pAAVS1-Puro-DNR vector (V012309)

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LOCUS       pAAVS1-Puro-DNR.        7507 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Mammalian donor vector for transgene integration at the AAVS1 locus 
            in the human genome. Use together with pCas-Guide-AAVS1.
ACCESSION   .
VERSION     .
KEYWORDS    pAAVS1-Puro-DNR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7507)
  AUTHORS   OriGene
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7507)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(119..135)
                     /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)."
     promoter        complement(143..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(188..209)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(497..1085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1259..2116)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(2117..2221)
                     /label=AmpR promoter
     rep_origin      complement(2503..2958)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     3171..3187
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3278..3779
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     protein_bind    complement(3792..3824)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    complement(3851..4304)
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation
                     signal"
     CDS             complement(4384..4980)
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     promoter        complement(5020..5519)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     enhancer        5637..5940
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5941..6144
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        6170..6188
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    6197..6300
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             6303..6332
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     CDS             6351..6374
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     primer_bind     complement(6394..6410)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    6421..6897
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     misc_feature    6972..7501
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"