LOCUS       Exported                4529 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFloxin-MCS2-IRES-MCS, complete sequence.
ACCESSION   GU180239
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4529)
  AUTHORS   Singla VI, Julie H, Santos N, Seol A, Norman AR, Wakenight P, 
            Skarnes WC, Reiter JF.
  TITLE     A resource for engineering alleles in mouse embryonic stem cells
  JOURNAL   Nat. Methods (2010) In press
REFERENCE   2  (bases 1 to 4529)
  AUTHORS   Reiter JF.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-NOV-2009) Biochemistry, UCSF, 1550 4th Street, Rock 
            Hall 446, San Francisco, CA 94158, USA
REFERENCE   3  (bases 1 to 4529)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4529
                     /organism="Cloning vector pFloxin-MCS2-IRES-MCS"
                     /mol_type="other DNA"
                     /db_xref="taxon:696211"
     regulatory      1..525
                     /regulatory_class="promoter"
                     /note="b-actin promoter"
     regulatory      520..529
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    532..565
                     /label=lox66
                     /note="lox66"
     protein_bind    532..565
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     intron          566..826
                     /note="Engrailed2 intron"
     misc_feature    827..873
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    874..892
                     /label=tandem stops
                     /note="tandem stops"
     misc_feature    892..1470
                     /label=IRES element
                     /note="IRES element"
     misc_feature    919..1470
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1476..1507
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    1574..1795
                     /label=SV40 polyA region
                     /note="SV40 polyA region"
     polyA_signal    1681..1815
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    1833..1880
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    1834..1880
                     /label=FRT site
                     /note="FRT site"
     primer_bind     complement(1904..1920)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1928..1944
                     /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)."
     promoter        complement(1952..1982)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1997..2018
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2306..2894)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3065..3925)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3926..4030)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4504..4520
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"