| 质粒名称: | pBAD_sfGFP3TCG4TAG |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 174518 |
| 存储实验室: | Jason W Chin |
|
载体骨架基本信息 |
|
| 载体名称: | pBAD |
| 空载体大小: | 3896bp |
| 完整质粒大小: | 4646bp |
| 载体类型: | 细菌细胞表达 |
| 载体抗性: | 阿泊拉霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | sfGFP |
| 插入大小: | 750bp |
| 启动子: | araBAD |
| 标签/融合蛋白: | 6 x His (C terminal on insert) |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | ATGCCATAGCATTTTTATCC |
| 3’测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: |
pBAD_sfGFP3TCG4TAG was a gift from Jason W Chin (Addgene plasmid # 174518 ; http://n2t.net/addgene:174518 ; RRID:Addgene_174518)
|
| 参考文献部分: | Sense codon reassignment enables viral resistance and encoded polymer synthesis. Robertson WE, Funke LFH, de la Torre D, Fredens J, Elliott TS, Spinck M, Christova Y, Cervettini D, Boge FL, Liu KC, Buse S, Maslen S, Salmond GPC, Chin JW. Science. 2021 Jun 4;372(6546):1057-1062. doi: 10.1126/science.abg3029. 10.1126/science.abg3029 PubMed 34083482 |
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 pBAD_sfGFP3TCG4TAG 4646 bp ds-DNA circular SYN 22-SEP-2021
DEFINITION Recoded sfGFP reporter with the codons TCG and TAG inserted at
positions 3 and 4, respectively..
ACCESSION .
VERSION .
KEYWORDS pBAD_sfGFP3TCG4TAG
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4646)
AUTHORS Robertson WE, Funke LFH, de la Torre D, Fredens J, Elliott TS,
Spinck M, Christova Y, Cervettini D, Boge FL, Liu KC, Buse S, Maslen
S, Salmond GPC, Chin JW
TITLE Sense codon reassignment enables viral resistance and encoded
polymer synthesis.
JOURNAL Science. 2021 Jun 4;372(6546):1057-1062. doi:
10.1126/science.abg3029.
PUBMED 34083482
REFERENCE 2 (bases 1 to 4646)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 22, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4646
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(17..34)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 187..273
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind complement(480..497)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(614..1159)
/direction=LEFT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
"
primer_bind complement(694..713)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 1271..1299
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
promoter 1347..1438
/gene="bla"
/label=AmpR promoter
primer_bind 2372..2394
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
promoter 3455..3739
/gene="araBAD"
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
primer_bind 3662..3681
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
CDS 3785..4495
/codon_start=1
/product="GFP variant that folds robustly even when fused
to poorly folded proteins (Pedelacq et al., 2006)"
/label=superfolder GFP
/note="mammalian codon-optimized"
/translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
VTAAGITHGMDELYK"
CDS 4502..4519
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 4545..4574
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 4590..4607
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
ORIGIN
1 gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac
61 agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag
121 tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
181 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
241 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagctgtccc tcctgttcag
301 ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg acatcagcgc tagcggagtg
361 tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc atgtggcagg
421 agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata ttccgcttcc
481 tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa
541 cggggcggag atttcctgga agatgccagg aagatactta acagggaagt gagagggccg
601 cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct
661 caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg
721 gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat
781 ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag
841 ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat
901 cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat
961 tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag
1021 ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag
