LOCUS       62056_17400        6250 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pMSx4, complete sequence.
ACCESSION   MT267321
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6250)
  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 6250)
  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 6250)
  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..6250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(4..861)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(862..966)
                     /label=AmpR promoter
     misc_feature    1029..1049
                     /label=OmpR binding element 2
                     /note="OmpR binding element 2"
     misc_feature    1060..1080
                     /label=OmpR binding element 1
                     /note="OmpR binding element 1"
     promoter        1094..1132
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        1177..1195
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1224..1229
                     /label=Kozak+ATG
                     /note="Kozak+ATG"
     CDS             1236..2732
                     /label=SEAP
                     /note="secreted alkaline phosphatase from human placenta"
     polyA_signal    2847..3071
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3117..3545
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3559..3888
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3955..4746
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    4923..5056
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5093..5109)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5117..5133)
                     /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(5141..5171)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5186..5207)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5495..6080)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
       61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
      121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
      181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
      661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
      721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
      781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
      841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
      901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
      961 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctccacgc
     1021 gtctcgagat ttacattttg aaacatctat agcgccggca tttacatttt gaaacatcta
     1081 gctgcaggtc gagggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct
     1141 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
     1201 ccaagctgtt cgaagcggaa ttcaccatga ctagtctgct gctgctgctg ctgggcctga
     1261 gactgcagct gagcctgggc atcatccccg tggaggagga gaaccccgac ttctggaaca
     1321 gagaggccgc cgaggccctg ggcgccgcca agaagctgca gcccgcccag accgccgcca
     1381 agaacctgat catcttcctg ggcgacggca tgggcgtgag caccgtgacc gccgccagaa
     1441 tcctgaaggg ccagaagaag gacaagctgg gccccgagat ccccctggcc atggacagat
     1501 tcccctacgt ggccctgagc aagacctaca acgtggacaa gcacgtgccc gacagcggcg
     1561 ccaccgccac cgcctacctg tgcggcgtga agggcaactt ccagaccatc ggcctgagcg
     1621 ccgccgccag attcaaccag tgcaacacca ccagaggcaa cgaggtgatc agcgtgatga
     1681 acagagccaa gaaggccggc aagagcgtgg gcgtggtgac caccaccaga gtgcagcacg
     1741 ccagccccgc cggcacctac gcccacaccg tgaacagaaa ctggtacagc gacgccgacg
     1801 tgcccgccag cgccagacag gagggctgcc aggacatcgc cacccagctg atcagcaaca
     1861 tggacatcga cgtgatcctg ggcggcggca gaaagtacat gttcagaatg ggcacccccg
     1921 accccgagta ccccgacgac tacagccagg gcggcaccag actggacggc aagaacctgg
     1981 tgcaggagtg gctggccaag agacagggcg ccagatacgt gtggaacaga accgagctga
     2041 tgcaggccag cctggacccc agcgtgaccc acctgatggg cctgttcgag cccggcgaca
     2101 tgaagtacga gatccacaga gacagcaccc tggaccccag cctgatggag atgaccgagg
     2161 ccgccctgag actgctgagc agaaacccca gaggcttctt cctgttcgtg gagggcggca
     2221 gaatcgacca cggccaccac gagagcagag cctacagagc cctgaccgag accatcatgt
     2281 tcgacgacgc catcgagaga gccggccagc tgaccagcga ggaggacacc ctgagcctgg
     2341 tgaccgccga ccacagccac gtgttcagct tcggcggcta ccccctgaga ggcagcagca
     2401 tcttcggcct ggcccccggc aaggccagag acagaaaggc ctacaccgtg ctgctgtacg
     2461 gcaacggccc cggctacgtg ctgaaggacg gcgccagacc cgacgtgacc gagagcgaga
     2521 gcggcagccc cgagtacaga cagcagagcg ccgtgcccct ggacgaggag acccacgccg
     2581 gcgaggacgt ggccgtgttc gccagaggcc cccaggccca cctggtgcac ggcgtgcagg
     2641 agcagacctt catcgcccac gtgatggcct tcgccgcctg cctggagccc tacaccgcct
     2701 gcgacctggc cccccccgcc ggcaccaccg acgccgccca ccccggctac agcagagtgg
     2761 gcgccgccgg cagattcgag cagaccgcta gcggatccac cggtgtctag aaagctttga
     2821 ggccggcctg cagggatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     2881 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2941 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3001 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3061 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3121 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3181 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3241 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3301 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3361 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3421 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3481 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3541 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3601 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3661 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3721 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3781 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3841 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3901 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3961 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4021 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4081 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4141 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4201 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4261 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4321 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4381 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4441 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4501 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4561 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4621 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4681 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4741 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4801 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4861 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4921 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4981 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5041 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5101 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5161 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5221 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5281 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5341 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5401 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5461 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5521 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5581 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5641 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5701 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5761 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5821 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5881 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5941 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6001 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     6061 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6121 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6181 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6241 ggtctgacag
//
