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3361 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3421 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3481 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3541 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3601 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3661 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3721 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3781 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3841 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3901 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3961 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4021 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4081 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4141 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4201 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4261 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4321 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4381 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4441 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4501 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4561 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4621 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4681 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4741 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4801 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4861 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     4921 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4981 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5041 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5101 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5161 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5221 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5281 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5341 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5401 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5461 tcaatgtatc tta
//
