LOCUS       62056_17385        5791 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pMSx228, complete sequence.
ACCESSION   MT267330
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5791)
  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 5791)
  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 5791)
  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..5791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    201..334
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(371..387)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(395..411)
                     /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(419..449)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(464..485)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(773..1358)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1532..2389)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2390..2494)
                     /label=AmpR promoter
     promoter        2816..2834
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2884..3162
                     /codon_start=1
                     /label=FRB
                     /note="FKBP-rapamycin binding domain of human FRAP"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRISK"
     polyA_signal    3916..4140
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4186..4614
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4628..4957
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             join(5024..5791,1..24)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
ORIGIN
        1 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
       61 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
      121 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
      181 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
      241 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
      301 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
      361 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
      421 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
      481 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
      541 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
      601 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
      661 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
      721 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
      781 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
      841 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
      901 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
      961 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1021 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1081 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1141 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1201 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1261 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt
     1321 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1381 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1441 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     1501 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     1561 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1621 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     1681 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1741 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     1801 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     1861 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     1921 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     1981 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2041 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     2101 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     2161 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     2221 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2281 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2341 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     2401 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2461 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     2521 acgtcgacgg atcgggagat ctccacgcgt ggtaccctcg aggcgatctg catctcaatt
     2581 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     2641 ccgcccattc tccgccccat cgctgactaa ttttttttat ttatgcagag gccgaggccg
     2701 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     2761 gcaaaagagc tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata
     2821 cgactcacta tagggagacc caagctgttc gaagcggaat tcaccatgac tagtgcttcc
     2881 cgcatcctgt ggcatgaaat gtggcacgaa ggtctggaag aagcatctcg cctgtatttt
     2941 ggcgagcgta acgtgaaagg catgtttgaa gtgctggagc ctctgcacgc aatgatggag
     3001 cgtggacctc aaactctgaa agagacatcc ttcaatcagg cttatggccg tgatctgatg
     3061 gaggcacagg aatggtgtcg taaatatatg aaaagcggga atgtgaaaga cctgctgcaa
     3121 gcatgggatc tgtattatca tgtgttccgc cgtatcagca aagctagtgg cggcggcggc
     3181 agtgctagta aaactgaagt acgtaaactg atgcagcgac tcgacggtct ggtcaactat
     3241 gctgacgcac ttcgtgaacg atcccacgaa tttatgaata aattgcatgt gattctcgga
     3301 ttattgcatc tgaagagtta taagcagttg gaagattaca ttctcaaaac agccaataac
     3361 tatcaggaag agattggctc tctgctgggt aagatcaaat ctccggttat cgctggtttt
     3421 ttaatcagca agattaaccg cgcgaccgat ttaggccata cgctgatttt aaacagtgaa
     3481 agccagctgc cagacagcgg cagtgaggac caggtcgcga cgctgattac cacgttggga
     3541 aatctgatag aaaacgcgct ggaggcatta gggccggaac ccggaggcga aattagcgta
     3601 acattgcact accgtcacgg ctggctgcac tgtgaagtta atgatgatgg accggggatc
     3661 gcacccgata aaatcgatca catttttgac aaaggtgtct cgacaaaagg aagcgagcga
     3721 ggcgtcggtt tagcacttgt caaacaacag gtagaaaatc tcggcggcag catcgccgtg
     3781 gaatcggaac ccgggatttt cacacaattt tttgtccaga taccctggga cggggagagg
     3841 tcgaacagag ctagtgctag cggatccacc ggtgtctaga aagctttgag gccggcctgc
     3901 agggatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     3961 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     4021 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     4081 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     4141 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     4201 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4261 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4321 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4381 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4441 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4501 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     4561 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     4621 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4681 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4741 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4801 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4861 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     4921 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     4981 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     5041 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     5101 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     5161 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     5221 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     5281 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5341 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5401 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5461 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5521 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     5581 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5641 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5701 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5761 cgccgctccc gattcgcagc gcatcgcctt c
//
