SOX17载体质粒图谱、序列、价格、抗性、测序引物

作者：杨超月

载体基本信息

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
载体出品公司: --
空载体大小: --
完整质粒大小: --
载体修饰: --
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
拷贝数: --

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
基因突变: --
GenBank编号 --
启动子: TRE3G
标签/融合蛋白: --
克隆方法: Gateway 克隆
5’克隆位点: --
3’克隆位点: --
5’测序引物 GAACGTATAAGCTTTAGGCG
3’测序引物 --

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 ; :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

载体质粒图谱和多克隆位点信息

PB-TRE3G-SOX17载体图谱

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 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 //