| 质粒名称: | pSpliceExpress |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 32485 |
| 存储实验室: | Stefan Stamm |
|
载体骨架基本信息 |
|
| 载体名称: | PET01 |
| 载体出品公司: | Mo Bi Tec |
| 载体抗性: | 氯霉素和氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | -- |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 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: | pSpliceExpress was a gift from Stefan Stamm (Addgene plasmid # 32485 ; http://n2t.net/addgene:32485 ; RRID:Addgene_32485) |
| 参考文献部分: | Rapid generation of splicing reporters with pSpliceExpress. Kishore S, Khanna A, Stamm S. Gene. 2008 Dec 31;427(1-2):104-10. Epub 2008 Oct 1. 10.1016/j.gene.2008.09.021 PubMed 18930792 |
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 pSpliceExpress 6920 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pSpliceExpress
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6920)
AUTHORS Kishore S, Khanna A, Stamm S
TITLE Rapid generation of splicing reporters with pSpliceExpress.
JOURNAL Gene. 2008 Dec 31;427(1-2):104-10. Epub 2008 Oct 1.
PUBMED 18930792
REFERENCE 2 (bases 1 to 6920)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6920
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 26..112
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 161..183
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 175..192
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 176..192
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 209..440
/gene="mutant version of attP"
/label=attP1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction
(pDONR(TM)201 version)"
CDS complement(836..1141)
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind complement(938..957)
/label=ccdB-fwd
/note="ccdB gene, forward primer"
CDS complement(1491..2144)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG"
primer_bind 2059..2078
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(2145..2247)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
protein_bind complement(2392..2623)
/gene="mutant version of attP"
/label=attP2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction
(pDONR(TM)201 version)"
primer_bind complement(2641..2660)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(2642..2660)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(2665..2681)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2665..2681)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3302..3320)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3388..3492
/gene="bla"
/label=AmpR promoter
CDS 3493..4353
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(3711..3730)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
primer_bind 4889..4908
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter complement(5463..5659)
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 5477..5612
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(5531..5550)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 5767..5786
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
terminator 6827..6854
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
ORIGIN
1 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
61 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg
121 cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa
181 acgacggcca gtcttaagct cgggccccaa ataatgattt tattttgact gatagtgacc
241 tgttcgttgc aacaaattga tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa
301 gctgaatgag aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa
361 aaacagacta catcatactg taaaacacaa catatccagt cactatgaat caactactta
421 gatggtatta gtgacctgta gtcgaccgac agccttccaa atgttcttcg ggtgatgctg
481 ccaacttagt cgaccgacag ccttccaaat gttcttctca aacggaatcg tcgtatccag
541 cctactcgct attgtcctca atgccgtatt aaatcataaa aagaaataag aaaaagaggt
601 gcgagcctct tttttgtgtg acaaaataaa aacatctacc tattcatata cgctagtgtc
661 atagtcctga aaatcatctg catcaagaac aatttcacaa ctcttatact tttctcttac
721 aagtcgttcg gcttcatctg gattttcagc ctctatactt actaaacgtg ataaagtttc
781 tgtaatttct actgtatcga cctgcagact ggctgtgtat aagggagcct gacatttata
841 ttccccagaa catcaggtta atggcgtttt tgatgtcatt ttcgcggtgg ctgagatcag
901 ccacttcttc cccgataacg gagaccggca cactggccat atcggtggtc atcatgcgcc
961 agctttcatc cccgatatgc accaccgggt aaagttcacg ggagacttta tctgacagca
1021 gacgtgcact ggccaggggg atcaccatcc gtcgcccggg cgtgtcaata atatcactct
1081 gtacatccac aaacagacga taacggctct ctcttttata ggtgtaaacc ttaaactgca
1141 tttcaccagc ccctgttctc gtcagcaaaa gagccgttca tttcaataaa ccgggcgacc
1201 tcagccatcc cttcctgatt ttccgctttc cagcgttcgg cacgcagacg acgggcttca
1261 ttctgcatgg ttgtgcttac cagaccggag atattgacat catatatgcc ttgagcaact
1321 gatagctgtc gctgtcaact gtcactgtaa tacgctgctt catagcatac ctctttttga
1381 catacttcgg gtatacatat cagtatatat tcttataccg caaaaatcag cgcgcaaata
1441 cgcatactgt tatctggctt ttagtaagcc ggatccacgc ggcgtttacg ccccccctgc
1501 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa
1561 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
1621 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
1681 ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta
1741 gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac
1801 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
1861 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
1921 atacggaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
1981 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
2041 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
