| 质粒名称: | pCS-kI |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 51553 |
| 存储实验室: | Michele Calos |
|
载体骨架基本信息 |
|
| 载体名称: | pCMVSPORTbetaGal |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | phiC31 integrase |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pCS-kI was a gift from Michele Calos (Addgene plasmid # 51553 ; http://n2t.net/addgene:51553 ; RRID:Addgene_51553) |
| 参考文献部分: | DICE, an efficient system for iterative genomic editing in human pluripotent stem cells. Zhu F, Gamboa M, Farruggio AP, Hippenmeyer S, Tasic B, Schule B, Chen-Tsai Y, Calos MP. Nucleic Acids Res. 2013 Dec 4. 10.1093/nar/gkt1290 PubMed 24304893 |
买家导航
pCSkI载体基本信息
pCSkI载体图谱质粒图谱和多克隆位点信息
pCSkI载体简介
pCSkI载体序列
LOCUS pCS-kI 6204 bp ds-DNA circular SYN 28-OCT-2019
DEFINITION Mammalian phiC31 integrase expression vector. For use in
applications where pseudosite integration is not desired..
ACCESSION .
VERSION .
KEYWORDS pCS-kI
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6204)
AUTHORS Zhu F, Gamboa M, Farruggio AP, Hippenmeyer S, Tasic B, Schule B,
Chen-Tsai Y, Calos MP
TITLE DICE, an efficient system for iterative genomic editing in human
pluripotent stem cells.
JOURNAL Nucleic Acids Res. 2013 Dec 4.
PUBMED 24304893
REFERENCE 2 (bases 1 to 6204)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Oct 28, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6204
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 2..456
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(89..108)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 298..319
/label=F1ori-F
/note="F1 origin, forward primer"
CDS 743..1603
/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(961..980)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
protein_bind complement(1705..1738)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
rep_origin 1816..2404
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2305..2324
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2558..2575
/label=L4440
/note="L4440 vector, forward primer"
primer_bind 2821..2839
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 2866..2896
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
protein_bind 2904..2920
/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 2909..2931
/label=M13/pUC Reverse
/note="In lacZ gene"
enhancer 2950..3253
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3254..3457
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 3408..3428
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 3454..3478
/label=LNCX
/note="Human CMV promoter, forward primer"
promoter 3573..3591
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 3573..3590
/label=SP6
/note="SP6 promoter, forward primer"
regulatory 3654..3663
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3660..5477
/codon_start=1
/product="integrase from phage phi-C31"
/label=phage phi-C31 integrase
/translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
EKRASITWAKPPTDDDEDDAQDGTEDVAA"
primer_bind complement(5540..5559)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(5541..5559)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 5946..6080
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5983..6002)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6037..6056
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
ORIGIN
1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg
301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
421 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
481 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgcc
541 aggtcttgga ctggtgagaa cggcttgctc ggcagcttcg atgtgtgctg gagggagaat
601 aaaggtctaa gatgtgcgat agagggaagt cgcattgaat tatgtgctgt gtagggatcg
661 ctggtatcaa atatgtgtgc ccacccctgg catgagacaa taaccctgat aaatgcttca
721 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
781 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
841 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
901 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
961 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
1021 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
1081 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
1141 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
1201 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
1261 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
1321 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
1381 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
1441 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
1501 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
1561 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
1621 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
1681 gatccttttt gataatctca tgccataact tcgtataatg tatgctatac gaagttatgg
1741 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
