| 质粒名称: | AAV-GFP/Cre |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 49056 |
| 存储实验室: | Fred Gage |
|
载体骨架基本信息 |
|
| 载体名称: | adeno-associated viral (AAVsp) |
| 空载体大小: | 6165 bp |
| 完整质粒大小: | 7956 bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | Cre / GFP-NLS-Cre / green fluorescent protein Cre recombinase (GFP/Cre) fusion protein |
| 物种来源: | M. musculus (mouse); Aequorea victoria green |
| 插入大小: | 1837bp |
| 启动子: | CMV |
| 标签/融合蛋白: |
EGFP (N terminal on insert)
Myc (N terminal on insert)
NLS (N terminal on insert)
|
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | PmlI(未破坏) |
| 3’克隆位点: | XhoI(未破坏) |
| 5’测序引物 | tagccacaccagccaccact |
| 3’测序引物 | caacgtgctggtctgtgtg |
|
文献引用方法 |
|
| 材料和方法部分: | AAV-GFP/Cre was a gift from Fred Gage (Addgene plasmid # 49056 ; http://n2t.net/addgene:49056 ; RRID:Addgene_49056) |
| 参考文献部分: | Adeno-associated virus effectively mediates conditional gene modification in the brain. Kaspar BK, Vissel B, Bengoechea T, Crone S, Randolph-Moore L, Muller R, Brandon EP, Schaffer D, Verma IM, Lee KF, Heinemann SF, Gage FH. Proc Natl Acad Sci U S A. 2002 Feb 19;99(4):2320-5. Epub 2002 Feb 12. 10.1073/pnas.042678699 PubMed 11842206 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS AAV-GFP/Cre 7956 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION adeno-associated virus (AAV) encoding a green fluorescent
protein/Cre recombinase (GFP/Cre) fusion protein..
ACCESSION .
VERSION .
KEYWORDS AAV-GFP/Cre
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7956)
AUTHORS Kaspar BK, Vissel B, Bengoechea T, Crone S, Randolph-Moore L, Muller
R, Brandon EP, Schaffer D, Verma IM, Lee KF, Heinemann SF, Gage FH
TITLE Adeno-associated virus effectively mediates conditional gene
modification in the brain.
JOURNAL Proc Natl Acad Sci U S A. 2002 Feb 19;99(4):2320-5. Epub 2002 Feb
12.
PUBMED 11842206
REFERENCE 2 (bases 1 to 7956)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7956
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 7..136
/label=AAV2 ITR
primer_bind complement(198..217)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
enhancer 319..698
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 699..902
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 853..873
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 899..923
/label=LNCX
/note="Human CMV promoter, forward primer"
intron 1036..1511
/label=-beta-globin intron
/note="internally truncated intron from human beta-globin"
regulatory 1585..1594
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1591..2307
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 1591..2307
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1636..1657)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1897..1916)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2244..2265
/label=EGFP-C
/note="EGFP, forward primer"
CDS 2314..2343
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 2350..2370
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2371..3399
/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="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
GD"
polyA_signal 3487..3881
/label=-beta-globin poly(A)
/note="human beta-globin polyadenylation signal"
repeat_region 4242..4371
/label=AAV2 ITR
rep_origin 4742..5255
/direction=RIGHT
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4829..4848)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5039..5060
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(5737..5756)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5856..5878
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5916..5934)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6002..6106
/gene="bla"
/label=AmpR promoter
CDS 6107..6967
/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(6325..6344)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7138..7726
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7627..7646
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7880..7897
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 atgcagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
61 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
121 atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca
181 tgctctagag gatccggcct cggcctctgc ataaataaaa aaaattagtc agccatgagc
241 ttggcccatt gcatacgttg tatccatatc ataatatgta catttatatt ggctcatgtc
301 caacattacc gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
361 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
421 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
481 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
541 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
601 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
661 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
721 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
781 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
841 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
901 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
961 gaagacaccg ggaccgatcc agcctcccct cgaagcttac atgtggtacc gagctcggat
1021 cctgagaact tcagggtgag tctatgggac ccttgatgtt ttctttcccc ttcttttcta
1081 tggttaagtt catgtcatag gaaggggaga agtaacaggg tacacatatt gaccaaatca
1141 gggtaatttt gcatttgtaa ttttaaaaaa tgctttcttc ttttaatata cttttttgtt
1201 tatcttattt ctaatacttt ccctaatctc tttctttcag ggcaataatg atacaatgta
1261 tcatgcctct ttgcaccatt ctaaagaata acagtgataa tttctgggtt aaggcaatag
1321 caatatttct gcatataaat atttctgcat ataaattgta actgatgtaa gaggtttcat
1381 attgctaata gcagctacaa tccagctacc attctgcttt tattttatgg ttgggataag
1441 gctggattat tctgagtcca agctaggccc ttttgctaat catgttcata cctcttatct
1501 tcctcccaca gctcctgggc aacgtgctgg tctgtgtgct ggcccatcac tttggcaaag
1561 cacgtgaatt cgatgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
1621 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
1681 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
1741 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
1801 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
1861 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
1921 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
1981 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
2041 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
2101 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
