| 质粒名称: | pEGFP-puro |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 45561 |
| 存储实验室: | Michael McVoy |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-C1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4731 bp |
| 完整质粒大小: | 5310 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达；双功能选择蛋白 |
| 筛选标记: | 嘌呤霉素；EGFP |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Puromycin N-acetyl transferase,PuroR,EGFP-Puro |
| 物种来源: | -- |
| 插入大小: | 600 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | Phcmv (Strong human cytomegalovirus immediate early promoter) |
| 标签/融合蛋白: | EGFP（主干网上的N端子） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BglII(未破坏) |
| 3’克隆位点: | BamHI(未破坏) |
| 5’测序引物 | EGFP-C; CMV-F |
| 3’测序引物 | SV40-pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pEGFP-puro was a gift from Michael McVoy (Addgene plasmid # 45561 ; http://n2t.net/addgene:45561 ; RRID:Addgene_45561) |
| 参考文献部分: | Bifunctional protein conferring enhanced green fluorescence and puromycin resistance. Abbate J, Lacayo JC, Prichard M, Pari G, McVoy MA. Biotechniques. 2001 Aug;31(2):336-40. 10.2144/01312st05 PubMed 11515370 |
买家导航
pEGFPpuro载体基本信息
pEGFPpuro载体图谱质粒图谱和多克隆位点信息
pEGFPpuro载体简介
pEGFPpuro载体序列
LOCUS pEGFP-puro 5167 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pEGFP-puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5167)
AUTHORS Abbate J, Lacayo JC, Prichard M, Pari G, McVoy MA
TITLE Bifunctional protein conferring enhanced green fluorescence and
puromycin resistance.
JOURNAL Biotechniques. 2001 Aug;31(2):336-40.
PUBMED 11515370
REFERENCE 2 (bases 1 to 5167)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5167
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 71..374
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 375..578
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 529..549
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 617..626
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 623..1339
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 623..1339
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(668..689)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(929..948)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1276..1297
/label=EGFP-C
/note="EGFP, forward primer"
CDS 1370..1963
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPG"
primer_bind complement(1370..1389)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 1866..1886
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 2096..2217
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2133..2152)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2187..2206
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(2234..2250)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2235..2251)
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind complement(2275..2294)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(2276..2294)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2304..2321)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(2304..2320)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2313..2335)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin complement(2461..2917)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2599..2620)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2811..2830
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2944..3048
/gene="bla"
/label=AmpR promoter
CDS 3049..3909
/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 complement(3267..3286)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4080..4668
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4569..4588
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4822..4839
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 4956..4977
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4992..5022
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5030..5046
/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)."
primer_bind 5035..5057
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 5054..5070
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5054..5070
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 5089..5109
/label=T3
/note="T3 promoter, forward primer"
promoter 5091..5109
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind 5146..5162
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5146..5162
/label=pBluescriptSK
/note="For pBluescript vector"
ORIGIN
1 ggctgcagga attcgattaa tagtaatcaa ttacggggtc attagttcat agcccatata
61 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
121 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
181 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
241 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
301 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
361 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
421 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
481 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
541 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
601 gctagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
661 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
721 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
781 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
841 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
901 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
961 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
1021 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
1081 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
1141 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
1201 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
1261 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
1321 catggacgag ctgtacaagt ccggactcag atctaggaga cgaccttcca tgaccgagta
1381 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc
1441 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga
1501 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
1561 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
1621 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
1681 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
1741 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
1801 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
1861 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
1921 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcgggat ccaccggatc
1981 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
2041 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
2101 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
2161 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg
2221 cgatcaagct tatcgatacc gtcgacctcg agggggggcc cggtacccaa ttcgccctat
2281 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
2341 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
2401 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
2461 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
2521 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
2581 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
2641 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
2701 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
2761 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
2821 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
2881 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
2941 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
3001 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
3061 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
3121 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
3181 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
3241 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
3301 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
3361 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
3421 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
3481 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
3541 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
3601 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
3661 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
3721 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
3781 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
3841 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
3901 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
3961 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
4021 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
4081 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
4141 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
4201 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
4261 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
4321 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
4381 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
4441 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
4501 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
4561 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
4621 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
4681 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
4741 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
4801 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
4861 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
4921 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
4981 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
5041 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct
5101 cactaaaggg aacaaaagct ggagctccac cgcggtggcg gccgctctag aactagtgga
5161 tcccccg
//
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