| 质粒名称: | pDO23A |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 27367 |
| 存储实验室: | Dirk Schnappinger |
|
载体骨架基本信息 |
|
| 载体名称: | pDONRP2R-P3 |
| 载体出品公司: | Gateway(R) (Invitrogen) |
| 空载体大小: | 3012 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达；网关供体载体 |
| 筛选标记: | -- |
| 载体抗性: | 氯霉素和氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DB3.1 |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | pUC/ori, bla,ori, AmpR |
| 物种来源: | -- |
| 插入大小: | 1813 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gateway克隆 |
| 5’克隆位点: | PciI（如果已销毁，则未知） |
| 3’克隆位点: | ZraI (Insert), EcoRV (Vector)（如果已销毁，则未知） |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pDO23A was a gift from Dirk Schnappinger (Addgene plasmid # 27367 ; http://n2t.net/addgene:27367 ; RRID:Addgene_27367) |
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 pDO23A 4825 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pDO23A
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4825)
TITLE Blumenthal A.
REFERENCE 2 (bases 1 to 4825)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4825
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(61..649)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(141..160)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(820..1680)
/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 1443..1462
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1681..1785)
/gene="bla"
/label=AmpR promoter
protein_bind 1847..2078
/gene="mutant version of attP"
/label=attP3
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS complement(2435..2740)
/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(2537..2556)
/label=ccdB-fwd
/note="ccdB gene, forward primer"
CDS complement(3082..3741)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 3656..3675
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(3742..3844)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
protein_bind 4002..4233
/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(4271..4288)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4271..4287)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4280..4302)
/label=M13/pUC Forward
/note="In lacZ gene"
terminator 4351..4437
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 4529..4556
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
primer_bind complement(4715..4732)
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
61 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
121 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
181 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
241 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
301 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
361 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
421 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
481 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
541 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
601 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
661 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
721 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
781 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
841 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
901 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
961 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
1021 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
1081 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
1141 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
1201 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
1261 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
1321 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
1381 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
1441 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
1501 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
1561 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
1621 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
1681 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
1741 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
1801 agtgccacct gacatcgcgg ccgcgttaac gctaccatgg agctccaaat aatgatttta
1861 ttttgactga tagtgacctg ttcgttgcaa caaattgata agcaatgctt ttttataatg
1921 ccaactttgt ataataaagt tgaacgagaa acgtaaaatg atataaatat caatatatta
1981 aattagattt tgcataaaaa acagactaca taatactgta aaacacaaca tatgcagtca
2041 ctatgaatca actacttaga tggtattagt gacctgtaga attcgagctc gaccgacagc
2101 cttccaaatg ttcttctcaa acggaatcgt cgtatccagc ctactcgcta ttgtcctcaa
2161 tgccgtatta aatcataaaa agaaataaga aaaagaggtg cgagcctctt ttttgtgtga
2221 caaaataaaa acatctacct attcatatac gctagtgtca tagtcctgaa aatcatctgc
2281 atcaagaaca atttcacaac tcttatactt ttctcttaca agtcgttcgg cttcatctgg
2341 attttcagcc tctatactta ctaaacgtga taaagtttct gtaatttcta ctgtatcgac
2401 ctgcagactg gctgtgtata agggagcctg acatttatat tccccagaac atcaggttaa
2461 tggcgttttt gatgtcattt tcgcggtggc tgagatcagc cacttcttcc ccgataacgg
2521 agaccggcac actggccata tcggtggtca tcatgcgcca gctttcatcc ccgatatgca
2581 ccaccgggta aagttcacgg gagactttat ctgacagcag acgtgcactg gccaggggga
2641 tcaccatccg tcgcccgggc gtgtcaataa tatcactctg tacatccaca aacagacgat
2701 aacggctctc tcttttatag gtgtaaacct taaactgcat ttcaccagtc cctgttctcg
2761 tcagcaaaag agccgttcat ttcaataaac cgggcgacct cagccatccc ttcctgattt
2821 tccgctttcc agcgttcggc acgcagacga cgggcttcat tctgcatggt tgtgcttacc
2881 agaccggaga tattgacatc atatatgcct tgagcaactg atagctgtcg ctgtcaactg
2941 tcactgtaat acgctgcttc atagcacacc tctttttgac atacttcggg tatacatatc
3001 agtatatatt cttataccgc aaaaatcagc gcgcaaatac gcatactgtt atctggcttt
3061 tagtaagccg gatccacgcg attacgcccc gccctgccac tcatcgcagt actgttgtaa
3121 ttcattaagc attctgccga catggaagcc atcacagacg gcatgatgaa cctgaatcgc
3181 cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc
3241 gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt
3301 ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc
3361 gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc
3421 actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac
3481 actatcccat atcaccagct caccgtcttt cattgccata cggaattccg gatgagcatt
3541 catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac
3601 ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac
3661 tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata
3721 tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
3781 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg
3841 atcaacgtct cattttcgcc aaaagttggc ccagggcttc ccggtatcaa cagggacacc
3901 aggatttatt tattctgcga agtgatcttc cgtcacaggt atttattcgg cgcaaagtgc
3961 gtcgggtgat gctgccaact tagtcgacgc tctagagctc caaataatga ttttattttg
4021 actgatagtg acctgttcgt tgcaacaaat tgataagcaa tgctttttta taatgccaac
4081 tttgtacaag aaagttgaac gagaaacgta aaatgatata aatatcaata tattaaatta
4141 gattttgcat aaaaaacaga ctacataata ctgtaaaaca caacatatgc agtcactatg
4201 aaccaactac ttagatggta ttagtgacct gtagaattcg agctctagag ctgcagggcc
4261 cgagcttaag actggccgtc gttttacaac gtcgtgactg ggaaaacatc catgctagcg
4321 ttaacgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcggaagac
4381 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
4441 ccgggagcgg atttgaacgt tgtgaagcaa cggcccggag ggtggcgggc aggacgcccg
4501 ccataaactg ccaggcatca aactaagcag aaggccatcc tgacggatgg cctttttgcg
4561 tttctacaaa ctcttcctgg ctagcggtac gcgtattaat tgcgttgcgc tcactgcccg
4621 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
4681 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
4741 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
4801 aatcagggga taacgcagga aagaa
//
