| 质粒名称: | CRE recombinase |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 62730 |
| 存储实验室: | Niels Geijsen |
|
载体骨架基本信息 |
|
| 载体名称: | pET-15 |
| 空载体大小: | 5646 bp |
| 完整质粒大小: | 6786 bp |
| 载体类型: | 细菌载体表达, Bacterial Expression, Cre/Lox |
| 载体抗性: | Ampicillin, 100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Cre recombinase |
| 物种来源: | P1 Bacteriophage |
| 插入大小: | 1134bp |
| 基因突变: | Codon optimazed for e.coli |
| 启动子: | T7 |
| 标签/融合蛋白: |
His tag (N terminal on insert)
NLS (N terminal on insert)
|
| 克隆方法: | Ligation Independent Cloning |
| 5’测序引物 | GATCGAGATCTCGATCCCGCG |
| 3’测序引物 | TTTCAGCAAAAAACCCCTCAAGACC |
|
文献引用方法 |
|
| 材料和方法部分: | CRE recombinase was a gift from Niels Geijsen (Addgene plasmid # 62730 ; http://n2t.net/addgene:62730 ; RRID:Addgene_62730) |
| 参考文献部分: | Efficient Intracellular Delivery of Native Proteins. D'Astolfo DS, Pagliero RJ, Pras A, Karthaus WR, Clevers H, Prasad V, Lebbink RJ, Rehmann H, Geijsen N. Cell. 2015 Apr 23;161(3):674-690. doi: 10.1016/j.cell.2015.03.028. 10.1016/j.cell.2015.03.028 PubMed 25910214 |
买家导航
CRE recombinase载体质粒基本信息
CRE recombinase质粒图谱载体图谱和CRE recombinase载体序列质粒序列多克隆位点信息
CRE recombinase质粒载体简介
CRE recombinase质粒序列载体序列
LOCUS CRE recombinase 6783 bp ds-DNA circular SYN 18-JAN-2022
DEFINITION Expresses Cre recombinase protein in bacterial cells.
ACCESSION .
VERSION .
KEYWORDS CRE recombinase
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6783)
AUTHORS D'Astolfo DS, Pagliero RJ, Pras A, Karthaus WR, Clevers H, Prasad V,
Lebbink RJ, Rehmann H, Geijsen N
TITLE Efficient Intracellular Delivery of Native Proteins.
JOURNAL Cell. 2015 Apr 23;161(3):674-690. doi: 10.1016/j.cell.2015.03.028.
PUBMED 25910214
REFERENCE 2 (bases 1 to 6783)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jan 18, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6783
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(45..63)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 131..235
/gene="bla"
/label=AmpR promoter
CDS 236..1096
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(454..473)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1267..1855
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1756..1775
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2009..2026
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 2041..2180
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(2106..2125)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2266..2288
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(2282..2473)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(3785..4867)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 4829..4848
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(4868..4945)
/gene="lacI"
/label=lacI promoter
primer_bind 5151..5170
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 5254..5273
/label=T7
/note="T7 promoter, forward primer"
promoter 5254..5272
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 5273..5297
/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)."
