| 質粒名稱: | pCI H2B-RFP |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 92398 |
| 存儲實驗室: | Tatjana Sauka-Spengler |
|
載體骨架基本信息 |
|
| 載體名稱: | pCI |
| 載體出品公司: | Clontech |
| 空載體大小: | 4935bp |
| 完整質粒大小: | 6600bp |
| 載體類型: | 哺乳細胞表達 |
| 載體抗性: | 氨芐霉素 |
| 生長溫度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷貝數: | 高拷貝 |
|
插入基因信息 |
|
| 插入基因名稱: | IRES H2B RFP |
| 插入大小: | 1665bp |
| 啟動子: | CAG |
| 標簽/融合蛋白: | 人細胞色素c氧化酶亞基VIII（N端） |
| 克隆方法: | Gibson Cloning |
| 5’測序引物 | CGTGCTGGTTATTGTGCTGT |
| 3’測序引物 |
TAAGGAATGGACAGCAGGGG
(Common Sequencing Primers)
|
|
文獻引用方法 |
|
| 材料和方法部分: | pCI H2B-RFP was a gift from Tatjana Sauka-Spengler (Addgene plasmid # 92398 ; //n2t.net/addgene:92398 ; RRID:Addgene_92398) |
| 參考文獻部分: | Genome and epigenome engineering CRISPR toolkit for in vivo modulation of cis-regulatory interactions and gene expression in the chicken embryo. Williams RM, Senanayake U, Artibani M, Taylor G, Wells D, Ahmed AA, Sauka-Spengler T. Development. 2018 Feb 23;145(4). pii: dev.160333. doi: 10.1242/dev.160333. 10.1242/dev.160333 PubMed 29386245 |
買家導航
pCI H2B-RFP載體質粒基本信息
pCI H2B-RFP質粒圖譜載體圖譜和pCI H2B-RFP載體序列質粒序列多克隆位點信息
pCI H2B-RFP質粒載體簡介
pCI H2B-RFP質粒序列載體序列
LOCUS pCI H2B-RFP 6600 bp DNA circular SYN 14-APR-2025
DEFINITION Ubiquitous expression plasmid, contains CAG promoter (CMV immediate
early enhancer and chicken beta actin promoter), MCS and IRES
controlled Histone2B-mRFP1 reporter..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6600)
AUTHORS Williams RM, Senanayake U, Artibani M, Taylor G, Wells D, Ahmed AA,
Sauka-Spengler T
TITLE Genome and epigenome engineering CRISPR toolkit for in vivo
modulation of cis-regulatory interactions and gene expression in the
chicken embryo.
JOURNAL Development. 2018 Feb 23;145(4). pii: dev.160333. doi:
10.1242/dev.160333.
PUBMED 29386245
REFERENCE 2 (bases 1 to 6600)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
//gt12365.cn
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Development."; date: "2018-02-23"; volume: "145(4). pii:
dev.160333. doi"; pages: " 10.1242/dev.160333"
COMMENT Sequence Label: pCI H2B-RFP
FEATURES Location/Qualifiers
source 1..6600
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 5..384
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 386..661
/label=chicken beta-actin promoter
intron 662..1674
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1598..1620
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1682..1701
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
misc_feature join(1835..2409,2410..2412,2413..2421)
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(2002..2019)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 2229..2248
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 2428..2805
/codon_start=1
/gene="HIST1H2BJ"
/product="human histone H2B"
/label=H2B
/translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
GELAKHAVSEGTKAITKYTSAK"
regulatory 2818..2827
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 2824..3501
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
CDS 2824..3501
/codon_start=1
/product="monomeric derivative of DsRed (Campbell et al.,
2002)"
/label=mRFP1
/note="mammalian codon-optimized"
/translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
TGA"
primer_bind complement(2954..2972)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 3222..3241
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 3416..3439
/label=DsRed1-C
/note="DsRed1, forward primer"
primer_bind complement(3592..3611)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 3657..3712
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(3711..3730)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(4103..4119)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4103..4119)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4116..4138)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4127..4143
/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(join(4151..4157,4158..4175,4176..4181))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4196..4217
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4276..4471
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 4322..4457
/label=SV40 ori
/note="SV40 origin of replication"
polyA_signal 4477..4611
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4514..4533)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4568..4587
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4679..4696)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4850..5438)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4930..4949)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(5609..6400,6401..6469))
/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 6232..6251
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6470..6574)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc
1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
1621 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattgct cgagagttcg
1741 aagccttaag tacgcgtagg cgcgccttaa ttaacgttta aacgctagct catcgataag
1801 cttgatatcg aattcctgca gcccggggga tccgcccctc tccctccccc ccccctaacg
1861 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
1921 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
1981 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
2041 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
2101 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
2161 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
2221 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
2281 cccattgtat gggatctgat ctggggcctc ggtacacatg ctttacatgt gtttagtcga
2341 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
2401 gatgataata tggccacaac catggttatg ccagagccag cgaagtctgc tcccgccccg
2461 aaaaagggct ccaagaaggc ggtgactaag gcgcagaaga aaggcggcaa gaagcgcaag
2521 cgcagccgca aggagagcta ttccatctat gtgtacaagg ttctgaagca ggtccaccct
2581 gacaccggca tttcgtccaa ggccatgggc atcatgaatt cgtttgtgaa cgacattttc
2641 gagcgcatcg caggtgaggc ttcccgcctg gcgcattaca acaagcgctc gaccatcacc
2701 tccagggaga tccagacggc cgtgcgcctg ctgctgcctg gggagttggc caagcacgcc
2761 gtgtccgagg gtactaaggc catcaccaag tacaccagcg ctaaggatcc accggtcgcc
2821 accatggcct cctccgagga cgtcatcaag gagttcatgc gcttcaaggt gcgcatggag
2881 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
2941 ggcacccaga ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac
3001 atcctgtccc ctcagttcca gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
3061 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
3121 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
3181 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
3241 accatgggct gggaggcctc caccgagcgg atgtaccccg aggacggcgc cctgaagggc
3301 gagatcaaga tgaggctgaa gctgaaggac ggcggccact acgacgccga ggtcaagacc
3361 acctacatgg ccaagaagcc cgtgcagctg cccggcgcct acaagaccga catcaagctg
3421 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgagcg cgccgagggc
3481 cgccactcca ccggcgccta agcggccgca attcactcct caggtgcagg ctgcctatca
3541 gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc
3601 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata
3661 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
3721 gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc
3781 aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta
3841 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta
3901 gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt
3961 acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc
4021 tcttctctta tggagatccc tcgacctgca ccgtcgacca gctggtcgac ggtgcaccgt
4081 cgaccagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
4141 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
4201 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
4261 cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
4321 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
4381 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
4441 aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat
4501 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
4561 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt
4621 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
4681 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
4741 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
4801 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
4861 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
4921 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
4981 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
5041 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
5101 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
5161 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
5221 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
5281 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
5341 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
5401 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
5461 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
5521 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
5581 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
5641 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
5701 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
5761 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
5821 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
5881 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
5941 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
6001 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
6061 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
6121 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
6181 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
6241 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
6301 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
6361 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
6421 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
6481 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
6541 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//