| 质粒名称: | PB-TRE3G-SOX17 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 104541 |
| 存储实验室: | David Vereide |
|
载体骨架基本信息 |
|
| 载体名称: | PiggyBac |
| 载体类型: | 哺乳动物表达，细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素, 100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
|
插入基因信息 |
|
| 插入基因名称: | SRY-box 17 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1245 bp |
| 启动子: | TRE3G |
| 克隆方法: | Gateway 克隆 |
| 5’测序引物 | GAACGTATAAGCTTTAGGCG |
|
文献引用方法 |
|
| 材料和方法部分: | PB-TRE3G-SOX17 was a gift from David Vereide (Addgene plasmid # 104541 ; http://n2t.net/addgene:104541 ; RRID:Addgene_104541) |
| 参考文献部分: | Expandable Arterial Endothelial Precursors from Human CD34(+) Cells Differ in Their Proclivity to Undergo an Endothelial-to-Mesenchymal Transition. Miller AZ, Satchie A, Tannenbaum AP, Nihal A, Thomson JA, Vereide DT. Stem Cell Reports. 2018 Jan 9;10(1):73-86. doi: 10.1016/j.stemcr.2017.12.011. 10.1016/j.stemcr.2017.12.011 PubMed 29320761 |
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 PB-TRE3G-SOX17 6425 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION PiggyBac vector encoding doxycycline-inducible human SOX17.
ACCESSION .
VERSION .
KEYWORDS PB-TRE3G-SOX17
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6425)
AUTHORS Miller AZ, Satchie A, Tannenbaum AP, Nihal A, Thomson JA, Vereide DT
TITLE Expandable Arterial Endothelial Precursors from Human CD34(+) Cells
Differ in Their Proclivity to Undergo an Endothelial-to-Mesenchymal
Transition.
JOURNAL Stem Cell Reports. 2018 Jan 9;10(1):73-86. doi:
10.1016/j.stemcr.2017.12.011.
PUBMED 29320761
REFERENCE 2 (bases 1 to 6425)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6425
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(31..53)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 153..172
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 366..388
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 380..397
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 381..397
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
repeat_region 420..482
/label=piggyBac right (3') inverted repeat
/note="piggyBac transposon-specific inverted terminal
repeat sequence (ITR)"
promoter 1430..1808
/label=TRE3G promoter
/note="3rd-generation Tet-responsive promoter that can be
activated by binding of Tet-On(R) 3G"
protein_bind 1438..1456
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 1474..1492
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 1510..1528
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 1546..1564
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 1582..1600
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 1618..1636
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 1654..1672
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 1717..1741
/label=LNCX
/note="Human CMV promoter, forward primer"
protein_bind 1826..1850
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 2816..2833
/codon_start=1
/product="HQHQHQ metal affinity tag"
/label=HQ tag
/note="The HQ tag binds to nickel and other metal ions much
like the 6xHis tag, but may allow elution at lower
concentrations of imidazole (Godat et al., 2008)."
/translation="HQHQHQ"
CDS 2822..2839
/codon_start=1
/product="HQHQHQ metal affinity tag"
/label=HQ tag
/note="The HQ tag binds to nickel and other metal ions much
like the 6xHis tag, but may allow elution at lower
concentrations of imidazole (Godat et al., 2008)."
/translation="HQHQHQ"
protein_bind complement(3129..3153)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
polyA_signal 3203..3324
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3214..3233)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 3268..3287
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
protein_bind 3795..3828
/label=lox2272
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GGATACTT). lox2272 sites are compatible with each
other, but incompatible with loxP or loxN sites (Lee and
Saito, 1988)."
repeat_region 4146..4180
/label=piggyBac left (5') inverted repeat
/note="piggyBac transposon-specific inverted terminal
repeat sequence (ITR)"
primer_bind complement(4206..4222)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4206..4222)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4219..4241)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4230..4246
/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(4254..4284)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4299..4320
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4437..4454)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4608..5196)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4688..4707)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5367..6227)
/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 5990..6009
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6228..6332)
/gene="bla"
/label=AmpR promoter
primer_bind 6400..6418
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
61 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
121 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
181 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
241 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
301 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
361 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttcaagcttt attatttaac
421 cctagaaaga taatcatatt gtgacgtacg ttaaagataa tcatgcgtaa aattgacgca
481 tgtgttttat cggtctgtat atcgaggttt atttattaat ttgaatagat attaagtttt
541 attatattta cacttacata ctaataataa attcaacaaa caatttattt atgtttattt
601 atttattaaa aaaaaacaaa aactcaaaat ttcttctata aagtaacaaa acttttaaac
661 attctctctt ttacaaaaat aaacttattt tgtactttaa aaacagtcat gttgtaatcg
721 ataataagta attagcttaa cctatacata atagaaacaa attatactta ttagtcagtc
781 agaaacaact ttggcacata tcaatattat gctctcgtta attaactcga gggatccggc
841 gcgcctgtca ttctaaatct ctctttcagc ctaaagcttt ttccccgtat ccccccaggt
901 gtctgcaggc tcaaagagca gcgagaagcg ttcagaggaa agcgatcccg tgccaccttc
