LOCUS       62056_14010        5866 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pLeo1254, complete sequence.
ACCESSION   MT267311
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"
     CDS             complement(4..861)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(862..966)
                     /label=AmpR promoter
     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..2426
                     /codon_start=1
                     /transl_table=11
                     /product="C-ORK"
                     /label=C-ORK
                     /protein_id="QKE44350.1"
                     /translation="MTSKTEVRKLMQRLDGLVNYADALRERSHEFMNKLHVILGLLHLK
                     SYKQLEDYILKTANNYQEEIGSLLGKIKSPVIAGFLISKINRATDLGHTLILNSESQLP
                     DSGSEDQVATLITTLGNLIENALEALGPEPGGEISVTLHYRHGWLHCEVNDDGPGIAPD
                     KIDHIFDKGVSTKGSERGVGLALVKQQVENLGGSIAVESEPGIFTQFFVQIPWDGERSN
                     RASASGGGGSASGSQVQLVESGGGLVQAGGSLRLSCTASGRTGTIYSMAWFRQAPGKER
                     EFLATVGWSSGITYYMDSVKGRFTISRDKGKNTVYLQMDSLKPEDTAVYYCTATRAYSV
                     GYDYWGQGTQVTVSSASGSTGV"
     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"
     misc_feature    2034..2402
                     /label=aCaffVHH
                     /note="aCaffVHH"
     misc_feature    2121..2144
                     /label=CDR1
                     /note="CDR1"
     misc_feature    2193..2216
                     /label=CDR2
                     /note="CDR2"
     misc_feature    2334..2369
                     /label=CDR3
                     /note="CDR3"
     polyA_signal    2463..2687
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2733..3161
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3175..3504
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3571..4362
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    4539..4672
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4709..4725)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4733..4749)
                     /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(4757..4787)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4802..4823)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5111..5696)
                     /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 gtgctagtgg cggcggcggc agtgctagtg
     2041 gctctcaggt tcaattggtg gaatctggag ggggtctcgt acaggcaggc ggttctctcc
     2101 gactgagttg cacagcctcc ggtaggactg ggaccatcta ctcaatggcc tggtttcgcc
     2161 aggccccagg caaagaaaga gagtttcttg ccactgtagg ttggagttct gggatcacat
     2221 actacatgga ttcagttaaa ggaagattca ctatcagccg agataaaggg aaaaatactg
     2281 tgtacctcca gatggactct ctgaaaccgg aggacacggc tgtctactac tgtacagcca
     2341 cccgcgccta ctccgtaggg tatgactact gggggcaagg aacacaggta accgtctcta
     2401 gcgctagcgg atccaccggt gtctagaaag ctttgaggcc ggcctgcagg gatcagcctc
     2461 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2521 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2581 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2641 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2701 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2761 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2821 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2881 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2941 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3001 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3061 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3121 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3181 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3241 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3301 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3361 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3421 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3481 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3541 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3601 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3661 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3721 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3781 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3841 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3901 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3961 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4021 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4081 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4141 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4201 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4261 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4321 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4381 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4441 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4501 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4561 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4621 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4681 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4741 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4801 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4861 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4921 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4981 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5041 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5101 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5161 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5221 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5281 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5341 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5401 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5461 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5521 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5581 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5641 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5701 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5761 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5821 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacag
//
