pEFBOS-creIRESPuro vector (V007607)

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LOCUS       40924_17089        8102 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pEFBOS-creIRESPuro, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8102)
  AUTHORS   Davis RP, Ng ES, Costa M, Mossman AK, Sourris K, Elefanty AG, 
            Stanley EG.
  TITLE     Targeting a GFP reporter gene to the MIXL1 locus of human embryonic 
            stem cells identifies human primitive streak-like cells and enables 
            isolation of primitive hematopoietic precursors
  JOURNAL   Blood 111 (4), 1876-1884 (2008)
  PUBMED    18032708
REFERENCE   2  (bases 1 to 8102)
  AUTHORS   Davis RP, Costa M, Grandela C, Holland AM, Hatzistavrou T, Micallef 
            SJ, Li X, Goulburn AL, Azzola L, Elefanty AG, Stanley EG.
  TITLE     A protocol for removal of antibiotic resistance cassettes from human
            embryonic stem cells genetically modified by homologous 
            recombination or transgenesis
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 8102)
  AUTHORS   Davis RP, Stanley EG, Elefanty AG.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-MAY-2008) MISCL, Monash University, Wellington, 
            Clayton, Victoria 3800, Australia
REFERENCE   4  (bases 1 to 8102)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 8102)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Blood"; 
            date: "2008"; volume: "111"; issue: "4"; pages: "1876-1884"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (02-MAY-2008) MISCL, Monash University, Wellington, Clayton, 
            Victoria 3800, Australia"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8102
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(333..543)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        562..1743
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             1766..1786
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1784..2812
                     /codon_start=1
                     /label=Cre
                     /note="site-specific recombinase"
                     /translation="VSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     misc_feature    2849..3435
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             3445..4041
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="LTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    4125..4349
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     3'UTR           4522..5222
                     /label=bovine growth hormone 3' UTR
                     /note="bovine growth hormone 3' UTR"
     primer_bind     complement(5231..5247)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5460..5915
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6197..6301
                     /label=AmpR promoter
     CDS             6302..7159
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7333..7921
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"