961 cccgtgcccg ggctgtcccc gcacgctgcc ggctcgggga tgcgggggga gcgccggacc
1021 ggagcggagc cccgggcggc tcgctgctgc cccctagcgg gggagggacg taattacatc
1081 cctgggggct ttgggagggg gctgtccccg tgagctccca ggcgcgcctg tcattctaaa
1141 tctctctttc agcctaaagc tttttccccg tatcccccca ggtgtctgca ggctcaaaga
1201 gcagcgagaa gcgttcagag gaaagcgatc ccgtgccacc ttccccgtgc ccgggctgtc
1261 cccgcacgct gccggctcgg ggatgcgggg ggagcgccgg accggagcgg agccccgggc
1321 ggctcgctgc tgccccctag cgggggaggg acgtaattac atccctgggg gctttggggg
1381 ggggctgtcc ccgtgagctc ccaggcgcgc cgctagccaa gaattcctcg agtttactcc
1441 ctatcagtga tagagaacgt atgaagagtt tactccctat cagtgataga gaacgtatgc
1501 agactttact ccctatcagt gatagagaac gtataaggag tttactccct atcagtgata
1561 gagaacgtat gaccagttta ctccctatca gtgatagaga acgtatctac agtttactcc
1621 ctatcagtga tagagaacgt atatccagtt tactccctat cagtgataga gaacgtataa
1681 gctttaggcg tgtacggtgg gcgcctataa aagcagagct cgtttagtga accgtcagat
1741 cgcctggagc aattccacaa cacttttgtc ttataccaac tttccgtacc acttcctacc
1801 ctcgtaaacc gcggtggcgg ccatcacaag tttgtacaaa aaagcaggct tggaaggagt
1861 tcgaaccatg agcagcccgg atgcgggata cgccagtgac gaccagagcc agacccagag
1921 cgcgctgccc gcggtgatgg ccgggctggg cccctgcccc tgggccgagt cgctgagccc
1981 catcggggac atgaaggtga agggcgaggc gccggcgaac agcggagcac cggccggggc
2041 cgcgggccga gccaagggcg agtcccgtat ccggcggccg atgaacgctt tcatggtgtg
2101 ggctaaggac gagcgcaagc ggctggcgca gcagaatcca gacctgcaca acgccgagtt
2161 gagcaagatg ctgggcaagt cgtggaaggc gctgacgctg gcggagaagc ggcccttcgt
2221 ggaggaggca gagcggctgc gcgtgcagca catgcaggac caccccaact acaagtaccg
2281 gccgcggcgg cgcaagcagg tgaagcggct gaagcgggtg gagggcggct tcctgcacgg
2341 cctggctgag ccgcaggcgg ccgcgctggg ccccgagggc ggccgcgtgg ccatggacgg
2401 cctgggcctc cagttccccg agcagggctt ccccgccggc ccgccgctgc tgcctccgca
2461 catgggcggc cactaccgcg actgccagag tctgggcgcg cctccgctcg acggctaccc
2521 gttgcccacg cccgacacgt ccccgctgga cggcgtggac cccgacccgg ctttcttcgc
2581 cgccccgatg cccggggact gcccggcggc cggcacctac agctacgcgc aggtctcgga
2641 ctacgctggc cccccggagc ctcccgccgg tcccatgcac ccccgactcg gcccagagcc
2701 cgcgggtccc tcgattccgg gcctcctggc gccacccagc gcccttcacg tgtactacgg
2761 cgcgatgggc tcgcccgggg cgggcggcgg gcgcggcttc cagatgcagc cgcaacacca
2821 gcaccagcac cagcaccagc accacccccc gggccccgga cagccgtcgc cccctccgga
2881 ggcactgccc tgccgggacg gcacggaccc cagtcagccc gccgagctcc tcggggaggt
2941 ggaccgcacg gaatttgaac agtatctgca cttcgtgtgc aagcctgaga tgggcctccc
3001 ctaccagggg catgactccg gtgtgaatct ccccgacagc cacggggcca tttcctcggt
3061 ggtgtccgac gccagctccg cggtatatta ctgcaactat cctgacgtgt agctcgagtg
3121 cggccgcaac ccagctttct tgtacaaagt ggtgatccaa ccggtataga tctcgagctc
3181 gatgagttta aaccagacat gataagatac attgatgagt ttggacaaac cacaactaga
3241 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
3301 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
3361 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtatggct
3421 gattatgatc cggctgcctc gcgcttaatt aacccggggg ccgcctggga actgccgctg
3481 gccccccacc gccccaagga tctccggtcc ccgcccggct tgctgacgtc acggatgccc
3541 cagggtgtgc tgggcaggta gcgggagcct gggaaatgga gtccattagc agaagtggcc
3601 acttccagga gtcgctgtgc cccgatgcac actgggaagt ccgcagctcc gaggccccca
3661 gtggaaatcg ccagatgagg gcctcctccg gggaatgctg ggaaatggag tctacaggcc
3721 ggaggggtcc ccacggcata ctagcaagtg tgtgtagcac cgcgtaaagg ggatgaatag
3781 cagactgccc cgggataact tcgtatagga tactttatac gaagttatga gctctgtgat
3841 aatcactagt gaattgatac tagtcggtca cgatatctat aacaagaaaa tatatatata
3901 ataagttatc acgtaagtag aacatgaaat aacaatataa ttatcgtatg agttaaatct
3961 taaaagtcac gtaaaagata atcatgcgtc attttgactc acgcggtcgt tatagttcaa
4021 aatcagtgac acttaccgca ttgacaagca cgcctcacgg gagctccaag cggcgactga
4081 gatgtcctaa atgcacagcg acggattcgc gctatttaga aagagagagc aatatttcaa
4141 gaatgcatgc gtcaatttta cgcagactat ctttctaggg ttaaaaaaag cttggcgtaa
4201 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
4261 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
4321 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
4381 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
4441 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
4501 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
4561 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
4621 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
4681 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
4741 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
4801 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
4861 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
4921 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
4981 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
5041 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
5101 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
5161 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
5221 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
5281 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
5341 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
5401 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
5461 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
5521 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
5581 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
5641 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
5701 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
5761 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
5821 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
5881 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
5941 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
6001 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
6061 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
6121 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
6181 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
6241 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
6301 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
6361 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
6421 tttcg
//
