| 质粒名称: | pJC53.2 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 26536 |
| 存储实验室: | Phillip Newmark |
|
载体骨架基本信息 |
|
| 载体名称: | pJC53.2 |
| 空载体大小: | 3240 bp |
| 完整质粒大小: | 4839 bp |
| 载体修饰: | T/A Cloning Vector for dsRNA generation and the generation of sense and anti-sense probes |
| 载体类型: | T/A克隆细胞表达 |
| 载体抗性: | Chloramphenicol and Kanamycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | DB3.1 |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | None |
| 插入大小: | 1600bp |
| 克隆方法: | 限制性内切酶 |
| 5’测序引物 | T3 |
| 3’测序引物 | SP6 |
|
文献引用方法 |
|
| 材料和方法部分: | pJC53.2 was a gift from Phillip Newmark (Addgene plasmid # 26536 ; http://n2t.net/addgene:26536 ; RRID:Addgene_26536) |
| 参考文献部分: | Genome-wide analyses reveal a role for peptide hormones in planarian germline development. Collins JJ, Hou X, Romanova EV, Lambrus BG, Miller CM, Saberi A, Sweedler JV, Newmark PA. PLoS Biol. 2010 . 8(10):e1000509. 10.1371/journal.pbio.100050 |
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 pJC53.2 4839 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION T/A Cloning Vector for dsRNA generation and the generation of sense
and anti-sense probes.
ACCESSION .
VERSION .
KEYWORDS pJC53.2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4839)
AUTHORS Collins JJ, Hou X, Romanova EV, Lambrus BG, Miller CM, Saberi A,
Sweedler JV, Newmark PA
TITLE Genome-wide analyses reveal a role for peptide hormones in planarian
germline development.
JOURNAL PLoS Biol. 2010 . 8(10):e1000509.
PUBMED 20967238
REFERENCE 2 (bases 1 to 4839)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4839
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 2..49
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter 63..81
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 85..103
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 237..339
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 340..993
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG"
primer_bind complement(406..425)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
CDS 1343..1648
/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 1527..1546
/label=ccdB-fwd
/note="ccdB gene, forward primer"
promoter complement(1719..1737)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1720..1737)
/label=SP6
/note="SP6 promoter, forward primer"
promoter complement(1741..1759)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
terminator 1773..1820
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind complement(1832..1849)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2003..2591)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2083..2102)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2762..3622)
/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 3385..3404
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
CDS complement(3640..4434)
/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(3751..3770)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 4361..4380
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
ORIGIN
1 tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagg cgcgccggat
61 cctaatacga ctcactatag ggagaattaa ccctcactaa agggagacct taggtcgcgg
121 ccgctttgcg ccgaataaat acctgtgacg gaagatcact tcgcagaata aataaatcct
181 ggtgtccctg ttgataccgg gaagccctgg gccaactttt ggcgaaaatg agacgttgat
241 cggcacgtaa gaggttccaa ctttcaccat aatgaaataa gatcactacc gggcgtattt
301 tttgagttat cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
361 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
421 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
481 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
541 aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat atgggatagt
601 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
661 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
721 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
781 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
841 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
901 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
961 ttacaacagt actgcgatga gtggcagggg gggcgtaaac gccgcgtgga tccggcttac
1021 taaaagccag ataacagtat gcgtatttgc gcgctgattt ttgcggtata agaatatata
1081 ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct atgaagcagc gtattacagt
1141 gacagttgac agcgacagct atcagttgct caaggcatat atgatgtcaa tatctccggt
1201 ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc gtgccgaacg ctggaaagcg
1261 gaaaatcagg aagggatggc tgaggtcgcc cggtttattg aaatgaacgg ctcttttgct
1321 gacgagaaca ggggctggtg aaatgcagtt taaggtttac acctataaaa gagagagccg
1381 ttatcgtctg tttgtggatg tacagagtga tattattgac acgcccgggc gacggatggt
1441 gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac tttacccggt
1501 ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca gtgtgccggt
1561 ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc
1621 cattaacctg atgttctggg gaatataaat gtcaggctcc cttatacaca gccagtctgc
1681 aggtcgatac agtagaaact cgaggaccta aggtcacttc tatagtgtca cctaaatctc
1741 cctatagtga gtcgtattag gatccggcgc gcctagcata accccttggg gcctctaaac
1801 gggtcttgag gggttttttg acgcgtgctt cctcgctcac tgactcgctg cgctcggtcg
1861 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
1921 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
1981 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
2041 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
2101 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
2161 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
2221 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
2281 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
2341 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
2401 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
2461 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
2521 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
2581 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
2641 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
2701 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
2761 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
2821 tagttgcctg gctccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
2881 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
2941 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
3001 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
3061 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
3121 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
3181 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
3241 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
3301 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
3361 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
3421 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
3481 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
3541 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
3601 cacggaaatg ttgaatactc atactcttcc tttttcaatt cagaagaact cgtcaagaag
3661 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg
3721 gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg
3781 atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc
3841 caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg
3901 catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc
3961 cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg
4021 tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc
4081 atcagccatg atggatactt tctcggcagg agcaaggtgg gatgacagga gatcctgccc
4141 cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc
4201 tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc
4261 attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag
4321 ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag
4381 cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa
4441 cgatcctcat cctgtctctt gatcagatct tgatcccctg cgccatcaga tccttggcgg
4501 caagaaagcc atccagttta ctttgcaggg cttcccaacc ttaccagagg gcgccccagc
4561 tggcaattcc ggttcgcttg ctgtccataa aaccgcccag tctagctatc gccatgtaag
4621 cccactgcaa gctacctgct ttctctttgc gcttgcgttt tcccttgtcc agatagccca
4681 gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtgttccg
4741 cttcctttag cagcccttgc gccctgaatt ttgttaaaat tcgcgttaaa tttttgttaa
4801 atcagctcat tttttaacca ataggccgaa atcgacgcg
//
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