| 质粒名称: | pJL1-sfGFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 102634 |
| 存储实验室: | Michael Jewett |
|
载体骨架基本信息 |
|
| 载体名称: | pJL1 |
| 载体出品公司: | -- |
| 空载体大小: | 1759 bp |
| 完整质粒大小: | 2476 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 卡那霉素，50μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | sfGFP |
| 物种来源: | -- |
| 插入大小: | 723 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | T7 |
| 标签/融合蛋白: | strep标签（插入件上的C端子） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NdeI(未破坏) |
| 3’克隆位点: | SalI (未破坏) |
| 5’测序引物 | TAATACGACTCACTATAGGG |
| 3’测序引物 | TAGTTATTGCTCAGCGGTGG |
|
文献引用方法 |
|
| 材料和方法部分: | pJL1-sfGFP was a gift from Michael Jewett (Addgene plasmid # 102634 ; http://n2t.net/addgene:102634 ; RRID:Addgene_102634) |
| 参考文献部分: | BioBits Bright: A fluorescent synthetic biology education kit. Stark JC, Huang A, Nguyen PQ, Dubner RS, Hsu KJ, Ferrante TC, Anderson M, Kanapskyte A, Mucha Q, Packett JS, Patel P, Patel R, Qaq D, Zondor T, Burke J, Martinez T, Miller-Berry A, Puppala A, Reichert K, Schmid M, Brand L, Hill LR, Chellaswamy JF, Faheem N, Fetherling S, Gong E, Gonzalzles EM, Granito T, Koritsaris J, Nguyen B, Ottman S, Palffy C, Patel A, Skweres S, Slaton A, Woods T, Donghia N, Pardee K, Collins JJ, Jewett MC. Sci Adv. 2018 Aug 1;4(8):eaat5107. doi: 10.1126/sciadv.aat5107. eCollection 2018 Aug. 10.1126/sciadv.aat5107 PubMed 30083609 |
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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 pJL1-sfGFP 2486 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION In vitro expression of sfGFP from the T7 promoter.
ACCESSION .
VERSION .
KEYWORDS pJL1-sfGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2486)
AUTHORS Stark JC, Huang A, Nguyen PQ, Dubner RS, Hsu KJ, Ferrante TC,
Anderson M, Kanapskyte A, Mucha Q, Packett JS, Patel P, Patel R, Qaq
D, Zondor T, Burke J, Martinez T, Miller-Berry A, Puppala A,
Reichert K, Schmid M, Brand L, Hill LR, Chellaswamy JF, Faheem N,
Fetherling S, Gong E, Gonzalzles EM, Granito T, Koritsaris J, Nguyen
B, Ottman S, Palffy C, Patel A, Skweres S, Slaton A, Woods T,
Donghia N, Pardee K, Collins JJ, Jewett MC
TITLE BioBits Bright: A fluorescent synthetic biology education kit.
JOURNAL Sci Adv. 2018 Aug 1;4(8):eaat5107. doi: 10.1126/sciadv.aat5107.
eCollection 2018 Aug.
PUBMED 30083609
REFERENCE 2 (bases 1 to 2486)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..2486
/organism="synthetic DNA construct"
/mol_type="other DNA"
RBS 113..135
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
primer_bind complement(166..185)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(167..185)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin complement(213..801)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(293..312)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind complement(912..931)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 923..1738
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(996..1015)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
terminator 1756..1803
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 1799..1817
/label=T7 Term
/note="T7 terminator, reverse primer"
CDS complement(1872..1895)
/codon_start=1
/product="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
/label=Strep-Tag II
/translation="WSHPQFEK"
CDS complement(1896..1943)
/codon_start=1
/product="11th beta-strand of superfolder GFP"
/label=GFP11
/note="associates with GFP(1-10), consisting of the first
10 beta-strands, to reconstitute fluorescent GFP (Cabantous
et al., 2005; Kamiyama et al., 2016)"
/translation="RDHMVLHEYVNAAGIT"
ORIGIN
1 gccttcacct tcaccacgca cgctgaattt gtgaccgttc acatcgccat ccagttccac
61 cagaatcggc acaacgccgg taaacagttc ttcacctttg ctcatatgta tatctccttc
121 ttaaagttaa acaaaattat ttctagaggg aaaccgttgt ggtctcccta tagtgagtcg
181 tattaatttc gcgggatcgc gttgctggcg tttttccata ggctccgccc ccctgacgag
241 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
301 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
361 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
421 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
481 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
541 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
601 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
661 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
721 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
781 cgcagaaaaa aaggatctca agaagatcct ttgatctcac gttgtgtctc aaaatctctg
841 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
901 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctcgaagcc
961 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
1021 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt
1081 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
1141 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
1201 tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata
1261 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
1321 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
1381 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
1441 gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt
1501 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
1561 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
1621 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
1681 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
1741 agaattggct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc
1801 tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt tgttagcagc
1861 cggtcgactt atttttcgaa ctgcggatgg ctccacgtaa tacctgccgc attcacatat
1921 tcgtgcagaa ccatgtggtc ccgcgtgcct ttttcgttcg gatctttaga cagaacggtc
1981 tgcgtgctca gataatgatt atccggcagc agcaccggac catcaccaat cggggtattc
2041 tgctgataat gatctgccag ctgcacactg ccatcttcaa cgttatggcg aacggtaaaa
2101 ttcgctttga tgccattttt ctgtttatcc gccgtaatat aaacattatg gctattaaag
2161 ttgtattcca gtttatggcc caggatattg ccatcttctt taaaatccgt gcctttcagt
2221 tcaatgcggt tcaccagggt atcgccttca aatttcacaa cggcgcgcgt tttatatttg
2281 ccatcatctt taaagctaat cgtacgttcc tgcacatagc cttccggcat tgcagattta
2341 aagaaatcgt gacgtttcat gtgatccgga tagcgactaa aacactgaac gccataggtc
2401 agggtggtca ccagcgtcgg ccacggcacc ggcagtttgc cggtggtgca gataaatttc
2461 agcgtcagtt tgccaatcgt ggcatc
//
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