| 质粒名称: | pCI-6.20 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 41002 |
| 存储实验室: | James Inglese |
|
载体骨架基本信息 |
|
| 载体名称: | pGL3-Control |
| 载体出品公司: | Promega |
| 空载体大小: | 5256 bp |
| 完整质粒大小: | 6264bp |
| 载体类型: | 哺乳细胞表达, Luciferase |
| 载体抗性: | 氨苄霉素 |
| 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: | GSG-P2A-RLuc |
| 插入大小: | 1014bp |
| 启动子: | SV40 |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRI (not destroyed) |
| 3’克隆位点: | EcoRI (not destroyed) |
| 5’测序引物 | SV40pro-F |
| 3’测序引物 | EBV-Rev |
|
文献引用方法 |
|
| 材料和方法部分: | pCI-6.20 was a gift from James Inglese (Addgene plasmid # 41002 ; http://n2t.net/addgene:41002 ; RRID:Addgene_41002) |
| 参考文献部分: | A coincidence reporter-gene system for high-throughput screening. Cheng KC, Inglese J. Nat Methods. 2012 Oct;9(10):937. doi: 10.1038/nmeth.2170. 10.1038/nmeth.2170 PubMed 23018994 |
买家导航
pCI-6.20载体质粒基本信息
pCI-6.20质粒图谱载体图谱和pCI-6.20载体序列质粒序列多克隆位点信息
pCI-6.20质粒载体简介
pCI-6.20质粒序列载体序列
LOCUS pCI-6.20 6264 bp DNA circular SYN 14-APR-2025
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6264)
AUTHORS Cheng KC, Inglese J
TITLE A coincidence reporter-gene system for high-throughput screening.
JOURNAL Nat Methods. 2012 Oct;9(10):937. doi: 10.1038/nmeth.2170.
PUBMED 23018994
REFERENCE 2 (bases 1 to 6264)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Methods."; date: "2012-10"; volume: "9(10):937. doi"; pages: "
10.1038/nmeth.2170"
COMMENT Sequence Label: pCI-6.20
FEATURES Location/Qualifiers
source 1..6264
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 48..244
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 95..230
/label=SV40 ori
/note="SV40 origin of replication"
regulatory 274..283
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 280..1929
/codon_start=1
/gene="luc+"
/product="firefly luciferase"
/label=luciferase
/note="enhanced luc+ version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
primer_bind complement(346..364)
/label=LucNrev
/note="Firefly luciferase, reverse primer"
CDS 1945..2001
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 2002..2937
/codon_start=1
/product="luciferase from the anthozoan coelenterate
Renilla reniformis (sea pansy)"
/label=Rluc
/translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
polyA_signal 2981..3102
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2992..3011)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 3046..3065
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3424..3444
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
primer_bind complement(3596..3613)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3767..4355)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3847..3866)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(4526..5317,5318..5386))
/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 5149..5168
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5387..5491)
/gene="bla"
/label=AmpR promoter
rep_origin 5518..5973
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5605..5624)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5815..5836
/label=F1ori-F
/note="F1 origin, forward primer"
polyA_signal 6104..6152
/note="synthetic polyadenylation signal"
misc_feature 6166..6257
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind 6206..6225
/label=RVprimer3
/note="pGL3 vector, forward primer"
ORIGIN
1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatc tgcgatctgc atctcaatta
61 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
121 cgcccattct ccgccccatc gctgactaat tttttttatt tatgcagagg ccgaggccgc
181 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
241 caaaaagctt ggcattccgg tactgttggt aaagccacca tggaagacgc caaaaacata
301 aagaaaggcc cggcgccatt ctatccgctg gaagatggaa ccgctggaga gcaactgcat
361 aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc
421 gaggtggaca tcacttacgc tgagtacttc gaaatgtccg ttcggttggc agaagctatg
481 aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa
541 ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac
601 atttataatg aacgtgaatt gctcaacagt atgggcattt cgcagcctac cgtggtgttc
661 gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa
721 aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc
781 gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtgccaga gtccttcgat
841 agggacaaga caattgcact gatcatgaac tcctctggat ctactggtct gcctaaaggt
901 gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc atgccagaga tcctattttt
961 ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt
1021 ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga
1081 tttgaagaag agctgtttct gaggagcctt caggattaca agattcaaag tgcgctgctg
1141 gtgccaaccc tattctcctt cttcgccaaa agcactctga ttgacaaata cgatttatct
1201 aatttacacg aaattgcttc tggtggcgct cccctctcta aggaagtcgg ggaagcggtt
1261 gccaagaggt tccatctgcc aggtatcagg caaggatatg ggctcactga gactacatca
1321 gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca
1381 ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcaaaga
1441 ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa tccggaagcg