2161 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
2221 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
2281 actctcggca tggacgagct gtacaagtcc ggagagcaaa agctgatttc tgaggaggat
2341 ctgggaggac ccaagaagaa gaggaaggtg tccaatttac tgaccgtaca ccaaaatttg
2401 cctgcattac cggtcgatgc aacgagtgat gaggttcgca agaacctgat ggacatgttc
2461 agggatcgcc aggcgttttc tgagcatacc tggaaaatgc ttctgtccgt ttgccggtcg
2521 tgggcggcat ggtgcaagtt gaataaccgg aaatggtttc ccgcagaacc tgaagatgtt
2581 cgcgattatc ttctatatct tcaggcgcgc ggtctggcag taaaaactat ccagcaacat
2641 ttgggccagc taaacatgct tcatcgtcgg tccgggctgc cacgaccaag tgacagcaat
2701 gctgtttcac tggttatgcg gcggatccga aaagaaaacg ttgatgccgg tgaacgtgca
2761 aaacaggctc tagcgttcga acgcactgat ttcgaccagg ttcgttcact catggaaaat
2821 agcgatcgct gccaggatat acgtaatctg gcatttctgg ggattgctta taacaccctg
2881 ttacgtatag ccgaaattgc caggatcagg gttaaagata tctcacgtac tgacggtggg
2941 agaatgttaa tccatattgg cagaacgaaa acgctggtta gcaccgcagg tgtagagaag
3001 gcacttagcc tgggggtaac taaactggtc gagcgatgga tttccgtctc tggtgtagct
3061 gatgatccga ataactacct gttttgccgg gtcagaaaaa atggtgttgc cgcgccatct
3121 gccaccagcc agctatcaac tcgcgccctg gaagggattt ttgaagcaac tcatcgattg
3181 atttacggcg ctaaggatga ctctggtcag agatacctgg cctggtctgg acacagtgcc
3241 cgtgtcggag ccgcgcgaga tatggcccgc gctggagttt caataccgga gatcatgcaa
3301 gctggtggct ggaccaatgt aaatattgtc atgaactata tccgtaacct ggatagtgaa
3361 acaggggcaa tggtgcgcct gctggaagat ggcgattagc tcgagcaatt caccccacca
3421 gtgcaggctg cctatcagaa agtggtggct ggtgtggcta atgccctggc ccacaagtat
3481 cactaagctc gctttcttgc tgtccaattt ctattaaagg ttcctttgtt ccctaagtcc
3541 aactactaaa ctgggggata ttatgaaggg ccttgagcat ctggattctg cctaataaaa
3601 aacatttatt ttcattgcaa tgatgtattt aaattatttc tgaatatttt actaaaaagg
3661 gaatgtggga ggtcagtgca tttaaaacat aaagaaatga agagctagtt caaaccttgg
3721 gaaaatacac tatatcttaa actccatgaa agaaggtgag gctgcaaaca gctaatgcac
3781 attggcaaca gcccctgatg cctatgcctt attcatccct cagaaaagga ttcaagtaga
3841 ggcttgattt ggaggttaaa gttttgctat gctgtatttt acattactta ttgttttagc
3901 tgtcctcatg aatgtctttt cactacccat ttgcttatcc tgcatctctc agccttgact
3961 ccactcagtt ctcttgctta gagataccac ctttcccctg aagtgttcct tccatgtttt
4021 acggcgagat ggtttctcct cgcctggcca ctcagcctta gttgtctctg ttgtcttata
4081 gaggtctact tgaagaagga aaaacagggg gcatggtttg actgtcctgt gagcccttct
4141 tccctgcctc ccccactcac agtgacccgg aatccctcga catggcagtc tagagcatgg
4201 ctacgtagat aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga
4261 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
4321 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctggcgtaa
4381 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
4441 gcgattccgt tgcaatggct ggcggtaata ttgttctgga tattaccagc aaggccgata
4501 gtttgagttc ttctactcag gcaagtgatg ttattactaa tcaaagaagt attgcgacaa
4561 cggttaattt gcgtgatgga cagactcttt tactcggtgg cctcactgat tataaaaaca
4621 cttctcagga ttctggcgta ccgttcctgt ctaaaatccc tttaatcggc ctcctgttta
4681 gctcccgctc tgattctaac gaggaaagca cgttatacgt gctcgtcaaa gcaaccatag
4741 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
4801 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
4861 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
4921 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
4981 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
5041 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
5101 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
5161 aacgcgaatt ttaacaaaat attaacgctt acaatttaaa tatttgctta tacaatcttc
5221 ctgtttttgg ggcttttctg attatcaacc ggggtacata tgattgacat gctagtttta
5281 cgattaccgt tcatcgattc tcttgtttgc tccagactct caggcaatga cctgatagcc
5341 tttgtagaga cctctcaaaa atagctaccc tctccggcat gaatttatca gctagaacgg
5401 ttgaatatca tattgatggt gatttgactg tctccggcct ttctcacccg tttgaatctt
5461 tacctacaca ttactcaggc attgcattta aaatatatga gggttctaaa aatttttatc
5521 cttgcgttga aataaaggct tctcccgcaa aagtattaca gggtcataat gtttttggta
5581 caaccgattt agctttatgc tctgaggctt tattgcttaa ttttgctaat tctttgcctt
5641 gcctgtatga tttattggat gttggaatcg cctgatgcgg tattttctcc ttacgcatct
5701 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
5761 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
5821 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
5881 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
5941 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
6001 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
6061 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
6121 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
6181 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
6241 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
6301 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
6361 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
6421 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
6481 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
6541 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
6601 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
6661 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
6721 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
6781 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
6841 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
6901 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
6961 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
7021 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
7081 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
7141 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
7201 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
7261 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
7321 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
7381 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
7441 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
7501 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
7561 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
7621 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
7681 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
7741 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
7801 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
7861 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
7921 caaaccgcct ctccccgcgc gttggccgat tcatta
//
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