pLeo1246 vector (V016121)

LOCUS       62056_13990        5848 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pLeo1246, complete sequence.
ACCESSION   MT267307
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5848)
  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 5848)
  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 5848)
  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..5848
                     /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"
     misc_feature    1335..1340
                     /label=Kozak+ATG
                     /note="Kozak+ATG"
     CDS             1338..2408
                     /codon_start=1
                     /transl_table=11
                     /product="ORK"
                     /label=ORK
                     /protein_id="QKE44346.1"
                     /translation="MTSGSQVQLVESGGGLVQAGGSLRLSCTASGRTGTIYSMAWFRQA
                     PGKEREFLATVGWSSGITYYMDSVKGRFTISRDKGKNTVYLQMDSLKPEDTAVYYCTAT
                     RAYSVGYDYWGQGTQVTVSSASGGGGSASLMQRLDGLVNYADALRERSHEFMNKLHVIL
                     GLLHLKSYKQLEDYILKTANNYQEEIGSLLGKIKSPVIAGFLISKINRATDLGHTLILN
                     SESQLPDSGSEDQVATLITTLGNLIENALEALGPEPGGEISVTLHYRHGWLHCEVNDDG
                     PGIAPDKIDHIFDKGVSTKGSERGVGLALVKQQVENLGGSIAVESEPGIFTQFFVQIPW
                     DGERSNRASASGSTGV"
     misc_feature    1347..1709
                     /label=aCaffVHH
                     /note="aCaffVHH"
     misc_feature    1428..1451
                     /label=CDR1
                     /note="CDR1"
     misc_feature    1500..1523
                     /label=CDR2
                     /note="CDR2"
     misc_feature    1641..1676
                     /label=CDR3
                     /note="CDR3"
     misc_feature    1716..1730
                     /label=scfv
                     /note="scfv"
     misc_feature    1737..2378
                     /label=DcuS truncation
                     /note="DcuS truncation"
     misc_feature    1737..1739
                     /label=leu330
                     /note="leu330"
     misc_feature    1749..1751
                     /label=leucin 334
                     /note="leucin 334"
     misc_feature    1761..1763
                     /label=V338
                     /note="V338"
     misc_feature    1764..1766
                     /label=asn339
                     /note="asn339"
     misc_feature    1767..1784
                     /label=alpha helix
                     /note="alpha helix"
     misc_feature    1785..2378
                     /label=Kinase domain 346-538
                     /note="Kinase domain 346-538"
     polyA_signal    2445..2669
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2715..3143
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3157..3486
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3553..4344
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    4521..4654
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4691..4707)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4715..4731)
                     /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(4739..4769)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4784..4805)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5093..5678)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"