LOCUS       62056_17430        5926 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pMSx88, complete sequence.
ACCESSION   MT267327
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5926)
  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 5926)
  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 5926)
  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..5926
                     /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"
     misc_feature    327..332
                     /label=Kozak+ATG
                     /note="Kozak+ATG"
     CDS             330..1478
                     /codon_start=1
                     /transl_table=11
                     /product="truncated NarX"
                     /label=truncated NarX
                     /protein_id="QKE44366.1"
                     /translation="MTSVLEQRVQEKTAGLEHKNQILSFLWQANRRLHSRAPLCERLSP
                     VLNGLQNLTLLRDIELRVYDTDDEENHQEFTCQPDMTCDDKGCQLCPRGVLPVGDRGTT
                     LKWRLADSHTQYGILLATLPQGRHLSHDQQQLVDTLVEQLTATLALDRHQERQQQLIVM
                     EERATIARELHDSIAQSLSCMKMQVSCLQMQGDALPESSRELLSQIRNELNASWAQLRE
                     LLTTFRLQLTEPGLRPALEASCEEYSAKFGFPVKLDYQLPPRLVPSHQAIHLLQIAREA
                     LSNALKHSQASEVVVTVAQNDNQVKLTVQDNGCGVPENAIRSNHYGMIIMRDRAQSLRG
                     DCRVRRRESGGTEVVVTFIPEKTFTDVQGDTHEASASGSTGV"
     misc_feature    441..1448
                     /label=NarX 263-598
                     /note="NarX 263-598"
     misc_feature    826..1448
                     /label=Kinase Domain
                     /note="Kinase Domain"
     misc_feature    849..851
                     /label=pH
                     /note="pH"
     polyA_signal    1515..1739
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1785..2213
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2227..2556
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2623..3414
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    3591..3724
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3761..3777)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3785..3801)
                     /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(3809..3839)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3854..3875)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4163..4748)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4922..5779)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5780..5884)
                     /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 tgactagtgt acttgagcag cgggttcagg
      361 agaaaaccgc cgggctggag cataaaaatc agatcctctc ttttttatgg caggctaacc
      421 gccgtttgca ttcccgcgcc ccgctgtgtg aacgcctgtc acctgtactc aacggcttac
      481 agaatttaac cctgctacgt gatatcgaat tgcgggtgta tgacactgat gatgaagaga
      541 atcatcagga gtttacctgc cagccagata tgacttgtga tgataaaggc tgccagctct
      601 gcccgcgcgg cgtattaccc gttggcgatc gcggcacaac cctgaagtgg cggctggctg
      661 actcacatac gcagtacggt attttgctgg cgaccctacc gcaggggcgt catcttagcc
      721 atgatcaaca acaactggtg gataccctgg ttgaacaact caccgccacg ctggcgctgg
      781 atcggcatca ggaacgtcag caacagttga tcgtgatgga agagcgtgcc accattgcgc
      841 gcgaactgca tgattctatt gcccaatctc tctcttgcat gaagatgcag gtgagttgtt
      901 tacagatgca gggcgatgcg ctgccagaaa gcagccgcga actgttaagt cagatccgta
      961 acgaactgaa tgcatcctgg gcgcagttgc gtgaattgct caccacattc cgcttgcagc
     1021 tcaccgagcc tggattacgt ccggcgctgg aagcgagttg cgaagagtac agcgccaaat
     1081 ttggcttccc ggtgaagctg gattatcaat tgccgcctcg tctggtgcct tcacatcagg
     1141 caatccactt gttgcaaatt gcccgtgagg cattaagtaa cgccctcaaa cattcgcaag
     1201 cgagtgaggt cgtggtgacg gtggcgcaaa acgataatca ggtcaaactg accgtccagg
     1261 ataacggctg cggcgtgcct gaaaatgcca tccgcagcaa tcactacggc atgataataa
     1321 tgcgcgaccg tgcgcaaagt ttacgaggcg attgccgcgt ccgccgtcgt gaatcaggtg
     1381 gcaccgaagt ggtggtcacc tttattcccg aaaaaacttt cacagacgtc caaggagata
     1441 cccatgaggc tagtgctagc ggatccaccg gtgtctagaa agctttgagg ccggcctgca
     1501 gggatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     1561 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     1621 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     1681 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     1741 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     1801 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     1861 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     1921 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     1981 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2041 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2101 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     2161 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     2221 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2281 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     2341 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     2401 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     2461 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     2521 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     2581 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     2641 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     2701 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     2761 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     2821 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     2881 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     2941 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     3001 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     3061 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     3121 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     3181 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     3241 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     3301 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     3361 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     3421 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     3481 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     3541 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     3601 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3661 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     3721 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3781 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3841 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3901 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3961 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4021 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4081 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4141 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4201 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4261 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4321 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4381 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4441 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4501 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4561 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4621 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4681 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4741 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4801 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4861 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4921 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4981 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5041 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5101 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5161 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5221 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5281 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5341 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5401 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5461 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5521 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5581 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5641 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5701 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5761 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5821 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5881 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcggg
//
