LOCUS       62056_17420        5824 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pMSx78, complete sequence.
ACCESSION   MT267325
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5824)
  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 5824)
  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 5824)
  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..5824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    28..231
                     /label=SV40 early promoter
                     /note="SV40 early promoter"
     promoter        281..299
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    328..333
                     /label=Kozak+ATG
                     /note="Kozak+ATG"
     CDS             331..1377
                     /codon_start=1
                     /transl_table=11
                     /product="truncated NarX"
                     /label=truncated NarX
                     /protein_id="QKE44364.1"
                     /translation="MTSPLCERLSPVLNGLQNLTLLRDIELRVYDTDDEENHQEFTCQP
                     DMTCDDKGCQLCPRGVLPVGDRGTTLKWRLADSHTQYGILLATLPQGRHLSHDQQQLVD
                     TLVEQLTATLALDRHQERQQQLIVMEERATIARELHDSIAQSLSCMKMQVSCLQMQGDA
                     LPESSRELLSQIRNELNASWAQLRELLTTFRLQLTEPGLRPALEASCEEYSAKFGFPVK
                     LDYQLPPRLVPSHQAIHLLQIAREALSNALKHSQASEVVVTVAQNDNQVKLTVQDNGCG
                     VPENAIRSNHYGMIIMRDRAQSLRGDCRVRRRESGGTEVVVTFIPEKTFTDVQGDTHEA
                     SASGSTGV"
     misc_feature    340..1347
                     /label=narX 263-598
                     /note="narX 263-598"
     misc_feature    725..1347
                     /label=Kinase Domain
                     /note="Kinase Domain"
     misc_feature    748..750
                     /label=pH
                     /note="pH"
     polyA_signal    1414..1638
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1684..2112
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2126..2455
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2522..3313
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    3490..3623
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3660..3676)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3684..3700)
                     /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(3708..3738)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3753..3774)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4062..4647)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4821..5678)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5679..5783)
                     /label=AmpR promoter
ORIGIN
        1 gagatctcca cgcgtggtac cctcgaggcg atctgcatct caattagtca gcaaccatag
       61 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
      121 cccatcgctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
      181 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa agagctctct
      241 ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact cactataggg
      301 agacccaagc tgttcgaagc ggaattcacc atgactagtc cgctgtgtga acgcctgtca
      361 cctgtactca acggcttaca gaatttaacc ctgctacgtg atatcgaatt gcgggtgtat
      421 gacactgatg atgaagagaa tcatcaggag tttacctgcc agccagatat gacttgtgat
      481 gataaaggct gccagctctg cccgcgcggc gtattacccg ttggcgatcg cggcacaacc
      541 ctgaagtggc ggctggctga ctcacatacg cagtacggta ttttgctggc gaccctaccg
      601 caggggcgtc atcttagcca tgatcaacaa caactggtgg ataccctggt tgaacaactc
      661 accgccacgc tggcgctgga tcggcatcag gaacgtcagc aacagttgat cgtgatggaa
      721 gagcgtgcca ccattgcgcg cgaactgcat gattctattg cccaatctct ctcttgcatg
      781 aagatgcagg tgagttgttt acagatgcag ggcgatgcgc tgccagaaag cagccgcgaa
      841 ctgttaagtc agatccgtaa cgaactgaat gcatcctggg cgcagttgcg tgaattgctc
      901 accacattcc gcttgcagct caccgagcct ggattacgtc cggcgctgga agcgagttgc
      961 gaagagtaca gcgccaaatt tggcttcccg gtgaagctgg attatcaatt gccgcctcgt
     1021 ctggtgcctt cacatcaggc aatccacttg ttgcaaattg cccgtgaggc attaagtaac
     1081 gccctcaaac attcgcaagc gagtgaggtc gtggtgacgg tggcgcaaaa cgataatcag
     1141 gtcaaactga ccgtccagga taacggctgc ggcgtgcctg aaaatgccat ccgcagcaat
     1201 cactacggca tgataataat gcgcgaccgt gcgcaaagtt tacgaggcga ttgccgcgtc
     1261 cgccgtcgtg aatcaggtgg caccgaagtg gtggtcacct ttattcccga aaaaactttc
     1321 acagacgtcc aaggagatac ccatgaggct agtgctagcg gatccaccgg tgtctagaaa
     1381 gctttgaggc cggcctgcag ggatcagcct cgactgtgcc ttctagttgc cagccatctg
     1441 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     1501 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     1561 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     1621 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     1681 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     1741 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     1801 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     1861 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     1921 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     1981 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     2041 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     2101 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     2161 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     2221 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2281 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     2341 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     2401 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     2461 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     2521 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     2581 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     2641 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     2701 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     2761 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     2821 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     2881 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     2941 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3001 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     3061 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3121 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     3181 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     3241 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     3301 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     3361 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     3421 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     3481 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3541 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3601 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     3661 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3721 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3781 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3841 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3901 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3961 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4021 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4081 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4141 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4201 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4261 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4321 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4381 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4441 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4501 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4561 agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag
     4621 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4681 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4741 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4801 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4861 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4921 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4981 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5041 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5101 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5161 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5221 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5281 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5341 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5401 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5461 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5521 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5581 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5641 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5701 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5761 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     5821 tcgg
//