1501 accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac
1561 gaagacgaac acttcttcat cgttgaccgc ctgaagtctc tgattaagta caaaggctat
1621 caggtggctc ccgctgaatt ggaatccatc ttgctccaac accccaacat cttcgacgca
1681 ggtgtcgcag gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg
1741 gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca
1801 accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc
1861 ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag
1921 atcgccgtgg aattcggaag cggagctact aacttcagcc tgctgaagca ggctggagac
1981 gtggaggaga accctggacc tatgacttcg aaagtttatg atccagaaca aaggaaacgg
2041 atgataactg gtccgcagtg gtgggccaga tgtaaacaaa tgaatgttct tgattcattt
2101 attaattatt atgattcaga aaaacatgca gaaaatgctg ttattttttt acatggtaac
2161 gcggcctctt cttatttatg gcgacatgtt gtgccacata ttgagccagt agcgcggtgt
2221 attataccag accttattgg tatgggcaaa tcaggcaaat ctggtaatgg ttcttatagg
2281 ttacttgatc attacaaata tcttactgca tggtttgaac ttcttaattt accaaagaag
2341 atcatttttg tcggccatga ttggggtgct tgtttggcat ttcattatag ctatgagcat
2401 caagataaga tcaaagcaat agttcacgct gaaagtgtag tagatgtgat tgaatcatgg
2461 gatgaatggc ctgatattga agaagatatt gcgttgatca aatctgaaga aggagaaaaa
2521 atggttttgg agaataactt cttcgtggaa accatgttgc catcaaaaat catgagaaag
2581 ttagaaccag aagaatttgc agcatatctt gaaccattca aagagaaagg tgaagttcgt
2641 cgtccaacat tatcatggcc tcgtgaaatc ccgttagtaa aaggtggtaa acctgacgtt
2701 gtacaaattg ttaggaatta taatgcttat ctacgtgcaa gtgatgattt accaaaaatg
2761 tttattgaat cggacccagg attcttttcc aatgctattg ttgaaggtgc caagaagttt
2821 cctaatactg aatttgtcaa agtaaaaggt cttcattttt cgcaagaaga tgcacctgat
2881 gaaatgggaa aatatatcaa atcgttcgtt gagcgagttc tcaaaaatga acaataagaa
2941 ttctagagtc ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt
3001 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
3061 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
3121 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
3181 tacaaatgtg gtaaaatcga taaggatctg aacgatggag cggagaatgg gcggaactgg
3241 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
3301 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg
3361 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
3421 cacaccctaa ctgacacaca ttccacagcg gatccgtcga ccgatgccct tgagagcctt
3481 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
3541 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc
3601 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
3661 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
3721 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
3781 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
3841 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
3901 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
3961 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
4021 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
4081 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
4141 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
4201 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
4261 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
4321 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
4381 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
4441 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
4501 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
4561 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
4621 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
4681 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
4741 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
4801 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
4861 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
4921 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
4981 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
5041 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
5101 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
5161 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
5221 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
5281 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
5341 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
5401 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
5461 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
5521 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
5581 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
5641 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
5701 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
5761 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
5821 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
5881 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
5941 cgaattttaa caaaatatta acgcttacaa tttgccattc gccattcagg ctgcgcaact
6001 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagcccaag ctaccatgat
6061 aagtaagtaa tattaaggta cgggaggtac ttggagcggc cgcaataaaa tatctttatt
6121 ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gtactaacat acgctctcca
6181 tcaaaacaaa acgaaacaaa acaaactagc aaaataggct gtccccagtg caagtgcagg
6241 tgccagaaca tttctctatc gata
//
