| 质粒名称: | pSC101-Donor |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 140630 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pUC19, temperature sensitive pSC101 origin of replication |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | 4133 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达，CRISPR；Transposon |
| 筛选标记: | -- |
| 载体抗性: | 卡那霉素，50μg/mL |
| 生长温度: | 30°C |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | LE, RE, KanR, temperature sensitive pSC101 origin of replication |
| 物种来源: | Scytonema hofmanni |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pSC101-Donor was a gift from Sheng Yang (Addgene plasmid # 140630 ; http://n2t.net/addgene:140630 ; RRID:Addgene_140630) |
| 参考文献部分: | Multicopy chromosomal integration using CRISPR-associated transposases. Zhang Y, Sun X, Wang Q, Xu J, Dong F, Yang S, Yang J, Zhang Z, Qian Y, Chen J, Zhang J, Liu Y, Tao R, Jiang Y, Yang J, Yang S. ACS Synth Biol. 2020 Jun 17. doi: 10.1021/acssynbio.0c00073. 10.1021/acssynbio.0c00073 PubMed 32551502 |
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 pSC101-Donor 4133 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Kanamycin resistance donor for ShCAST_harboring the temperature
sensitive pSC101 origin of replication..
ACCESSION .
VERSION .
KEYWORDS pSC101-Donor
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4133)
AUTHORS Zhang Y, Sun X, Wang Q, Xu J, Dong F, Yang S, Yang J, Zhang Z, Qian
Y, Chen J, Zhang J, Liu Y, Tao R, Jiang Y, Yang J, Yang S
TITLE Multicopy chromosomal integration using CRISPR-associated
transposases.
JOURNAL ACS Synth Biol. 2020 Jun 17. doi: 10.1021/acssynbio.0c00073.
PUBMED 32551502
REFERENCE 2 (bases 1 to 4133)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4133
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind complement(549..596)
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS 983..1777
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(1037..1056)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 1647..1666
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
protein_bind complement(1885..1932)
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
terminator 1980..2066
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2169..2263
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(2580..3530)
/codon_start=1
/gene="rep101 encoding an A56V mutation"
/product="temperature-sensitive version of the RepA protein
needed for replication with the pSC101 origin (Armstrong et
al., 1984)"
/label=Rep101(Ts)
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
rep_origin 3578..3800
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
ORIGIN
1 ggtctgctat gtggtgctat ctgacttttt gctgttcagc agttcctgcc ctctgatttt
61 ccagtctgac cacttcggat tatcccgtga caggtcattc agactggcta atgcacccag
121 taaggcagcg gtatcatcaa cggggtctga cgctcagtgg aacgaaaact cacgttaagg
181 gattttggtc atgagattat caaaaaggat cttcacctag atccgttatg cagcggaaag
241 taaaaaattt ttagtttatt agacatctcc acaaaaggcg tagtgtacag tgacaaatta
301 tctgtcgtcg gtgacagatt aatgtcattg tgactattta attgtcgtcg tgacccatca
361 gcgttgctta attaattgat gacaaattaa atgtcatcaa tataatatgc tctgcaatta
421 ttatacaaag caattaaaac aagcggataa aaggacttgc tttcaaccca cccctaagtt
481 taatagttac tgagggggat ccactagtga gctcatgcat gatctcgaat tagcttcaaa
541 agcgctctga agttcctata ctttctagag aataggaact tcggaatagg aacttcaaga
601 tcccctgatt ccctttgtca acagcaatgg ataattcgat ttaacaaatg catggcgcaa
661 gggctgctaa aggaagcgga acacgtagaa agccagtccg cagaaacggt gctgaccccg
721 gatgaatgtc agctactggg ctatctggac aagggaaaac gcaagcgcaa agagaaagca
781 ggtagcttgc agtgggctta catggcgata gctagactgg gcggttttat ggacagcaag
841 cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa
