pOPINJ vector (V004227)

LOCUS       40924_33787        6218 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pOPINJ, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6218)
  AUTHORS   Berrow NS, Alderton D, Sainsbury S, Nettleship J, Assenberg R, 
            Rahman N, Stuart DI, Owens RJ.
  TITLE     A versatile ligation-independent cloning method suitable for 
            high-throughput expression screening applications
  JOURNAL   Nucleic Acids Res. 35 (6), E45 (2007)
  PUBMED    17317681
REFERENCE   2  (bases 1 to 6218)
  AUTHORS   Berrow NS.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-JAN-2007) Oxford Protein Production Facility, Oxford 
            University, Roosevelt Drive, Oxford, Oxfordshire OX3 7BN, U.K.
REFERENCE   3  (bases 1 to 6218)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6218)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2007"; volume: "35"; issue: "6"; pages: "E45"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (17-JAN-2007) Oxford Protein Production Facility, Oxford University,
            Roosevelt Drive, Oxford, Oxfordshire OX3 7BN, U.K."
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     181..1008
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     enhancer        1140..1443
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1449..1726
                     /label=chicken beta-actin promoter
     promoter        2150..2168
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2169..2193
                     /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        2209..2318
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     regulatory      2318..2326
                     /regulatory_class="ribosome_binding_site"
     CDS             2339..2356
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             2357..3010
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
     misc_feature    3033..3047
                     /label=5' in-fusion site (after KnpI digest)
                     /note="5' in-fusion site (after KnpI digest)"
     protein_bind    3093..3114
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3129..3159
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3167..3183
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     CDS             3203..3376
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
     misc_feature    3385..3399
                     /label=3' in-fusion site (after HindIII digest)
                     /note="3' in-fusion site (after HindIII digest)"
     CDS             3406..3423
                     /label=6xHis
                     /note="6xHis affinity tag"
     polyA_signal    3578..3633
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     terminator      3721..3768
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     3781..4486
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     protein_bind    complement(4495..4528)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(4696..5283)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5299..6156)
                     /label=AmpR
                     /note="beta-lactamase"