RBS 5312..5334
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 5353..5370
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 5380..5397
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS 5437..5457
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(6537..6555)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 6551..6598
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter complement(6773..18)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
ORIGIN
1 tcgatgataa gctgtcaaac atgagaattc ttgaagacga aagggcctcg tgatacgcct
61 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
121 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
181 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
241 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
301 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
361 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
421 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
481 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
541 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
601 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
661 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
721 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
781 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
841 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
901 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
961 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
1021 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
1081 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
1141 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
1201 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
1261 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
1321 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
1381 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
1441 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
1501 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
1561 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
1621 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
1681 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
1741 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
1801 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
1861 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
1921 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
1981 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
2041 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
2101 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
2161 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
2221 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
2281 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
2341 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
2401 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
2461 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
2521 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
2581 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
2641 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
2701 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
2761 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
2821 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
2881 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
2941 gccaggaccc aacgctgccc gagatgcgcc gcgtgcggct gctggagatg gcggacgcga
3001 tggatatgtt ctgccaaggg ttggtttgcg cattcacagt tctccgcaag aattgattgg
3061 ctccaattct tggagtggtg aatccgttag cgaggtgccg ccggcttcca ttcaggtcga
3121 ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt atagggcggc
3181 gcctacaatc catgccaacc cgttccatgt gctcgccgag gcggcataaa tcgccgtgac
3241 gatcagcggt ccaatgatcg aagttaggct ggtaagagcc gcgagcgatc cttgaagctg
3301 tccctgatgg tcgtcatcta cctgcctgga cagcatggcc tgcaacgcgg gcatcccgat
3361 gccgccggaa gcgagaagaa tcataatggg gaaggccatc cagcctcgcg tcgcgaacgc
3421 cagcaagacg tagcccagcg cgtcggccgc catgccggcg ataatggcct gcttctcgcc
3481 gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa
3541 taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat
3601 gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag
3661 tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct
3721 caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt
3781 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
3841 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca
3901 gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc
3961 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga
4021 tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc
4081 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca
4141 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca
4201 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt
4261 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg
4321 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat
4381 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa
4441 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga
4501 tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc
4561 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa
4621 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca
4681 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg
4741 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca
4801 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg
4861 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa
4921 aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc
4981 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
5041 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
5101 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
5161 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
5221 gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt
5281 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc
5341 atgggctcat cgcatcatca tcatcatcat tcatcgggtc tggttccgcg cggttcacac
5401 ggtggtggtt cggcggccgc tatgggcacc cgcctgccga aaaaaaaacg caaagtgtcc
5461 aacctgctga cggtgcacca gaatctgccg gcactgccgg tcgatgcaac cagtgacgaa
5521 gtgcgcaaaa atctgatgga tatgtttcgt gaccgccaag ccttcagcga acatacgtgg
5581 aaaatgctgc tgtcggtttg ccgtagctgg gcggcctggt gtaaactgaa caatcgcaaa
5641 tggtttccgg cagaaccgga agatgtgcgt gactatctgc tgtacctgca ggcacgtggt
5701 ctggcagtta aaaccatcca gcaacatctg ggccaactga acatgctgca ccgtcgctct
5761 ggtctgccgc gtccgagtga ttccaatgcc gtcagtctgg tgatgcgtcg cattcgtaaa
5821 gaaaacgtgg atgcaggcga acgcgctaaa caggcactgg cttttgaacg taccgatttc
5881 gaccaagttc gctctctgat ggaaaacagt gatcgttgcc aggacatccg caatctggca
5941 ttcctgggta ttgcttataa caccctgctg cgcattgcag aaatcgctcg tattcgcgtg
6001 aaagatatca gccgtacgga cggcggtcgc atgctgattc acatcggccg taccaaaacg
6061 ctggtttcca ccgcgggcgt cgaaaaagcc ctgtcactgg gtgtcacgaa actggtggaa
6121 cgctggattt cagtttcggg cgtcgcagat gacccgaaca attacctgtt ttgtcgtgtg
6181 cgcaaaaatg gtgttgcagc tccgagcgct acctctcagc tgagtacgcg tgcgctggaa
6241 ggcatcttcg aagccaccca tcgcctgatt tatggcgcga aagatgacag cggtcagcgt
6301 tacctggcat ggtccggtca ctcagctcgt gttggtgcag cacgtgatat ggcacgtgca
6361 ggtgtctcta tcccggaaat tatgcaggcc ggcggttgga cgaacgtgaa tattgttatg
6421 aactatattc gtaacctgga ctctgaaacg ggtgcgatgg tgcgtctgct ggaagatggc
6481 gactgaggat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
6541 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
6601 gaaaggagga actatatccg gatatcccgc aagaggcccg gcagtaccgg cataaccaag
6661 cctatgccta cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat
6721 ttcatacacg gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc
6781 tta
//
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