LOCUS pCMVHA hEZH2 8805 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Mammalian expression of human EZH2 with HA tag.
ACCESSION .
VERSION .
KEYWORDS pCMVHA hEZH2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8805)
AUTHORS Bracken AP, Pasini D, Capra M, Prosperini E, Colli E, Helin K
TITLE EZH2 is downstream of the pRB-E2F pathway, essential for
proliferation and amplified in cancer.
JOURNAL EMBO J. 2003 Oct 15. 22(20):5323-35.
PUBMED 14532106
REFERENCE 2 (bases 1 to 8805)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8805
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 198..577
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 578..781
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 732..752
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 778..802
/label=LNCX
/note="Human CMV promoter, forward primer"
intron 893..1465
/label=-beta-globin intron
/note="intron from rabbit beta-globin gene"
CDS 1545..1571
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
primer_bind complement(3924..3943)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 3989..4044
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(4043..4062)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(4402..4418)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4402..4418)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4415..4437)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4426..4442
/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(4450..4480)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4495..4516
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4633..4650)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4804..5392)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4884..4903)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5563..6423)
/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"
primer_bind 6186..6205
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6424..6528)
/gene="bla"
/label=AmpR promoter
primer_bind 6596..6614
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(6652..6674)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 6774..6793
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6987..7009
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 7001..7018
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 7002..7018
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 7429..7574
/label=HSV TK promoter
/note="herpes simplex virus thymidine kinase promoter"
CDS 7631..8425
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(7685..7704)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 8295..8314
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(8619..8638)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 8663..8710
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
1 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaagc
61 cctgccatag ccacgggccc cgtgggttag ggacggcgga tcgcggccct ctagagagct
121 tggcccattg catacgttgt atccatatca taatatgtac atttatattg gctcatgtcc
181 aacattaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
241 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
301 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
361 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
421 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
481 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
541 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
601 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
661 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
721 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
781 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
841 aagacaccgg gaccgatcca gcctccggtc gaccgatcct gagaacttca gggtgagttt
901 ggggaccctt gattgttctt tctttttcgc tattgtaaaa ttcatgttat atggaggggg
961 caaagttttc agggtgttgt ttagaatggg aagatgtccc ttgtatcacc atggaccctc
1021 atgataattt tgtttctttc actttctact ctgttgacaa ccattgtctc ctcttatttt
1081 cttttcattt tctgtaactt tttcgttaaa ctttagcttg catttgtaac gaatttttaa
1141 attcactttt gtttatttgt cagattgtaa gtactttctc taatcacttt tttttcaagg
1201 caatcagggt atattatatt gtacttcagc acagttttag agaacaattg ttataattaa
1261 atgataaggt agaatatttc tgcatataaa ttctggctgg cgtggaaata ttcttattgg
1321 tagaaacaac tacaccctgg tcatcatcct gcctttctct ttatggttac aatgatatac
1381 actgtttgag atgaggataa aatactctga gtccaaaccg ggcccctctg ctaaccatgt
1441 tcatgccttc ttctctttcc tacagctcct gggcaacgtg ctggttgttg tgctgtctca
1501 tcattttggc aaagaattcc tcgacggatc aattcaccat ggcataccca tacgatgttc
1561 cagattacgc tagcggatcc atgggccaga ctgggaagaa atctgagaag ggaccagttt
1621 gttggcggaa gcgtgtaaaa tcagagtaca tgcgactgag acagctcaag aggttcagac
1681 gagctgatga agtaaagagt atgtttagtt ccaatcgtca gaaaattttg gaaagaacgg
1741 aaatcttaaa ccaagaatgg aaacagcgaa ggatacagcc tgtgcacatc ctgacttctg
1801 tgagctcatt gcgcgggact agggagtgtt cggtgaccag tgacttggat tttccaacac
1861 aagtcatccc attaaagact ctgaatgcag ttgcttcagt acccataatg tattcttggt
1921 ctcccctaca gcagaatttt atggtggaag atgaaactgt tttacataac attccttata
1981 tgggagatga agttttagat caggatggta ctttcattga agaactaata aaaaattatg
2041 atgggaaagt acacggggat agagaatgtg ggtttataaa tgatgaaatt tttgtggagt
2101 tggtgaatgc ccttggtcaa tataatgatg atgacgatga tgatgatgga gacgatcctg
2161 aagaaagaga agaaaagcag aaagatctgg aggatcaccg agatgataaa gaaagccgcc
2221 cacctcggaa atttccttct gataaaattt ttgaagccat ttcctcaatg tttccagata
2281 agggcacagc agaagaacta aaggaaaaat ataaagaact caccgaacag cagctcccag
2341 gcgcacttcc tcctgaatgt acccccaaca tagatggacc aaatgctaaa tctgttcaga
2401 gagagcaaag cttacactcc tttcatacgc ttttctgtag gcgatgtttt aaatatgact
2461 gcttcctaca tccttttcat gcaacaccca acacttataa gcggaagaac acagaaacag
2521 ctctagacaa caaaccttgt ggaccacagt gttaccagca tttggaggga gcaaaggagt
2581 ttgctgctgc tctcaccgct gagcggataa agaccccacc aaaacgtcca ggaggccgca
2641 gaagaggacg gcttcccaat aacagtagca ggcccagcac ccccaccatt aatgtgctgg
2701 aatcaaagga tacagacagt gatagggaag cagggactga aacgggggga gagaacaatg
2761 ataaagaaga agaagagaag aaagatgaaa cttcgagctc ctctgaagca aattctcggt
2821 gtcaaacacc aataaagatg aagccaaata ttgaacctcc tgagaatgtg gagtggagtg
2881 gtgctgaagc ctcaatgttt agagtcctca ttggcactta ctatgacaat ttctgtgcca
