| 质粒名称: | pST1374-scFv-APOBEC-UGI-GB1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 113029 |
| 存储实验室: | Xingxu Huang |
|
载体骨架基本信息 |
|
| 载体名称: | pST1374 |
| 载体出品公司: | -- |
| 空载体大小: | 4968 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素 |
| 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: | scFv-rAPOBEC1-UGI-GB1,NLS-scFv-rAPOBEC1-UGI-GB1-NLS |
| 物种来源: | R. norvegicus (rat); S. pyogenes |
| 插入大小: | 2268 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | CMV |
| 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 | CMV-fwd |
| 3’测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: | pST1374-scFv-APOBEC-UGI-GB1 was a gift from Xingxu Huang (Addgene plasmid # 113029 ; http://n2t.net/addgene:113029 ; RRID:Addgene_113029) |
| 参考文献部分: | BE-PLUS: a new base editing tool with broadened editing window and enhanced fidelity. Jiang W, Feng S, Huang S, Yu W, Li G, Yang G, Liu Y, Zhang Y, Zhang L, Hou Y, Chen J, Chen J, Huang X. Cell Res. 2018 Jun 6. pii: 10.1038/s41422-018-0052-4. doi: 10.1038/s41422-018-0052-4. 10.1038/s41422-018-0052-4 [pii] PubMed 29875396 |
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 pST1374-scFv-APOBEC-UGI-GB1 7235 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses scFv-rAPOBEC1-UGI in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pST1374-scFv-APOBEC-UGI-GB1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7235)
AUTHORS Jiang W, Feng S, Huang S, Yu W, Li G, Yang G, Liu Y, Zhang Y, Zhang
L, Hou Y, Chen J, Chen J, Huang X
TITLE BE-PLUS: a new base editing tool with broadened editing window and
enhanced fidelity.
JOURNAL Cell Res. 2018 Jun 6. pii: 10.1038/s41422-018-0052-4. doi:
10.1038/s41422-018-0052-4.
PUBMED 29875396
REFERENCE 2 (bases 1 to 7235)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7235
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 11..32
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(151..171)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 156..485
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 336..471
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 398..417
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 533..580
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 599..997
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
polyA_signal 1155..1276
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1192..1211)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1246..1265
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(1325..1341)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1325..1341)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1338..1360)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1349..1365
/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(1373..1403)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1418..1439
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1556..1573)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1727..2315)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1807..1826)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2486..3346)
/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 3109..3128
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3347..3451)
/gene="bla"
/label=AmpR promoter
primer_bind complement(3526..3545)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 3717..4096
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 4097..4300
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 4251..4271
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 4345..4364
/label=T7
/note="T7 promoter, forward primer"
promoter 4345..4363
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 4392..4412
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 4422..4445
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 5193..5219
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 5331..6017
/codon_start=1
/product="cytidine deaminase (C to U editing enzyme) from
rat"
/label=APOBEC-1
/note="can use ssDNA as a substrate (Komor et al., 2016)"
/translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
LWATGLK"
CDS 6033..6281
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 6339..6500
/codon_start=1
/product="B1 domain of Streptococcal protein G "
/label=GB1
/note="effective as a solubilizing fusion partner (Cheng
and Patel, 2004)"
/translation="YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDD
ATKTFTVTE"
CDS 6528..6548
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6582..6623
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 6633..6650
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(6673..6690)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6679..6903
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 6949..142
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7036..7055)
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
61 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
121 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
181 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
241 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
301 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
361 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
421 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
481 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa
541 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat
601 ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa
661 cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg
721 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt
781 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa
841 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca
901 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg
961 tgaattgctg ccctctggtt atgtgtggga gggctaagca cttcgtggcc gaggagcagg
1021 actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
1081 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
1141 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
1201 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
1261 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
1321 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
1381 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