901 ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagat ctgatcaaga
961 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
1021 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
1081 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
1141 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
1201 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
1261 attgggcgaa gtgccggggc aggatctcct gtcatcccac cttgctcctg ccgagaaagt
1321 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
1381 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
1441 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
1501 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
1561 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
1621 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
1681 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
1741 catcgccttc tatcgccttc ttgacgagtt cttctgaatt gaaaaaggaa gagtatgagg
1801 atccaacatt tccaatcact agtgaattat ctagaattat tccattgagt aagtttttaa
1861 gcacatcagc ttcaaaagcg ctctgaagtt cctatacttt ctagagaata ggaacttcgg
1921 aataggtact tcaagatccc caattcgaga tcgtccgggc cgcaagctcc tagcggcgga
1981 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
2041 ttcgactgag cctttcgttt tatttgatgc ctcaagctag agagtcatta ccccaggcgt
2101 ttaagggcac caataactgc cttaaaaaaa ttacgccccg ccctgccact catcgcagtc
2161 tagcttggat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc
2221 agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagctc agctaattaa
2281 ggcgacagtc aatttgtcat tatgaaaata cacaaaagct ttttcctatc ttgcaaagcg
2341 acagctaatt tgtcacaatc acggacaacg acatctattt tgtcactgca aagaggttat
2401 gctaaaactg ccaaagcgct ataatctata ctgtataagg attttactga tgacaataat
2461 ttgtcacaac gacatataat tagtcactgt acacgtagag acgtagcaat gctacctctc
2521 tacaatggtt ttgtgttagt cttgatgctt cactgataga tacaagagcc ataagaacct
2581 cagatccttc cgtatttagc cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc
2641 catgagaacg aaccattgag atcatactta ctttgcatgt cactcaaaaa ttttgcctca
2701 aaactggtga gctgaatttt tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta
2761 tgtaggtagg aatctgatgt aatggttgtt ggtattttgt caccattcat ttttatctgg
2821 ttgttctcaa gttcggttac gagatccatt tgtctatcta gttcaacttg gaaaatcaac
2881 gtatcagtcg ggcggcctcg cttatcaacc accaatttca tattgctgta agtgtttaaa
2941 tctttactta ttggtttcaa aacccattgg ttaagccttt taaactcatg gtagttattt
3001 tcaagcatta acatgaactt aaattcatca aggctaatct ctatatttgc cttgtgagtt
3061 ttcttttgtg ttagttcttt taataaccac tcataaatcc tcatagagta tttgttttca
3121 aaagacttaa catgttccag attatatttt atgaattttt ttaactggaa aagataaggc
3181 aatatctctt cactaaaaac taattctaat ttttcgcttg agaacttggc atagtttgtc
3241 cactggaaaa tctcaaagcc tttaaccaaa ggattcctga tttccacagt tctcgtcatc
3301 agctctctgg ttgctttagc taatacacca taagcatttt ccctactgat gttcatcatc
3361 tgaacgtatt ggttataagt gaacgatacc gtccgttctt tccttgtagg gttttcaatc
3421 gtggggttga gtagtgccac acagcataaa attagcttgg tttcatgctc cgttaagtca
3481 tagcgactaa tcgctagttc atttgctttg aaaacaacta attcagacat acatctcaat
3541 tggtctaggt gattttaatc actataccaa ttgagatggg ctagtcaatg ataattacta
3601 gtccttttcc tttgagttgt gggtatctgt aaattctgct agacctttgc tggaaaactt
3661 gtaaattctg ctagaccctc tgtaaattcc gctagacctt tgtgtgtttt ttttgtttat
3721 attcaagtgg ttataattta tagaataaag aaagaataaa aaaagataaa aagaatagat
3781 cccagccctg tgtataactc actactttag tcagttccgc agtattacaa aaggatgtcg
3841 caaacgctgt ttgctcctct acaaaacaga ccttaaaacc ctaaaggctt aagtagcacc
3901 ctcgcaagct cggttgcggc cgcaatcggg caaatcgctg aatattcctt ttgtctccga
3961 ccatcaggca cctgagtcgc tgtctttttc gtgacattca gttcgctgcg ctcacggctc
4021 tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt atggattcat gcaaggaaac
4081 tacccataat acaagaaaag cccgtcacgg gcttctcagg gcgttttatg gcg
//
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