pGL3-U6-sgRNA-PGK-puromycin
PVT10975 2ug
pGL3-U6-sgRNA-PGK-puromycin Information
Nuclear resistance: ampicillin Amp
Screening marker: puromycin Puro
Clone strain: Escherichia coli DH5 alpha
Culture conditions: LB medium, 37 C
Remarks: there are differences between sequences of Addgene.
pGL3-U6-sgRNA-PGK-puromycin Description
PGL3-U6-sgRNA-PGK-puromycin is a mammalian gene editing plasmid. U6 promotes the transcription of sgRNA, which can be digested by BsaI and linked to sgRNA sequence. pGK promotes the expression of purinomycin Puro resistance gene.
pGL3-U6-sgRNA-PGK-puromycin Multiple cloning site
pGL3-U6-sgRNA-PGK-puromycin Sequence
LOCUS Exported 4951 bp ds-DNA circular SYN 17-JULY-2017
KEYWORDS pGL3-U6-sgRNA-PGK-puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4951)
AUTHORS
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..4951
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 72..312
/label="U6 Promoter"
/note="U6 promoter"
/note="RNA polymerase III promoter for human U6 snRNA"
misc_RNA 343..418
/note="gRNA scaffold"
/note="guide RNA scaffold for the CRISPR/Cas9 system"
misc_feature 470..587
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 636..1146
/note="hPGK promoter"
/note="human phosphoglycerate kinase 1 promoter"
CDS 1168..1767
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/note="PuroR"
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
polyA_signal 1914..2035
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(2454..3042)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3213..4073)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4074..4178)
/gene="bla"
/note="AmpR promoter"
rep_origin 4205..4660
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
polyA_signal 4791..4839
/note="synthetic polyadenylation signal"
misc_feature 4853..4944
/note="pause site"
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
ORIGIN
1 ggtaccgatt agtgaacgga tctcgacggt atcgatcacg agactagcct cgagcggccg
61 cccccttcac cgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
121 ctgttagaga gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata
181 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
241 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
301 tgtggaaagg acgaaacacc gtgagaccga gagagggtct cagttttaga gctagaaata
361 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
421 tttttaaaga attctcgacc tcgagacaaa tggcagtatt catccacaat tttaaaagaa
481 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
541 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca
601 gggacagcag agatccactt tggccgcggc tcgagggggt tggggttgcg ccttttccaa
661 ggcagccctg ggtttgcgca gggacgcggc tgctctgggc gtggttccgg gaaacgcagc
721 ggcgccgacc ctgggactcg cacattcttc acgtccgttc gcagcgtcac ccggatcttc
781 gccgctaccc ttgtgggccc cccggcgacg cttcctgctc cgcccctaag tcgggaaggt
841 tccttgcggt tcgcggcgtg ccggacgtga caaacggaag ccgcacgtct cactagtacc
901 ctcgcagacg gacagcgcca gggagcaatg gcagcgcgcc gaccgcgatg ggctgtggcc
961 aatagcggct gctcagcagg gcgcgccgag agcagcggcc gggaaggggc ggtgcgggag
1021 gcggggtgtg gggcggtagt gtgggccctg ttcctgcccg cgcggtgttc cgcattctgc
1081 aagcctccgg agcgcacgtc ggcagtcggc tccctcgttg accgaatcac cgacctctct
1141 ccccaggggg atccaccgga gcttaccatg accgagtaca agcccacggt gcgcctcgcc
1201 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
1261 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
1321 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
1381 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
1441 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
1501 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
1561 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
1621 gagcgcgccg gggtgcccgc cttcctggaa acctccgcgc cccgcaacct ccccttctac
1681 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
1741 tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa
1801 aggagcgcac gaccccatgc atcggtacct ttaagaccaa tgacttacaa ggcagctgta
1861 gatcttagcc actttctaga gtcggggcgg ccggccgctt cgagcagaca tgataagata
1921 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
1981 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
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