pSiRPG vector (V003176)

LOCUS       40924_40472        8438 bp DNA     circular SYN 18-DEC-2018
DEFINITION  ShRNA expression vector pSiRPG, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8438)
  AUTHORS   Storvold GL, Gjernes E, Askautrud HA, Borresen-Dale AL, Perou CM, 
            Frengen E.
  TITLE     A retroviral vector for siRNA expression in mammalian cells
  JOURNAL   Mol. Biotechnol. 35 (3), 275-282 (2007)
  PUBMED    17652791
REFERENCE   2  (bases 1 to 8438)
  AUTHORS   Stoervold GL.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-MAR-2006) Institute for Medical Genetics, University 
            of Oslo, Norway, PO Box 1036 Blindern, Oslo N-0315, Norway
REFERENCE   3  (bases 1 to 8438)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8438)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. 
            Biotechnol."; date: "2007"; volume: "35"; issue: "3"; pages: 
            "275-282"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-MAR-2006) Institute for Medical Genetics, University of Oslo, 
            Norway, PO Box 1036 Blindern, Oslo N-0315, Norway"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    211..229
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     enhancer        450..829
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        830..1033
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        1078..1096
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1218..1234)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1253..1669)
                     /codon_start=1
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
                     /translation="GQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTF
                     NVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKP
                     PPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA"
     misc_feature    complement(1736..2077)
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell
                     virus"
     LTR             complement(2141..2657)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem
                     cell virus"
     promoter        3485..3589
                     /label=AmpR promoter
     CDS             3590..4447
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4621..5209
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5497..5518
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5533..5563
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5571..5587
                     /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)."
     repeat_region   5750..6103
                     /label=3' LTR
                     /note="3' LTR"
     CDS             complement(6176..6772)
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     promoter        complement(6782..7111)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             complement(7188..7904)
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     promoter        complement(7939..8438)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"