LOCUS       Exported               10495 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Retroviral vector that expresses wild type Flag-tagged human USP7.
ACCESSION   .
VERSION     .
KEYWORDS    pQFlag-USP7 WT puroR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10495)
  AUTHORS   Maertens GN, El Messaoudi-Aubert S, Elderkin S, Hiom K, Peters G
  TITLE     Ubiquitin-specific proteases 7 and 11 modulate Polycomb regulation 
            of the INK4a tumour suppressor.
  JOURNAL   EMBO J. 2010 Aug 4;29(15):2553-65. doi: 10.1038/emboj.2010.129. Epub
            2010 Jul 2.
  PUBMED    20601937
REFERENCE   2  (bases 1 to 10495)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/emboj.2010.129"; journalName: "EMBO J"; date: "2010-08-4- 
            4"; volume: "29"; issue: "15"; pages: "2553-65"
FEATURES             Location/Qualifiers
     source          1..10495
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        309..512
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             513..688
                     /label=5 LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    751..950
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1151..1567
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1497..1519
                     /label=pLXSN 5
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1535..1551
                     /label=pBABE 5
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1604..1907
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1908..2111
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2108..2132
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2268..2291
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    5622..6195
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5789..5806)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6016..6035
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6228..6827
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6228..6247)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6724..6744
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             7201..7626
                     /label=3 LTR (Delta-U3)
                     /note="self-inactivating 3 long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(7729..7751)
                     /label=pGEX 3
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7887..7907)
                     /label=pBABE 3
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7892..8221
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8072..8207
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8134..8153
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     8243..8262
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(8343..8360)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8514..9102)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8594..8613)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9273..10133)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     9896..9915
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10134..10238)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     10306..10324
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(10424..10495,1..308)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"