| 质粒名称: | GST-BHMT-I/G |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 104442 |
| 存储实验室: | Carol Mercer |
|
载体骨架基本信息 |
|
| 载体名称: | pRK5 |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4754 bp |
| 完整质粒大小: | 7832 bp |
| 载体类型: | 哺乳细胞表达 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | BHMT |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 3213 bp |
| 基因突变: | GST中的N43A（潜在的KFERQ样序列）。BHMT不包含前8个或后9个氨基酸。 |
| 启动子 | CMV |
| 标签/融合蛋白: |
GST（插入件上的N端子）
GFP myc GFP来自Aeqourea victoria（插接件上的C端子）
|
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Nru1（销毁后未知） |
| 3' 克隆位点: | HindIII（销毁后未知） |
| 5' 测序引物 | SP6 5'-TGATGAGTTTGGACAAACCAC-3' |
|
文献引用方法 |
|
| 材料和方法部分: | GST-BHMT-I/G was a gift from Carol Mercer (Addgene plasmid # 104442 ; http://n2t.net/addgene:104442 ; RRID:Addgene_104442) |
| 参考文献部分: | Macroautophagy-dependent, intralysosomal cleavage of a betaine homocysteine methyltransferase fusion protein requires stable multimerization. Mercer CA, Kaliappan A, Dennis PB. Autophagy. 2008 Feb;4(2):185-94. Epub 2007 Nov 7. 5275 [pii] PubMed 18059170 |
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 GST-BHMT-I/G 7832 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Autophagy cargo assay.
ACCESSION .
VERSION .
KEYWORDS GST-BHMT-I/G
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7832)
AUTHORS Mercer CA, Kaliappan A, Dennis PB
TITLE Macroautophagy-dependent, intralysosomal cleavage of a betaine
homocysteine methyltransferase fusion protein requires stable
multimerization.
JOURNAL Autophagy. 2008 Feb;4(2):185-94. Epub 2007 Nov 7.
PUBMED 18059170
REFERENCE 2 (bases 1 to 7832)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7832
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 181..201
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
polyA_signal 293..427
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(317..336)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 371..390
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
regulatory 431..440
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS complement(451..480)
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS complement(490..1206)
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(532..553)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 881..900
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1140..1161
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1368..1387)
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
primer_bind 1597..1614
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind complement(3013..3035)
/label=pGEX 5'
/note="pGEX vectors, Glutathione-S-transferase, forward
primer"
primer_bind complement(3694..3713)
/label=pMT2-F
/note="Synthetic intron, forward primer"
promoter complement(3726..3744)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(3727..3744)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(3954..3978)
/label=LNCX
/note="Human CMV promoter, forward primer"
promoter complement(3975..4178)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(4004..4024)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 4179..4558
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind complement(4587..4603)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4587..4603)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4600..4622)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4611..4627
/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(4635..4665)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4680..4701
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4818..4835)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4989..5577)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5069..5088)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5748..6608)
/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 6371..6390
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6609..6713)
/gene="bla"
/label=AmpR promoter
primer_bind 6781..6799
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(6837..6859)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 6959..6978
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
rep_origin complement(6995..7450)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7132..7153)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 7344..7363
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7648..7670
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 7662..7679
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 7663..7679
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(7699..224)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 7713..16
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(7767..7786)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
ORIGIN
1 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
61 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
121 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
181 accctaactg acacacattc cacagctggt tctttccgcc tcagaaggta cctaaccaag
241 ttcctctttc agaggttatt tcaggccatg gtgctgcgcc gaattaattc ccgatccaga
301 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
361 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
421 acaagttggg ccatggcggc caagctttca caggtcctcc tcggagatca gcttctgctc
481 cattctagac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc
541 cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg actgggtgct
601 caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta
661 gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt
721 gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc
781 cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg
841 gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc
901 cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc
961 gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt
1021 cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag
1081 cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac
1141 gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct
1201 caccatggta ttatcgtgtt tttcaaagga aaaccacgtc cccgtggttc ggggggccta
1261 gacgtttttt taacctcgac taaacacatg taaagcatgt gcaccgaggc cccagatcag
1321 atcccataca atggggtacc ttctgggcat ccttcagccc cttgttgaat acgcttgagg
1381 agagccattt gactctttcc acaactatcc aactcacaac gtggcactgg ggttgtgccg
1441 cctttgcagg tgtatcttat acacgtggct tttggccgca gaggcacctg tcgccaggtg
1501 gggggttccg ctgcctgcaa agggtcgcta cagacgttgt ttgtcttcaa gaagcttcca
1561 gaggaactgc ttccttcacg acattcaaca gaccttgcat tcctttggcg agaggggaaa
1621 gacccctagg aatgctcgtc aagaagacag ggccaggttt ccgggccctc acattgccaa
