质粒名称: | pCAG-Cre-IRES2-GFP |
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出品公司: | 爱迪基因 |
目录编号: | 26646 |
存储实验室: | Anjen Chenn |
载体骨架基本信息 |
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载体名称: | pCAG-IRES2-GFP |
载体抗性: | 氨苄霉素 |
空载体大小: | 6000 bp |
完整质粒大小: | 7000 bp |
载体类型: | 哺乳细胞表达 |
筛选标记: | 新霉素(G418) |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | Cre |
物种来源: | Bacteriophage P1 |
插入大小: | 1000 bp |
标签/融合蛋白: |
IRES2 (C terminal on insert)
GFP (C terminal on insert)
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克隆方法: | 限制性内切酶 |
5' 测序引物 | pCAG-Fwd |
3' 测序引物 | GFP-R |
文献引用方法 |
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材料和方法部分: | pCAG-Cre-IRES2-GFP was a gift from Anjen Chenn (爱迪基因 plasmid # 26646) |
参考文献部分: |
Cell-autonomous beta-catenin signaling regulates cortical precursor proliferation. Woodhead GJ, Mutch CA, Olson EC, Chenn A. J Neurosci. 2006 Nov 29;26(48):12620-30. 10.1523/JNEUROSCI.3180-06.2006 PubMed 17135424 |
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载体基本信息
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载体简介
载体序列
LOCUS pCAG-Cre-IRES2-GFP 7250 bp ds-DNA circular SYN 10-JAN-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCAG-Cre-IRES2-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7250) AUTHORS Woodhead GJ, Mutch CA, Olson EC, Chenn A TITLE Cell-autonomous beta-catenin signaling regulates cortical precursor proliferation. JOURNAL J Neurosci. 2006 Nov 29;26(48):12620-30. PUBMED 17135424 COMMENT This file is created by Vector NTI http://www.biofeng.com/ COMMENT ORIGDB|GenBank COMMENT VNTAUTHORNAME|biofeng.com| COMMENT VNTNAME|pCAG-Cre-IRES2-GFP| FEATURES Location/Qualifiers source 1..7250 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 14..393 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 647..1663 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1587..1609 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1671..1690 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 1798..1818 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1822..2847 /codon_start=1 /product="site-specific recombinase" /label=Cre /note="Cre is a site-specific recombinase from bacteriophage P1. Recombination occurs at loxP sequences." /translation="NLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLLS VCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLPR PSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFLG IAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVERW ISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQRY LAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLEDG D" misc_feature 2888..3474 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(3055..3072) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 3282..3301 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 3475..4194 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3520..3541) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3781..3800) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4128..4149 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(4281..4300) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_site 4346..4401 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal" primer_bind complement(4400..4419) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(4762..4778) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4762..4778) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4775..4797) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4786..4802 /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(4810..4840) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4855..4876 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4935..5130 /label=SV40 promoter /note="SV40 early promoter" rep_origin 4981..5116 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5043..5062 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 5136..5270 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5173..5192) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5227..5246 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5338..5355) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5509..6097) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5589..5608) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6268..7128) /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 6891..6910 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7129..7233) /gene="bla" /label=AmpR promoter ORIGIN 1 tgccacctgg gtcgacattg attattgact agttattaat agtaatcaat tacggggtca 61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct 121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta 181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac 241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt 301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag 361 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc 421 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta 481 ttttgtgcag cgatgggggc gggggggggg ggggggcgag gggcggggcg gggcgaggcg 541 gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag 601 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg 661 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact 721 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta 781 gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct 841 ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt 901 ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc 961 ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg 1021 gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg 1081 agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt 1141 tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc 1201 cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc 1261 cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc 1321 ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt 1381 gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg 1441 cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc 1501 gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg 1561 ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag 1621 agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct 1681 ggttattgtg ctgtctcatc attttggcaa agaattctgc agtcgacggt accgcgggcc 1741 ctcgacctgc agcccaagct gtaccccctc gaaactgaca ggagaaccac catgggccca 1801 aagaagaaga gaaaggtttc gaatttactg accgtacacc aaaatttgcc tgcattaccg 1861 gtcgatgcaa cgagtgatga ggttcgcaag aacctgatgg acatgttcag ggatcgccag 1921 gcgttttctg agcatacctg gaaaatgctt ctgtccgttt gccggtcgtg ggcggcatgg 1981 tgcaagttga ataaccggaa atggtttccc gcagaacctg aagatgttcg cgattatctt 2041 ctatatcttc aggcgcgcgg tctggcagta aaaactatcc agcaacattt gggccagcta 2101 aacatgcttc atcgtcggtc cgggctgcca cgaccaagtg acagcaatgc tgtttcactg 2161 gttatgcggc ggatccgaaa agaaaacgtt gatgccggtg aacgtgcaaa acaggctcta 2221 gcgttcgaac gcactgattt cgaccaggtt cgttcactca tggaaaatag cgatcgctgc 2281 caggatatac gtaatctggc atttctgggg attgcttata acaccctgtt acgtatagcc 2341 gaaattgcca ggatcagggt taaagatatc tcacgtactg acggtgggag aatgttaatc 2401 catattggca gaacgaaaac gctggttagc accgcaggtg tagagaaggc acttagcctg 2461 ggggtaacta aactggtcga gcgatggatt tccgtctctg gtgtagctga tgatccgaat 2521 aactacctgt tttgccgggt cagaaaaaat ggtgttgccg cgccatctgc caccagccag 2581 ctatcaactc gcgccctgga agggattttt gaagcaactc atcgattgat ttacggcgct 2641 aaggatgact ctggtcagag atacctggcc tggtctggac acagtgcccg tgtcggagcc 2701 gcgcgagata tggcccgcgc tggagtttca ataccggaga tcatgcaagc tggtggctgg 2761 accaatgtaa atattgtcat gaactatatc cgtaacctgg atagtgaaac aggggcaatg 2821 gtgcgcctgc tggaagatgg cgattagcca ttaacgcgcg accagcttga tatcgaattg 2881 ggatccgccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag 2941 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga 3001 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg 3061 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt 3121 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca 3181 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc 3241 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat 3301 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc 3361 ctcggtacac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga 3421 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtg 3481 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac 3541 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag 3601 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg 3661 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac 3721 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag 3781 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac 3841 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg 3901 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc 3961 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac 4021 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg 4081 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg 4141 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc 4201 cgcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg 4261 gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag 4321 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt 4381 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat 4441 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac 4501 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta 4561 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt 4621 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc 4681 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag 4741 cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac 4801 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 4861 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 4921 gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc 4981 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt 5041 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga 5101 ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg 5161 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt 5221 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta 5281 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 5341 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 5401 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 5461 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 5521 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 5581 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 5641 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 5701 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 5761 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 5821 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 5881 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 5941 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 6001 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 6061 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 6121 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 6181 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 6241 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 6301 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 6361 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 6421 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 6481 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 6541 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 6601 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 6661 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 6721 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 6781 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 6841 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 6901 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 6961 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 7021 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 7081 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 7141 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 7201 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag //
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