1081 aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agtttacgcg
1141 cagaccaaaa cgatctcaag aagatcatct tattaatcag ataaaatatt tctagatttc
1201 agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgta
1261 attctcatgt ttgacagctt atcatcgata agctttaatg cggtagttta tcacagttaa
1321 attgctaacg cagtcaggca ccgtgttttg tttatttttc taaatacatt caaatatgta
1381 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
1441 gagtagtgcg gtggagtgca atgtcgtgca atacgaatgg cgaaaagccg agctcatcgg
1501 tcagcttctc aaccttgggg ttacccccgg cggtgtgctg ctggtccaca gctccttccg
1561 tagcgtccgg cccctcgaag atgggccact tggactgatc gaggccctgc gtgctgcgct
1621 gggtccggga gggacgctcg tcatgcccag ctggagtggt ctggacgacg agccgttcga
1681 tcctgccacg agccccgtta caccggacct tggagttgtc tctgacacat tctggcgcct
1741 gccaaatgta aagcgcagcg cccatccatt tgcctttgcg gcagcggggc cacaggcaga
1801 gcagatcatc tctgatccat tgcccctgcc acctcacagc cctgcaagcc cggtcgcccg
1861 tgtccatgaa ctcgatgggc aggtacttct cctcggcgtg ggacacgatg ccaacacgac
1921 gctgcatctt gccgagttga tggcaaaggt tccctatggg gtgccgagac actgcaccat
1981 tcttcaggat ggcaagttgg tacgcgtcga ttatctcgag aatgaccact gctgtgagcg
2041 ctttgccttg gcggacaggt ggctcaagga gaagagcctt cagaaggaag gtccagtcgg
2101 tcatgccttt gctcggttga tccgctcccg cgacattgtg gcgacagccc tgggtcaact
2161 gggccgagat ccgttgatct tcctgcatcc gccagaggcg ggatgcgaag aatgcgatgc
2221 cgctcgccag agcattggct gaatggaagc cggcggcacc tcgctaacgg attcaccact
2281 ccaagaattg gagccaatca attcttgcgg agaactgtga atgcgcaaac caacccttgg
2341 cagaacatat ccatcgcgtg cggccgcgtc tccgggagct gcatgtgtca gaggttttca
2401 ccgtcatcac cgaaacgcgc gaggcagcag atcaattcgc gcgcgaaggc gaagcggcat
2461 gcataatgtg cctgtcaaat ggacgaagca gggattctgc aaaccctatg ctactccgtc
2521 aagccgtcaa ttgtctgatt cgttaccaat tagctcaact tgacggctac atcattcact
2581 ttttcttcac aaccggcacg gaactcgctc gggctggccc cggtgcattt tttaaatacc
2641 cggctgaaat agagttgatc gtcaaaacca acattgcgac cgacggtggc gataggcatc
2701 cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag
2761 acgctaatcc ctaactgctg gcggaaaaga tgactcagac ggctcgggct caagcaaaca
2821 tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct gatgtactga
2881 caagcctcgc gtacccgatt atccatcggt ggatggaggc tctcgttaat cgcttccatg
2941 cgccgcagta acaattgctc aagcagattt atcgccagca gctcgctata gcgcccttcc
3001 ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca
3061 tccgggcgaa agaaccccgt attggcaaat atactcggcc agttaagcca ttcatgccag
3121 taggcgcgcg gacgaaagta aacccactgg tgataccatt cgcgagcctc cggatgacga
3181 ccgtagtgat gaatctctcc tggcgggaac agcaaaatat cacccggtcg gcaaacaaat
3241 tctcgtccct gatttttcac caccccctga ccgcgaatgg tgagattgag aatataacct
3301 ttcattccca gcggtcggtc gataaaaaaa tcgagataac cgttggcctc aatcggcgtt
3361 aaacccgcca ccagatgggc attaaagctg tatcccggca gcaggggatc attttgcgct
3421 tcagccatac ttttcatact cccgccattc agagaagaaa ccaattgtcc atattgcatc
3481 agacattgcc gtcactgcgt cttttactgg ctcttctcgc taaccaaacc ggtaaccccg
3541 cttattaaaa gcattctgta acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa
3601 agtgtctata atcacggcag aaaagtccac attgattatt tgcacggcgt cacactttgc
3661 tatgccatag catttttatc cataagatta gcggatccta cctgacgctt tttatcgcaa
3721 ctctctactg tttctccata cccgtttttt gggctaacag gaggaattaa ccatggcttc
3781 gtagagcaaa ggtgaagaac tgtttaccgg cgttgtgccg attctggtgg aactggatgg
3841 tgatgtgaat ggccataaat ttagcgttcg tggcgaaggc gaaggtgatg cgaccaacgg
3901 taaactgacc ctgaaattta tttgcaccac cggtaaactg ccggttccgt ggccgaccct
3961 ggtgaccacc ctgacctatg gcgttcagtg ctttagccgc tatccggatc atatgaaacg
4021 ccatgatttc tttaaaagcg cgatgccgga aggctatgtg caggaacgta ccattagctt
4081 caaagatgat ggcacctata aaacccgtgc ggaagttaaa tttgaaggcg ataccctggt
4141 gaaccgcatt gaactgaaag gtattgattt taaagaagat ggcaacattc tgggtcataa
4201 actggaatat aatttcaaca gccataacgt gtatattacc gccgataaac agaaaaatgg
4261 catcaaagcg aactttaaaa tccgtcacaa cgtggaagat ggtagcgtgc agctggcgga
4321 tcattatcag cagaataccc cgattggtga tggcccggtg ctgctgccgg ataatcatta
4381 tctgagcacc cagagcgttc tgagcaaaga tccgaatgaa aaacgtgatc atatggtgct
4441 gctggaattt gttaccgccg cgggcattac ccacggtatg gatgaactgt ataaaggcag
4501 ccaccatcat catcaccatt aaagctcgag cgaagcttgg gcccgaacaa aaactcatct
4561 cagaagagga tctgaatagc gccgtcgacc atcatcatca tcatcattga gtttaaacgg
4621 tctccagctt ggctgttttg gcggat
//
