| 质粒名称: | pET His6 Sumo TEV LIC cloning vector (2S-T) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 29711 |
| 存储实验室: | Scott Gradia |
|
载体骨架基本信息 |
|
| 载体名称: | pET |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5104 bp |
| 载体类型: | 细菌表达 |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 标签/融合蛋白: | His6 Sumo TEV（主干上的N终端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | LIC site（在克隆过程中被破坏） |
| 3' 克隆位点: | LIC site（在克隆过程中被破坏） |
| 5' 测序引物 | Sumo forward (5'ttattgaggctcacagagaac) |
| 3' 测序引物 | T7 reverse |
|
文献引用方法 |
|
| 材料和方法部分: | pET His6 Sumo TEV LIC cloning vector (2S-T) was a gift from Scott Gradia (Addgene plasmid # 29711 ; http://n2t.net/addgene:29711 ; RRID:Addgene_29711) |
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
质粒图谱.png)
载体简介
载体序列
LOCUS pET His6 Sumo TEV LIC cloning vector (2S-T) 5101 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pET His6 Sumo TEV LIC cloning vector (2S-T)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5101)
TITLE 2-series LIC N-terminal fusion vectors for E. coli expression
REFERENCE 2 (bases 1 to 5101)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5101
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 1..20
/label=T7
/note="T7 promoter, forward primer"
promoter 1..19
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 108..130
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 150..167
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 186..476
/codon_start=1
/gene="S. cerevisiae SMT3 (truncated)"
/product="cleavable ubiquitin-like protein tag"
/label=SUMO
/translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIG"
CDS 486..506
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
primer_bind complement(631..649)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 645..692
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter complement(1058..1086)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
primer_bind complement(1113..1131)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1199..1303
/gene="bla"
/label=AmpR promoter
CDS 1304..2164
/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(1522..1541)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2335..2923
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2824..2843
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3077..3094
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 3109..3248
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(3174..3193)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3334..3356
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(3350..3541)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind 4999..5018
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
ORIGIN
1 taatacgact cactataggg agaccacaac ggtttccctc tagtgccggc tccggagagc
61 tctttaatta agcggccgcc ctgcaggact cgagttctag aaataatttt gtttaacttt
121 aagaaggaga tatacatatg aaatcttctc accatcacca tcaccatggt tcttctatgg
181 ctagcatgtc ggactcagaa gtcaatcaag aagctaagcc agaggtcaag ccagaagtca
241 agcctgagac tcacatcaat ttaaaggtgt ccgatggatc ttcagagatc ttcttcaaga
301 tcaaaaagac cactccttta agaaggctga tggaagcgtt cgctaaaaga cagggtaagg
361 aaatggactc cttaagattc ttgtacgacg gtattagaat tcaagctgat cagacccctg
421 aagatttgga catggaggat aacgatatta ttgaggctca cagagaacag attggtggga
481 tcgaggaaaa cctgtacttc caatccaata ttggaagtgg ataacggatc cgcgatcgcg
541 gcgcgccacc tggtggccgg ccggtaccac gcgtgcgcgc tgatccggct gctaacaaag
601 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
661 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc
721 cacaggacgg gtgtggtcgc catgatcgcg tagtcgatag tggctccaag tagcgaagcg
781 agcaggactg ggcggcggcc aaagcggtcg gacagtgctc cgagaacggg tgcgcataga
841 aattgcatca acgcatatag cgctagcagc acgccatagt gactggcgat gctgtcggaa
901 tggacgatat cccgcaagag gcccggcagt accggcataa ccaagcctat gcctacagca
961 tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat acacggtgcc
1021 tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc gatgataagc
1081 tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
1141 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
1201 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
1261 ataaccctga taaatgcttc aataacattg aaaaaggaag agtatgagta ttcaacattt
1321 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
1381 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
1441 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
1501 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
1561 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
1621 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
1681 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
1741 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
1801 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
1861 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
1921 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
1981 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
2041 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
2101 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
2161 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
2221 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
2281 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
2341 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
2401 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
2461 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
2521 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
2581 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
2641 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
2701 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
2761 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
2821 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
2881 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
2941 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
3001 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
3061 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
3121 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc
3181 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
3241 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
3301 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
3361 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
3421 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
3481 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
3541 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
3601 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
3661 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
3721 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga
3781 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa
3841 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt
3901 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc
3961 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa
4021 cgctgcccga gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg gatatgttct
4081 gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa ttgattggct ccaattcttg
4141 gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct
4201 ccatgcaccg cgacgcaacg cggggaggca gacaaggtat agggcggcgc ctacaatcca
4261 tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc
4321 aatgatcgaa gttaggctgg taagagccgc gagcgatcct tgaagctgtc cctgatggtc
4381 gtcatctacc tgcctggaca gcatggcctg caacgcgggc atcccgatgc cgccggaagc
4441 gagaagaatc ataatgggga aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta
4501 gcccagcgcg tcggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt
4561 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga
4621 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc
4681 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
4741 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
4801 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc
4861 gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc
4921 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
4981 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
5041 gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa
5101 t
//