1801 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
1861 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
1921 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
1981 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
2041 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
2101 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
2161 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
2221 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
2281 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
2341 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
2401 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
2461 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
2521 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
2581 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
2641 agcttgcaat tcgcgctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
2701 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
2761 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
2821 aataggcgta tcacgaggcc ctttcactca ttaggcaccc caggctttac actttatgct
2881 tccggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcat
2941 cgtgcaggtc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
3001 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
3061 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
3121 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
3181 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
3241 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
3301 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
3361 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
3421 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc
3481 ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct
3541 ccggactcta gcctaggctt ttgcaaaaag ctatttaggt gacactatag aaggtacgcc
3601 tgcaggtacc ggtccggaat tcccgggtcg acgagctcac tagtcgtagg gtcgccacca
3661 tggacacgta cgcgggtgct tacgaccgtc agtcgcgcga gcgcgaaaat tcgagcgcag
3721 caagcccagc gacacagcgt agcgccaacg aagacaaggc ggccgacctt cagcgcgaag
3781 tcgagcgcga cgggggccgg ttcaggttcg tcgggcattt cagcgaagcg ccgggcacgt
3841 cggcgttcgg gacggcggag cgcccggagt tcgaacgcat cctgaacgaa tgccgcgccg
3901 ggcggctcaa catgatcatt gtctatgacg tgtcgcgctt ctcgcgcctg aaggtcatgg
3961 acgcgattcc gattgtctcg gaattgctcg ccctgggcgt gacgattgtt tccactcagg
4021 aaggcgtctt ccggcaggga aacgtcatgg acctgattca cctgattatg cggctcgacg
4081 cgtcgcacaa agaatcttcg ctgaagtcgg cgaagattct cgacacgaag aaccttcagc
4141 gcgaattggg cgggtacgtc ggcgggaagg cgccttacgg cttcgagctt gtttcggaga
4201 cgaaggagat cacgcgcaac ggccgaatgg tcaatgtcgt catcaacaag cttgcgcact
4261 cgaccactcc ccttaccgga cccttcgagt tcgagcccga cgtaatccgg tggtggtggc
4321 gtgagatcaa gacgcacaaa caccttccct tcaagccggg cagtcaagcc gccattcacc
4381 cgggcagcat cacggggctt tgtaagcgca tggacgctga cgccgtgccg acccggggcg
4441 agacgattgg gaagaagacc gcttcaagcg cctgggaccc ggcaaccgtt atgcgaatcc
4501 ttcgggaccc gcgtattgcg ggcttcgccg ctgaggtgat ctacaagaag aagccggacg
4561 gcacgccgac cacgaagatt gagggttacc gcattcagcg cgacccgatc acgctccggc
4621 cggtcgagct tgattgcgga ccgatcatcg agcccgctga gtggtatgag cttcaggcgt
4681 ggttggacgg cagggggcgc ggcaaggggc tttcccgggg gcaagccatt ctgtccgcca
4741 tggacaagct gtactgcgag tgtggcgccg tcatgacttc gaagcgcggg gaagaatcga
4801 tcaaggactc ttaccgctgc cgtcgccgga aggtggtcga cccgtccgca cctgggcagc
4861 acgaaggcac gtgcaacgtc agcatggcgg cactcgacaa gttcgttgcg gaacgcatct
4921 tcaacaagat caggcacgcc gaaggcgacg aagagacgtt ggcgcttctg tgggaagccg
4981 cccgacgctt cggcaagctc actgaggcgc ctgagaagag cggcgaacgg gcgaaccttg
5041 ttgcggagcg cgccgacgcc ctgaacgccc ttgaagagct gtacgaagac cgcgcggcag
5101 gcgcgtacga cggacccgtt ggcaggaagc acttccggaa gcaacaggca gcgctgacgc
5161 tccggcagca aggggcggaa gagcggcttg ccgaacttga agccgccgaa gccccgaagc
5221 ttccccttga ccaatggttc cccgaagacg ccgacgctga cccgaccggc cctaagtcgt
5281 ggtgggggcg cgcgtcagta gacgacaagc gcgtgttcgt cgggctcttc gtagacaaga
5341 tcgttgtcac gaagtcgact acgggcaggg ggcagggaac gcccatcgag aagcgcgctt
5401 cgatcacgtg ggcgaagccg ccgaccgacg acgacgaaga cgacgcccag gacggcacgg
5461 aagacgtagc ggcgtagcga gacacccgga tccctcgagg ggcccaagct tacgcgtgca
5521 tgcgacgtca tagctctctc cctatagtga gtcgtattat aagctagctt gggatctttg
5581 tgaaggaacc ttacttctgt ggtgtgacat aattggacaa actacctaca gagatttaaa
5641 gctctaaggt aaatataaaa tttttaagtg tataatgtgt taaactagct gcatatgctt
5701 gctgcttgag agttttgctt actgagtatg atttatgaaa atattataca caggagctag
5761 tgattctaat tgtttgtgta ttttagattc acagtcccaa ggctcatttc aggcccctca
5821 gtcctcacag tctgttcatg atcataatca gccataccac atttgtagag gttttacttg
5881 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
5941 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
6001 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
6061 tatcttatca tgtctggatc gatcctgcat taatgaatcg gccaacgcgc ggggagaggc
6121 ggtttgcgta ttggctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
6181 ttgcgcagcc tgaatggcga atgg
//
