LOCUS       62056_17395        6214 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pMSx245, complete sequence.
ACCESSION   MT267332
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6214)
  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 6214)
  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 6214)
  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..6214
                     /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..1766
                     /codon_start=1
                     /transl_table=11
                     /product="NarX-ORK"
                     /label=NarX-ORK
                     /protein_id="QKE44371.1"
                     /translation="MTSGSQVQLVESGGGLVQAGGSLRLSCTASGRTGTIYSMAWFRQA
                     PGKEREFLATVGWSSGITYYMDSVKGRFTISRDKGKNTVYLQMDSLKPEDTAVYYCTAT
                     RAYSVGYDYWGQGTQVTVSSASGGGGSASPLCERLSPVLNGLQNLTLLRDIELRVYDTD
                     DEENHQEFTCQPDMTCDDKGCQLCPRGVLPVGDRGTTLKWRLADSHTQYGILLATLPQG
                     RHLSHDQQQLVDTLVEQLTATLALDRHQERQQQLIVMEERATIARELHDSIAQSLSCMK
                     MQVSCLQMQGDALPESSRELLSQIRNELNASWAQLRELLTTFRLQLTEPGLRPALEASC
                     EEYSAKFGFPVKLDYQLPPRLVPSHQAIHLLQIAREALSNALKHSQASEVVVTVAQNDN
                     QVKLTVQDNGCGVPENAIRSNHYGMIIMRDRAQSLRGDCRVRRRESGGTEVVVTFIPEK
                     TFTDVQGDTHEASASGSTGV"
     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..1736
                     /label=narX 263-598
                     /note="narX 263-598"
     misc_feature    1114..1736
                     /label=Kinase Domain
                     /note="Kinase Domain"
     misc_feature    1137..1139
                     /label=pH
                     /note="pH"
     polyA_signal    1803..2027
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2073..2501
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2515..2844
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2911..3702
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    3879..4012
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4049..4065)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4073..4089)
                     /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(4097..4127)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4142..4163)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4451..5036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5210..6067)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6068..6172)
                     /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 gtgctagtcc gctgtgtgaa cgcctgtcac ctgtactcaa cggcttacag aatttaaccc
      781 tgctacgtga tatcgaattg cgggtgtatg acactgatga tgaagagaat catcaggagt
      841 ttacctgcca gccagatatg acttgtgatg ataaaggctg ccagctctgc ccgcgcggcg
      901 tattacccgt tggcgatcgc ggcacaaccc tgaagtggcg gctggctgac tcacatacgc
      961 agtacggtat tttgctggcg accctaccgc aggggcgtca tcttagccat gatcaacaac
     1021 aactggtgga taccctggtt gaacaactca ccgccacgct ggcgctggat cggcatcagg
     1081 aacgtcagca acagttgatc gtgatggaag agcgtgccac cattgcgcgc gaactgcatg
     1141 attctattgc ccaatctctc tcttgcatga agatgcaggt gagttgttta cagatgcagg
     1201 gcgatgcgct gccagaaagc agccgcgaac tgttaagtca gatccgtaac gaactgaatg
     1261 catcctgggc gcagttgcgt gaattgctca ccacattccg cttgcagctc accgagcctg
     1321 gattacgtcc ggcgctggaa gcgagttgcg aagagtacag cgccaaattt ggcttcccgg
     1381 tgaagctgga ttatcaattg ccgcctcgtc tggtgccttc acatcaggca atccacttgt
     1441 tgcaaattgc ccgtgaggca ttaagtaacg ccctcaaaca ttcgcaagcg agtgaggtcg
     1501 tggtgacggt ggcgcaaaac gataatcagg tcaaactgac cgtccaggat aacggctgcg
     1561 gcgtgcctga aaatgccatc cgcagcaatc actacggcat gataataatg cgcgaccgtg
     1621 cgcaaagttt acgaggcgat tgccgcgtcc gccgtcgtga atcaggtggc accgaagtgg
     1681 tggtcacctt tattcccgaa aaaactttca cagacgtcca aggagatacc catgaggcta
     1741 gtgctagcgg atccaccggt gtctagaaag ctttgaggcc ggcctgcagg gatcagcctc
     1801 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     1861 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     1921 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     1981 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2041 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2101 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2161 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2221 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2281 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2341 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2401 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2461 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     2521 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2581 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2641 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     2701 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     2761 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     2821 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     2881 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     2941 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3001 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3061 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3121 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3181 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3241 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3301 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3361 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3421 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3481 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3541 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3601 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     3661 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     3721 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     3781 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     3841 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     3901 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3961 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4021 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4081 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4141 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4201 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4261 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4321 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4381 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4441 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4501 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4561 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4621 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4681 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4741 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4801 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4861 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4921 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4981 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5041 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5101 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5161 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5221 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5281 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5341 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5401 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5461 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5521 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5581 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5641 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5701 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5761 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5821 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5881 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5941 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6001 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6061 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6121 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6181 cccgaaaagt gccacctgac gtcgacggat cggg
//
