LOCUS       Exported                6824 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Retroviral vector pCX4hyg DNA, complete sequence.
ACCESSION   AB086387
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6824)
  AUTHORS   Akagi T, Sasai K, Hanafusa H.
  TITLE     Refractory nature of normal human diploid fibroblasts with respect 
            to oncogene-mediated transformation
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 100 (23), 13567-13572 (2003)
  PUBMED    14597713
REFERENCE   2  (bases 1 to 6824)
  AUTHORS   Akagi T.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-JUN-2002) Tsuyoshi Akagi, Osaka Bioscience Institute, 
            Molecular Oncology; Furuedai 6-2-4, Suita, Osaka 565-0874, Japan 
            (E-mail:takagi@obi.or.jp, Tel:81-6-6872-4834, Fax:81-6-6871-7521)
REFERENCE   3  (bases 1 to 6824)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6824
                     /organism="Retroviral vector pCX4hyg"
                     /mol_type="other DNA"
                     /db_xref="taxon:197853"
     regulatory      1..466
                     /regulatory_class="enhancer"
                     /note="CMV IE enhancer"
     enhancer        88..467
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     repeat_region   467..762
                     /label=R-U5 region of MuLV 5 LTR
                     /note="R-U5 region of MuLV 5 LTR"
     regulatory      763..1667
                     /regulatory_class="other"
                     /note="psi+; extended packaging signal"
     misc_feature    826..1183
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1242..1658
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     misc_feature    1668..2045
                     /label=Mo-MuLV pol region containing splice acceptor
                     /note="Mo-MuLV pol region containing splice acceptor"
     misc_feature    1668..2042
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     misc_feature    2044..2106
                     /label=multiple cloning site
                     /note="multiple cloning site"
     regulatory      2107..2716
                     /regulatory_class="ribosome_binding_site"
                     /note="IRES sequence from ECMV"
     misc_feature    2255..2718
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2742..3776
                     /codon_start=1
                     /gene="hyg"
                     /product="hygromycin-B-phosphotransferase"
                     /label=hyg
                     /protein_id="BAC41728.1"
                     /translation="MDRSGKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVG
                     GRGYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQ
                     DLPETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVY
                     HWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSE
                     AMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSL
                     VDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRP
                     RAKE"
     gene            2742..3776
                     /gene="hyg"
                     /label=hyg
     CDS             2757..3776
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     repeat_region   4119..4710
                     /label=3 LTR of Mo-MuLV
                     /note="3 LTR of Mo-MuLV"
     LTR             4241..4710
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     rep_origin      complement(5011..5599)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5763..6623)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="BAC41729.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(5763..6623)
                     /gene="bla"
                     /label=bla
     promoter        complement(6624..6728)
                     /gene="bla"
                     /label=AmpR promoter