LOCUS       Exported                5811 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCAG.I-SceI, complete sequence.
ACCESSION   JF714900
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5811)
  AUTHORS   Holkers M, de Vries AA, Goncalves MA.
  TITLE     Nonspaced inverted DNA repeats are preferential targets for 
            homology-directed gene repair in mammalian cells
  JOURNAL   Nucleic Acids Res. 40 (5), 1984-1999 (2012)
  PUBMED    22080552
REFERENCE   2  (bases 1 to 5811)
  AUTHORS   Holkers M, de Vries AF, Goncalves MAF.V.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-MAR-2011) Molecular Cell Biology, Leiden University 
            Medical Center, Einthovenweg 20, Leiden, Zuid-Holland 2333 ZC, The 
            Netherlands
REFERENCE   3  (bases 1 to 5811)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5811
                     /organism="Cloning vector pCAG.I-SceI"
                     /mol_type="other DNA"
                     /db_xref="taxon:1109021"
     regulatory      1..381
                     /regulatory_class="enhancer"
                     /note="cytomegalovirus immediate-early gene enhancer"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     regulatory      382..1637
                     /regulatory_class="promoter"
                     /note="chicken beta-actin gene promoter"
     promoter        383..658
                     /label=chicken beta-actin promoter
     intron          659..1667
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     CDS             1639..2580
                     /codon_start=1
                     /product="I-SceI"
                     /label=I-SceI
                     /note="homing endonuclease; meganuclease derived from 
                     Saccharomyces cerevisae"
                     /protein_id="AET06003.1"
                     /translation="MFMPSSFSYSSWATCWLLCCLIILAKNSSKFRSAATMGSRSPKKK
                     RKVPKKHAAPPKKKRKVEDPRFMYPYDVPDYAGMKNIKKNQVMNLGPNSKLLKEYKSQL
                     IELNIEQFEAGIGLILGDAYIRSRDEGKTYCMQFEWKNKAYMDHVCLLYDQWVLSPPHK
                     KERVNHLGNLVITWGAQTFKHQAFNKLANLFIVNNKKTIPNNLVENYLTPMSLAYWFMD
                     DGGKWDYNKNSTNKSIVLNTQSFTFEEVEYLVKGLRNKFQLNCYVKINKNKPIIYIDSM
                     SYLIFYNLIKPYLIPQMMYKLPNTISSETFLK"
     CDS             1762..1782
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1804..1824
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1843..1869
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2594..3121
                     /label=rabbit beta-globin polyadenylation signal
                     /note="rabbit beta-globin polyadenylation signal"
     polyA_site      2742..2797
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(3159..3175)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3183..3199
                     /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(3207..3237)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3252..3273
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3561..4149)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4320..5180)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="ampicillin antibiotic resistance"
                     /protein_id="AET06002.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5181..5285)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5759..5775
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"