1681 aagacggcaa tatggtggaa aataacatat agacaaacgc acaccggcct tattccaagc
1741 ggcttcggcc agtaacgtta gggggggggc ggaattgatc ccgctcgagg ttaacgaatt
1801 cttatctaga gagctctttc agctgctgct cagttgtggc ttctttctgc tgcatcagct
1861 cggctgttcc tttggtcact ccccagccat ctggctttga cattgaaggg ttgtatggcc
1921 ggcctgaggc tatccgaaga ttctcccagt attccttcct ggcccttgct ctaacccagg
1981 gtttggtgtg catgtccaaa ccacttcccc agctgccatg tttttctgaa gctggtggca
2041 aaaagcccct ttctggggcc agctcctctg caattgccct gatgtggtag ggctcaaatc
2101 cacagcaccc gccaatgtac ctgaccccca ggttgtaggc ctctctggcg tatttttgaa
2161 tatcccatct ggtggcaact ctgggttcca gtccaaatgg gaattctggg agatcgatga
2221 atccctgctt gttgcagtca ggagtgtggt aagccaaggg ctggctcatc aggtgagctt
2281 tcagtcgggc agcctccaag ccctccttca tgagcttcac tgtttttaaa ctaatggtgg
2341 ggtcaaagtg gcagttcaca ccaatgatgg atgctcctgc tttcaccagg cgcactgcac
2401 actcgccggg gggcacgcca tgcaaatctc cttctgggcc aatgcacatg gttgctgcca
2461 caggtttacc ggatgctatc aaggtttcaa ctgcccacac agcttcttca acgtgttcaa
2521 aatactctgc aatcaagaag tccacgttct tcttcataaa gacctctaac tgttgcagaa
2581 atactttttt gacttcagtt tcactcttgc agctaaggta tgaaggtgtc tgactcactc
2641 ctcctgctac caaagcatct ccttcatcag ccacttgtcg ggcgatgtcg caagcagctt
2701 cattgacttc ctgcccagat atcttctcta agacatagtt gcccctgttc tccagcttgt
2761 cttcactcgc atagaaggtg aaggtctgca tgacgtttga gccagctctg aggaactctc
2821 gatgaagctg gcgaactgct tctgggtgct ccacagcagc ttcaggagtc cagggtcctg
2881 cctttacgta gcccctcttc tccagtgcaa agacaaaccc tccatctcca atcacaatct
2941 ctccagcatt taaacgttct aggatgccct tcttggcgcg gccgccaccg gattgaggtg
3001 ggtggccgcc accaccaaac gtggcttgcc agccctgcaa aggccatgct atatacttgc
3061 tggatttcaa gtacttatca atttgtggga tagcttcaat acgtttttta aaacaaacta
3121 attttgggaa cgcatccagg cacattgggt ccatgtataa aacaacatca agagcgtcat
3181 acaacatgaa gtcaggatgg gttacatgat caccatttaa atatgtttta tgacataaac
3241 gatcttcgaa cattttcagc atttcaggta gcttgctaag aaaatcaact ttgagagttt
3301 caaagtcttt actatatgca attctcgaaa caccgtatct aatatccaaa accgctcctt
3361 caagcattga aatctctgca cgctcttttg gacaaccacc caacatgttg tgcttgtcag
3421 ctatataacg tatgatggcc atagactgtg ttaatttaac atcaccatca atataataag
3481 gaagattggg aaactccaaa cccaattcaa actttttggc tcgccattta tcaccttcat
3541 cgcgctcata caaatgctct tcatattttt cttcaagata ttccaaaaga agtcgagtgg
3601 gttgcacaag gcccttaatt ttccaataac ctagtatagg ggacatggct ggctcgcgat
3661 agaaccgagg tgcagttgga cctgggagtg gacacctgtg gagagaaagg caaagtggat
3721 gttattctat agtgtcacct aaatcgtatg tgtatgatac ataaggttat gtattaattg
3781 tagccgcgtt ctaacgaagc caagggggtg ggcctataga ctctataggc ggtacttacg
3841 tcactcttgg cacggggaat ccgcgttcca atgcaccgtt cccggccgcg gaggctggat
3901 cggtcccggt gtcttctatg gaggtcaaaa cagcgtggat ggcgtctcca ggcgatctga
3961 cggttcacta aacgagctct gcttatatag acctcccacc gtacacgcct accgcccatt
4021 tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa
4081 aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc
4141 tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat
4201 acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag
4261 gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga
4321 tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc
4381 cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg
4441 ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg
4501 ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcgg
4561 gcgagctcga attaattcgt aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg
4621 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
4681 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
4741 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
4801 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
4861 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
4921 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
4981 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
5041 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
5101 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
5161 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
5221 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
5281 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
5341 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
5401 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
5461 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
5521 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
5581 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
5641 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
5701 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
5761 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
5821 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
5881 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
5941 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
6001 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
6061 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
6121 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
6181 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
6241 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
6301 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
6361 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
6421 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
6481 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
6541 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
6601 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
6661 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
6721 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
6781 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
6841 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
6901 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
6961 gcatcagagc agattgtact gagagtgcac cataaaattg taaacgttaa tattttgtta
7021 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
7081 aaaatccctt ataaatcaaa agaatagccc gagatagggt tgagtgttgt tccagtttgg
7141 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
7201 cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc
7261 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
7321 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
7381 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
7441 cagggcgcgt actatggttg ctttgacgta tgcggtgtga aataccgcac agatgcgtaa
7501 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
7561 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
7621 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
7681 ccaagctgtt aacagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt
7741 ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa aaattagtca
7801 gccatggggc ggagaatggg cggaactggg cg
//
