LOCUS       62056_17270        5866 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pMS247, complete sequence.
ACCESSION   MT267320
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5866)
  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 5866)
  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 5866)
  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..5866
                     /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..1418
                     /codon_start=1
                     /transl_table=11
                     /product="ORK"
                     /label=ORK
                     /protein_id="QKE44359.1"
                     /translation="MTSGSQVQLVESGGGLVQAGGSLRLSCTASGRTGTIYSMAWFRQA
                     PGKEREFLATVGWSSGITYYMDSVKGRFTISRDKGKNTVYLQMDSLKPEDTAVYYCTAT
                     RAYSVGYDYWGQGTQVTVSSASGGGGSASKTEVRKLMQRLDGLVNYADALRERSHEFMN
                     KLHVILGLLHLKSYKQLEDYILKTANNYQEEIGSLLGKIKSPVIAGFLISKINRATDLG
                     HTLILNSESQLPDSGSEDQVATLITTLGNLIENALEALGPEPGGEISVTLHYRHGWLHC
                     EVNDDGPGIAPDKIDHIFDKGVSTKGSERGVGLALVKQQVENLGGSIAVESEPGIFTQF
                     FVQIPWDGERSNRASASGSTGV"
     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    729..1388
                     /label=DucS 324-543
                     /note="DucS 324-543"
     polyA_signal    1455..1679
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1725..2153
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2167..2496
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2563..3354
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    3531..3664
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3701..3717)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3725..3741)
                     /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(3749..3779)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3794..3815)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4103..4688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4862..5719)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5720..5824)
                     /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 cgctagtggc ggcggcggca
      721 gtgctagtaa aactgaagta cgtaaactga tgcagcgact cgacggtctg gtcaactatg
      781 ctgacgcact tcgtgaacga tcccacgaat ttatgaataa attgcatgtg attctcggat
      841 tattgcatct gaagagttat aagcagttgg aagattacat tctcaaaaca gccaataact
      901 atcaggaaga gattggctct ctgctgggta agatcaaatc tccggttatc gctggttttt
      961 taatcagcaa gattaaccgc gcgaccgatt taggccatac gctgatttta aacagtgaaa
     1021 gccagctgcc agacagcggc agtgaggacc aggtcgcgac gctgattacc acgttgggaa
     1081 atctgataga aaacgcgctg gaggcattag ggccggaacc cggaggcgaa attagcgtaa
     1141 cattgcacta ccgtcacggc tggctgcact gtgaagttaa tgatgatgga ccggggatcg
     1201 cacccgataa aatcgatcac atttttgaca aaggtgtctc gacaaaagga agcgagcgag
     1261 gcgtcggttt agcacttgtc aaacaacagg tagaaaatct cggcggcagc atcgccgtgg
     1321 aatcggaacc cgggattttc acacaatttt ttgtccagat accctgggac ggggagaggt
     1381 cgaacagagc tagtgctagc ggatccaccg gtgtctagaa agctttgagg ccggcctgca
     1441 gggatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     1501 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     1561 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     1621 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     1681 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     1741 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     1801 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     1861 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     1921 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     1981 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2041 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     2101 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     2161 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2221 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     2281 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     2341 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     2401 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     2461 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     2521 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     2581 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     2641 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     2701 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     2761 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     2821 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     2881 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     2941 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     3001 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     3061 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     3121 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     3181 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     3241 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     3301 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     3361 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     3421 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     3481 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     3541 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3601 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     3661 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3721 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3781 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3841 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3901 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3961 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4021 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4081 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4141 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4201 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4261 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4321 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4381 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4441 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4501 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4561 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4621 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4681 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4741 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4801 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4861 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4921 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4981 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5041 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5101 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5161 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5221 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5281 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5341 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5401 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5461 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5521 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5581 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5641 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5701 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5761 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5821 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcggg
//
