pLeo1248 vector (V016299)

LOCUS       62056_14000        5818 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pLeo1248, complete sequence.
ACCESSION   MT267309
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5818)
  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 5818)
  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 5818)
  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..5818
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..9
                     /label=SV40 polyA
                     /note="SV40 polyA"
     primer_bind     complement(49..65)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(73..89)
                     /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(97..127)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(142..163)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(451..1036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1210..2067)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(2068..2172)
                     /label=AmpR promoter
     misc_feature    2241..2444
                     /label=SV40 early promoter
                     /note="SV40 early promoter"
     promoter        2494..2512
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2541..2546
                     /label=Kozak+ATG
                     /note="Kozak+ATG"
     CDS             2544..3584
                     /codon_start=1
                     /transl_table=11
                     /product="ORK"
                     /label=ORK
                     /protein_id="QKE44348.1"
                     /translation="MTSGSQVQLVESGGGLVQAGGSLRLSCTASGRTGTIYSMAWFRQA
                     PGKEREFLATVGWSSGITYYMDSVKGRFTISRDKGKNTVYLQMDSLKPEDTAVYYCTAT
                     RAYSVGYDYWGQGTQVTVSSASGGGGSASYADALRERSHEFMNKLHVILGLLHLKSYKQ
                     LEDYILKTANNYQEEIGSLLGKIKSPVIAGFLISKINRATDLGHTLILNSESQLPDSGS
                     EDQVATLITTLGNLIENALEALGPEPGGEISVTLHYRHGWLHCEVNDDGPGIAPDKIDH
                     IFDKGVSTKGSERGVGLALVKQQVENLGGSIAVESEPGIFTQFFVQIPWDGERSNRASA
                     SGSTGV"
     misc_feature    2553..2915
                     /label=aCaffVHH
                     /note="aCaffVHH"
     misc_feature    2634..2657
                     /label=CDR1
                     /note="CDR1"
     misc_feature    2706..2729
                     /label=CDR2
                     /note="CDR2"
     misc_feature    2847..2882
                     /label=CDR3
                     /note="CDR3"
     misc_feature    2922..2936
                     /label=scfv
                     /note="scfv"
     misc_feature    2943..3554
                     /label=DcuS truncation
                     /note="DcuS truncation"
     misc_feature    2943..2960
                     /label=alpha helix
                     /note="alpha helix"
     misc_feature    2943..2945
                     /label=Y334
                     /note="Y334"
     misc_feature    2961..3554
                     /label=Kinase domain 346-538
                     /note="Kinase domain 346-538"
     polyA_signal    3621..3845
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3891..4319
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4333..4662
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4729..5520
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    5697..5818
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"