LOCUS       62056_13975        5476 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pLeo1231, complete sequence.
ACCESSION   MT267304
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5476)
  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 5476)
  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 5476)
  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..5476
                     /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    1017
                     /label=Promoter Region
                     /note="Promoter Region"
     misc_feature    1035..1238
                     /label=SV40 early promoter
                     /note="SV40 early promoter"
     promoter        1288..1306
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1338..2036
                     /codon_start=1
                     /transl_table=11
                     /product="truncated DcuS"
                     /label=truncated DcuS
                     /protein_id="QKE44343.1"
                     /translation="MTSKTEVRKLMQRLDGLVNYADALRERSHEFMNKLHVILGLLHLK
                     SYKQLEDYILKTANNYQEEIGSLLGKIKSPVIAGFLISKINRATDLGHTLILNSESQLP
                     DSGSEDQVATLITTLGNLIENALEALGPEPGGEISVTLHYRHGWLHCEVNDDGPGIAPD
                     KIDHIFDKGVSTKGSERGVGLALVKQQVENLGGSIAVESEPGIFTQFFVQIPWDGERSN
                     RASASGSTGV"
     misc_feature    1347..1349
                     /label=lysine 324
                     /note="lysine 324"
     misc_feature    1365..1367
                     /label=leu330
                     /note="leu330"
     misc_feature    1377..1379
                     /label=leucin 334
                     /note="leucin 334"
     misc_feature    1389..1391
                     /label=V338
                     /note="V338"
     misc_feature    1392..1394
                     /label=asn339
                     /note="asn339"
     misc_feature    1395..1412
                     /label=alpha helix
                     /note="alpha helix"
     misc_feature    1413..2006
                     /label=Kinase domain 346-538
                     /note="Kinase domain 346-538"
     polyA_signal    2073..2297
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2343..2771
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2785..3114
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3181..3972
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    4149..4282
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4319..4335)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4343..4359)
                     /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(4367..4397)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4412..4433)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4721..5306)
                     /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 gtggtaccct cgaggcgatc tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     1081 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atcgctgact
     1141 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     1201 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaga gctctctggc taactagaga
     1261 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgt
     1321 tcgaagcgga attcaccatg actagtaaaa ctgaagtacg taaactgatg cagcgactcg
     1381 acggtctggt caactatgct gacgcacttc gtgaacgatc ccacgaattt atgaataaat
     1441 tgcatgtgat tctcggatta ttgcatctga agagttataa gcagttggaa gattacattc
     1501 tcaaaacagc caataactat caggaagaga ttggctctct gctgggtaag atcaaatctc
     1561 cggttatcgc tggtttttta atcagcaaga ttaaccgcgc gaccgattta ggccatacgc
     1621 tgattttaaa cagtgaaagc cagctgccag acagcggcag tgaggaccag gtcgcgacgc
     1681 tgattaccac gttgggaaat ctgatagaaa acgcgctgga ggcattaggg ccggaacccg
     1741 gaggcgaaat tagcgtaaca ttgcactacc gtcacggctg gctgcactgt gaagttaatg
     1801 atgatggacc ggggatcgca cccgataaaa tcgatcacat ttttgacaaa ggtgtctcga
     1861 caaaaggaag cgagcgaggc gtcggtttag cacttgtcaa acaacaggta gaaaatctcg
     1921 gcggcagcat cgccgtggaa tcggaacccg ggattttcac acaatttttt gtccagatac
     1981 cctgggacgg ggagaggtcg aacagagcta gtgctagcgg atccaccggt gtctagaaag
     2041 ctttgaggcc ggcctgcagg gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2101 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2161 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2221 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     2281 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     2341 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2401 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2461 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2521 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2581 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2641 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2701 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2761 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     2821 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     2881 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2941 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3001 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     3061 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     3121 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     3181 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3241 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3301 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3361 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3421 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3481 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3541 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     3601 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     3661 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     3721 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     3781 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     3841 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     3901 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     3961 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4021 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4081 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4141 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4201 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4261 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     4321 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4381 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4441 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4501 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4561 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4621 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4681 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4741 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4801 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4861 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4921 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4981 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5041 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5101 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5161 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5221 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga
     5281 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5341 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5401 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5461 aaacttggtc tgacag
//