2941 ttgctaggtt aattgggacc aaaacatgta gacaggtgta tgagtttaga gtcaaagaat
3001 ctagcatcat agctccagct cccgctgagg atgtggatac tcctccaagg aaaaagaaga
3061 ggaaacaccg gttgtgggct gcacactgca gaaagataca gctgaaaaag gacggctcct
3121 ctaaccatgt ttacaactat caaccctgtg atcatccacg gcagccttgt gacagttcgt
3181 gcccttgtgt gatagcacaa aatttttgtg aaaagttttg tcaatgtagt tcagagtgtc
3241 aaaaccgctt tccgggatgc cgctgcaaag cacagtgcaa caccaagcag tgcccgtgct
3301 acctggctgt ccgagagtgt gaccctgacc tctgtcttac ttgtggagcc gctgaccatt
3361 gggacagtaa aaatgtgtcc tgcaagaact gcagtattca gcggggctcc aaaaagcatc
3421 tattgctggc accatctgac gtggcaggct gggggatttt tatcaaagat cctgtgcaga
3481 aaaatgaatt catctcagaa tactgtggag agattatttc tcaagatgaa gctgacagaa
3541 gagggaaagt gtatgataaa tacatgtgca gctttctgtt caacttgaac aatgattttg
3601 tggtggatgc aacccgcaag ggtaacaaaa ttcgttttgc aaatcattcg gtaaatccaa
3661 actgctatgc aaaagttatg atggttaacg gtgatcacag gataggtatt tttgccaaga
3721 gagccatcca gactggcgaa gagctgtttt ttgattacag atacagccag gctgatgccc
3781 tgaagtatgt cggcatcgaa agagaaatgg aaatcccttg aactagttgg atccgtcgag
3841 gaattcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc
3901 ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg
3961 aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg
4021 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa
4081 catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg
4141 aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc
4201 ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta
4261 tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc
4321 ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc cctcgacgga
4381 tcggccgcaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac
4441 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
4501 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
4561 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
4621 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
4681 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
4741 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
4801 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
4861 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
4921 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
4981 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
5041 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
5101 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
5161 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
5221 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
5281 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
5341 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
5401 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
5461 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
5521 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
5581 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
5641 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
5701 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
5761 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
5821 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
5881 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
5941 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
6001 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
6061 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
6121 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
6181 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
6241 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
6301 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
6361 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
6421 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
6481 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
6541 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
6601 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
6661 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
6721 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
6781 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
6841 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
6901 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
6961 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc
7021 caagcttcac gctgccgcaa gcactcaggg cgcaagggct gctaaaggaa gcggaacacg
7081 tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta ctgggctatc
7141 tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg
7201 cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc agctggggcg
7261 ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaagg
7321 atctgatggc gcaggggatc aagatcctgc ttcatccccg tggcccgttg ctcgcgtttg
7381 ctggcggtgt ccccggaaga aatatatttg catgtcttta gttctatgat gacacaaacc
7441 ccgcccagcg tcttgtcatt ggcgaattcg aacacgcaga tgcagtcggg gcggcgcggt
7501 cccaggtcca cttcgcatat taaggtgacg cgtgtggcct cgaacaccga gcgaccctgc
7561 agcgacccgc ttaacagcgt caacagcgtg ccgcagatct gatcaagaga caggatgagg
7621 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
7681 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
7741 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
7801 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
7861 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
7921 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
7981 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
8041 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
8101 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
8161 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
8221 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
8281 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
8341 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
8401 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
8461 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
8521 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
8581 gagttcttcg cccaccccgg gagatggggg aggctaactg aaacacggaa ggagacaata
8641 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
8701 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
8761 agaccccatt ggggccaata cgcccgcgtt tcttcctttt cccca
//