2101 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
2161 cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga
2221 aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt ggcccagggc
2281 ttcccggtat caacagggac accaggattt atttattctg cgaagtgatc ttccgtcaca
2341 ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca acttagtcga ctacaggtca
2401 ctaataccat ctaagtagtt gattcatagt gactggatat gttgtgtttt acagtattat
2461 gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt
2521 tctcgttcag ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt
2581 tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc
2641 ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctc tagagcggcc
2701 gccaccgcgg tggagctcgg tacctatttg gggaccccat agagcactgc actgactgag
2761 ggatggtaac aggatgtgta ggttttggag gcccatatgt ccattcatga ccagtgactt
2821 gtctcacagc catgcaaccc ttgcctcctg tgctgactta gcaggggata aagtgagaga
2881 aagcctgggc taatcagggg gtcgctcagc tcctcctaac tggattgtcc tatgtgtctt
2941 tgcttctgtg ctgctgatgc tctgccctgt gctgacatga cctccctggc agtggcacaa
3001 ctggagctgg gtggaggccc gtgaccttca gaccttggca ctggaggtgg cccggcagaa
3061 gcgcggcatc gtggatcagt gctgcaccag catctgctct ctctaccaac tggagaacta
3121 ctgcaactag gcccaccact accctgtcca cccctctgca atgaataaaa cctttgaaag
3181 agcactacaa gttgtgtgta catgcgtgca tgtgcatatg tggtgcgggg ggaacatgag
3241 tggggctggc tggagtggcg atgataagct gtcaaacatg agaattcttg aagacgaaag
3301 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
3361 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
3421 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
3481 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
3541 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
3601 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
3661 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
3721 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
3781 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
3841 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
3901 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
3961 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
4021 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
4081 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
4141 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
4201 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
4261 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
4321 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
4381 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggctgc tgcttgcaaa
4441 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
4501 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
4561 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
4621 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
4681 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
4741 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
4801 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
4861 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
4921 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
4981 tatggaaaaa cgccagcaac ggagatgcgc cgcgtgcggc tgctggagat ggcggacgcg
5041 atggatatgt tctgccaagg gttggtttgc gcattcacag ttctccgcaa gaattgattg
5101 gctccaattc ttggagtggt gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg
5161 aggtggcccg gctccatgca ccgcgacgca acgcggggag gcagacaagg tatagggcgg
5221 cgcctacaat ccatgccaac ccgttccatg tgctcgccga ggcggcataa atcgccgtga
5281 cgatcagcgg tccaatgatc gaagttaggc tggtaagagc cgcgagcgat ccttgaagct
5341 gtccctgatg gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg ggcatcccga
5401 tgccgccgga agcgagaaga atcataatgg ggaaggccat ccagcctcgc gtcggggagc
5461 tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag
5521 gccgaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa
5581 tgggcggaac tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt
5641 gctgactaat tgagatgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
5701 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
5761 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
5821 ggccacgatg cgtccggcgt agaggatctc aggatatagt agtttcgctt ttgcataggg
5881 agggggaaat gtagtcttat gcaatactct tgtagtcttg caacatgctt atgtaacgat
5941 gagttagcaa catgccttat aaggagagaa aaagcaccgt gcatgccgat tggtgggagt
6001 aaggtggtat gatcgtggta tgatcgtgcc ttgttaggaa ggcaacagac gggtctaaca
6061 cggattggac gaaccactga attccgcatt gcagagatat tgtatttaag tgcctagctc
6121 gatacaataa acgccatttg accattcacc acattggtgt gcacctcaag cttcctgcat
6181 gctgctgctg ctgctgctgc tgggcctgag gctacagctc tccctgggca tcatcccagt
6241 tgaggaggag aacccggact tctggaaccg cgaggcagcc gaggccctgg gtgccgccaa
6301 gaagctgcag cctgcacaga cagccgccaa gaacctcatc atcttcctgg gcgatgggat
6361 gggggtgtct acggtgacag ctgccaggat cgatccgctt cctgcccctg ctggccctgc
6421 tcatcctctg ggagccccgc cctgcccagg cttttgtcaa acagcacctt tgtggttctc
6481 acttggtgga agctctctac ctggtgtgtg gggagcgtgg attcttctac acacccatgt
6541 cccgccgcga agtggaggac ccacaaggta agctctgctc ctgaattaat tctatcccaa
6601 gtgctaacta ccctgtttgt ctttcaccct tgagaccttg taaattgtgc cctaggtgtg
6661 gagggtctca ggctaaccag tggggggcac atttctgtgg gcagctagac atatgtaaac
6721 atggtagctg ccaggaagga gtgagaatcc ttccttaagt ctcctaggtg gtgacgggtg
6781 gctaggcccc aggataggta ccgggccccc cctcgagaga aacgcaaaaa ggccatccgt
6841 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
6901 ccctccgggc cgttgcttca
//
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