1441 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
1501 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
1561 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
1621 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
1681 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
1741 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
1801 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
1861 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
1921 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
1981 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
2041 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
2101 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
2161 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
2221 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
2281 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
2341 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
2401 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
2461 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
2521 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
2581 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
2641 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
2701 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
2761 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
2821 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
2881 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
2941 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
3001 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
3061 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
3121 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
3181 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
3241 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
3301 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
3361 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
3421 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
3481 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
3541 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
3601 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
3661 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
3721 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
3781 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
3841 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
3901 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
3961 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
4021 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
4081 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
4141 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
4201 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
4261 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
4321 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
4381 gcaccatggg acctaagaaa aagaggaagg tggcggccgc tgactacaag gatgacgacg
4441 ataaatctag aatgggtccc gacatcgtga tgacccagag ccccagcagc ctgagcgcca
4501 gcgtgggcga ccgcgtgacc atcacctgcc gcagcagcac cggcgccgtg accaccagca
4561 actacgccag ctgggtgcag gagaagcccg gcaagctgtt caagggcctg atcggcggca
4621 ccaacaaccg cgcccccggc gtgcccagcc gcttcagcgg cagcctgatc ggcgacaagg
4681 ccaccctgac catcagcagc ctgcagcccg aggacttcgc cacctacttc tgcgccctgt
4741 ggtacagcaa ccactgggtg ttcggccagg gcaccaaggt ggagctgaag cgcggcggcg
4801 gcggcagcgg cggcggcggc agcggcggcg gcggcagcag cggcggcggc agcgaggtga
4861 agctgctgga gagcggcggc ggcctggtgc agcccggcgg cagcctgaag ctgagctgcg
4921 ccgtgagcgg cttcagcctg accgactacg gcgtgaactg ggtgcgccag gcccccggcc
4981 gcggcctgga gtggatcggc gtgatctggg gcgacggcat caccgactac aacagcgccc
5041 tgaaggaccg cttcatcatc agcaaggaca acggcaagaa caccgtgtac ctgcagatga
5101 gcaaggtgcg cagcgacgac accgccctgt actactgcgt gaccggcctg ttcgactact
5161 ggggccaggg caccctggtg accgtgagca gctacccata cgatgttcca gattacgctg
5221 gtggaggcgg aggttctggg ggaggaggta gtggcggtgg tggttcagga ggcggcggat
5281 ccagcggcag cgagactccc gggacctcag agtccgccac acccgaaagt atgagctcag
5341 agactggccc agtggctgtg gaccccacat tgagacggcg gatcgagccc catgagtttg
5401 aggtattctt cgatccgaga gagctccgca aggagacctg cctgctttac gaaattaatt
5461 gggggggccg gcactccatt tggcgacata catcacagaa cactaacaag cacgtcgaag
5521 tcaacttcat cgagaagttc acgacagaaa gatatttctg tccgaacaca aggtgcagca
5581 ttacctggtt tctcagctgg agcccatgcg gcgaatgtag tagggccatc actgaattcc
5641 tgtcaaggta tccccacgtc actctgttta tttacatcgc aaggctgtac caccacgctg
5701 acccccgcaa tcgacaaggc ctgcgggatt tgatctcttc aggtgtgact atccaaatta
5761 tgactgagca ggagtcagga tactgctgga gaaactttgt gaattatagc ccgagtaatg
5821 aagcccactg gcctaggtat ccccatctgt gggtacgact gtacgttctt gaactgtact
5881 gcatcatact gggcctgcct ccttgtctca acattctgag aaggaagcag ccacagctga
5941 cattctttac catcgctctt cagtcttgtc attaccagcg actgccccca cacattctct
6001 gggccaccgg gttgaaaggc ggaggtggaa gcactaatct gtcagatatt attgaaaagg
6061 agaccggaaa gcaactggtt atccaggaat ccatcctcat gctcccagag gaggtggaag
6121 aagtcattgg gaacaagccg gaaagcgata tactcgtgca caccgcctac gacgagagca
6181 ccgacgagaa tgtcatgctt ctgactagcg acgcccctga atacaagcct tgggctctgg
6241 tcatacagga tagcaacggt gagaacaaga ttaagatgct cggaggagga ggaagcggag
6301 gaggaggtag cggaggaggt ggaagccgga ccgaagagta caagcttatc ctgaacggta
6361 aaaccctgaa aggtgaaacc accaccgaag ctgttgacgc tgctaccgcg gaaaaagttt
6421 tcaaacagta cgctaacgac aacggtgttg acggtgaatg gacctacgac gacgctacca
6481 aaaccttcac ggtaaccgaa ggtggtggta gcggtggtgg tggtagtccc aagaagaaga
6541 ggaaagtctc gagcggtgga gctgcaggag ggcccttcga aggtaagcct atccctaacc
6601 ctctcctcgg tctcgattct acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa
6661 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
6721 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
6781 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
6841 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
6901 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
6961 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
7021 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
7081 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
7141 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
7201 agacggtttt tcgccctttg acgttggagt ccacg
//
