| 质粒名称: | pCRZeroT |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 120276 |
| 存储实验室: | Ken Motohashi |
|
载体骨架基本信息 |
|
| 载体名称: | pUC18 |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 2875bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | ccdB Survival |
| 拷贝数: | 高拷贝 |
|
文献引用方法 |
|
| 材料和方法部分: | pCRZeroT was a gift from Ken Motohashi (Addgene plasmid # 120276 ; http://n2t.net/addgene:120276 ; RRID:Addgene_120276) |
| 参考文献部分: | A novel series of high-efficiency vectors for TA cloning and blunt-end cloning of PCR products. Motohashi K. Sci Rep. 2019 Apr 23;9(1):6417. doi: 10.1038/s41598-019-42868-6. 10.1038/s41598-019-42868-6 PubMed 31015513 |
买家导航
pCRZeroT载体质粒基本信息
pCRZeroT质粒图谱载体图谱和pCRZeroT载体序列质粒序列多克隆位点信息
pCRZeroT质粒载体简介
pCRZeroT质粒序列载体序列
LOCUS pCRZeroT 2975 bp DNA circular SYN 14-APR-2025
DEFINITION Bifunctional vector for TA-cloning and blunt-end cloning of
PCR-products using ZERO-backgound system.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2975)
AUTHORS Motohashi K
TITLE A novel series of high-efficiency vectors for TA cloning and
blunt-end cloning of PCR products.
JOURNAL Sci Rep. 2019 Apr 23;9(1):6417. doi: 10.1038/s41598-019-42868-6.
PUBMED 31015513
REFERENCE 2 (bases 1 to 2975)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep.";
date: "2019-04-23"; volume: "9(1):6417. doi"; pages: "
10.1038/s41598-019-42868-6"
COMMENT Sequence Label: pCRZeroT
FEATURES Location/Qualifiers
source 1..2975
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(10..28)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 96..200
/gene="bla"
/label=AmpR promoter
CDS join(201..269,270..1061)
/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 complement(419..438)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1232..1820
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1721..1740
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1974..1991
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 2108..2129
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter join(2144..2149,2150..2167,2168..2174)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2182..2198
/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)."
primer_bind 2187..2209
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 2206..2222
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2206..2222
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 2254..2556
/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="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind 2435..2454
/label=ccdB-fwd
/note="ccdB gene, forward primer"
primer_bind complement(2581..2598)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(2581..2597)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2590..2612)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(2806..2825)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2925..2947
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
ORIGIN
1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
2221 accatgatta cgaattcgcc aatacttgta tggcagttta aggtttacac ctataaaaga
2281 gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac gcccgggcga
2341 cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt
2401 tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt
2461 gtgccggtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc
2521 aaaaacgcca ttaacctgat gttctgggga atataaccat acaagtattg gaagcttggc
2581 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
2641 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
2701 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
2761 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
2821 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
2881 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
2941 tcagaggttt tcaccgtcat caccgaaacg cgcga
//
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