| 质粒名称: | pCACTUS |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 167559 |
| 存储实验室: | Renato Morona |
|
载体骨架基本信息 |
|
| 载体名称: | pCACTUS |
| 载体出品公司: | Clontech |
| 空载体大小: | 6201bp |
| 完整质粒大小: | 6201bp |
| 载体类型: | Suicide cloning vector, temperature sensitive. |
| 载体抗性: | Chloramphenicol, 25 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: | pCACTUS was a gift from Renato Morona (Addgene plasmid # 167559 ; http://n2t.net/addgene:167559 ; RRID:Addgene_167559) |
| 参考文献部分: | Molecular, genetic, and topological characterization of O-antigen chain length regulation in Shigella flexneri. Morona R, van den Bosch L, Manning PA. J Bacteriol. 1995 Feb;177(4):1059-68. doi: 10.1128/jb.177.4.1059-1068.1995. 10.1128/jb.177.4.1059-1068.1995 PubMed 7532168 |
买家导航
pCACTUS载体质粒基本信息
pCACTUS质粒图谱载体图谱和pCACTUS载体序列质粒序列多克隆位点信息
pCACTUS质粒载体简介
pCACTUS质粒序列载体序列
LOCUS pCACTUS 6202 bp DNA circular SYN 25-AUG-2024
DEFINITION Suicide vector with the sacB gene (as a contraselective marker),
CmlR gene, and temperature sensitive ori, used to perform allelic
exchange mutagenesis..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6202)
AUTHORS Morona R, van den Bosch L, Manning PA
TITLE Molecular, genetic, and topological characterization of O-antigen
chain length regulation in Shigella flexneri.
JOURNAL J Bacteriol. 1995 Feb;177(4):1059-68. doi:
10.1128/jb.177.4.1059-1068.1995.
PUBMED 7532168
REFERENCE 2 (bases 1 to 6202)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1128/jb.177.4.1059-1068"; journalName: "J Bacteriol"; date:
"1995-02"; volume: "177"; issue: "4"; pages: "1059-68"
FEATURES Location/Qualifiers
source 1..6202
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 545..767
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
CDS 815..1765
/codon_start=1
/gene="rep101 encoding an A56V mutation"
/product="temperature-sensitive version of the RepA protein
needed for replication with the pSC101 origin (Armstrong et
al., 1984)"
/label=Rep101(Ts)
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
promoter 2799..2901
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 2902..3561
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(2968..2987)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter 3791..4236
/gene="sacR"
/label=sacB promoter
/note="sacB promoter and control region"
CDS 4237..5658
/codon_start=1
/gene="Bacillus subtilis sacB"
/product="secreted levansucrase that renders bacterial
growth sensitive to sucrose"
/label=SacB
/note="negative selection marker"
/translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
VKDSILEQGQLTVNK"
misc_feature 6066..6122
/label=MCS
/note="pUC18/19 multiple cloning site"
ORIGIN
1 ggcctcaacc tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga
61 tccccgacag taagacgggt aagcctgttg atgataccgc tgccttactg ggtgcattag
121 ccagtctgaa tgacctgtca cgggataatc cgaagtggtc agactggaaa atcagagggc
181 aggaactgct gaacagcaaa aagtcagata gcaccacata gcagacccgc cataaaacgc
241 cctgagaagc ccgtgacggg cttttcttgt attatgggta gtttccttgc atgaatccat
301 aaaaggcgcc tgtagtgcca tttaccccca ttcactgcca gagccgtgag cgcagcgaac
361 tgaatgtcac gaaaaagaca gcgactcagg tgcctgatgg tcggagacaa aaggaatatt
421 cagcgatttg cccgagcttg cgagggtgct acttaagcct ttagggtttt aaggtctgtt
481 ttgtagagga gcaaacagcg tttgcgacat ccttttgtaa tactgcggaa ctgactaaag
541 tagtgagtta tacacagggc tgggatctat tctttttatc tttttttatt ctttctttat
601 tctataaatt ataaccactt gaatataaac aaaaaaaaca cacaaaggtc tagcggaatt
661 tacagagggt ctagcagaat ttacaagttt tccagcaaag gtctagcaga atttacagat
721 acccacaact caaaggaaaa ggactagtaa ttatcattga ctagcccatc tcaattggta
781 tagtgattaa aatcacctag accaattgag atgtatgtct gaattagttg ttttcaaagc
841 aaatgaacta gcgattagtc gctatgactt aacggagcat gaaaccaagc taattttatg
901 ctgtgtggca ctactcaacc ccacgattga aaaccctaca aggaaagaac ggacggtatc
961 gttcacttat aaccaatacg ttcagatgat gaacatcagt agggaaaatg cttatggtgt
1021 attagctaaa gcaaccagag agctgatgac gagaactgtg gaaatcagga atcctttggt
1081 taaaggcttt gagattttcc agtggacaaa ctatgccaag ttctcaagcg aaaaattaga
1141 attagttttt agtgaagaga tattgcctta tcttttccag ttaaaaaaat tcataaaata
1201 taatctggaa catgttaagt cttttgaaaa caaatactct atgaggattt atgagtggtt
1261 attaaaagaa ctaacacaaa agaaaactca caaggcaaat atagagatta gccttgatga
1321 atttaagttc atgttaatgc ttgaaaataa ctaccatgag tttaaaaggc ttaaccaatg
1381 ggttttgaaa ccaataagta aagatttaaa cacttacagc aatatgaaat tggtggttga
1441 taagcgaggc cgcccgactg atacgttgat tttccaagtt gaactagata gacaaatgga
1501 tctcgtaacc gaacttgaga acaaccagat aaaaatgaat ggtgacaaaa taccaacaac
1561 cattacatca gattcctacc tacataacgg actaagaaaa acactacacg atgctttaac
1621 tgcaaaaatt cagctcacca gttttgaggc aaaatttttg agtgacatgc aaagtaagta
1681 tgatctcaat ggttcgttct catggctcac gcaaaaacaa cgaaccacac tagagaacat
1741 actggctaaa tacggaagga tctgaggttc ttatggctct tgtatctatc agtgaagcat
1801 caagactaac aaacaaaagt agaacaactg ttcaccgtta catatcaaag ggaaaactgt
1861 ccatatgcac agatgaaaac ggtgtaaaaa agatagatac atcagagctt ttacgagttt
1921 ttggtgcatt taaagctgtt caccatgaac agatcgacaa tgtaacagat gaacagcatg
1981 taacacctaa tagaacaggt gaaaccagta aaacaaagca actagaacat gaaattgaac
2041 acctgagaca acttgttaca gctcaacagt cacacataga cagcctgaaa caggcgatgc
2101 tgcttatcga atcaaagctg ccgacaacac gggagccagt gacgcctccc gtggggaaaa
2161 aatcatggca attctggaag aaatagcgct ttcagccggc aaaccggctg aagccggatc
2221 tgcgattctg ataacaaact agcaacacca gaacagcccg tttgcgggca gcaaaacccg
2281 tacttttgga cgttccggcg gttttttgtg gcgagtggtg ttcgggcggt gcgcgcaaga
2341 tccattatgt taaacgggcg agtttacatc tcaaaaccgc ccgcttaaca ccatcagaaa
2401 tcctcagcgc gattttaagc accaaccccc ccccgtaaca cccaaatcca tactgaaagt
2461 ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg catcctcccg
2521 acaacacaga ccattccgtg gcaaagcaaa agttcagaat caccaactgg tccacctaca
2581 acaaagctct catcaaccgt ggctccctca ctttctggct ggatgatgag gcgattcagg
2641 cctggtatga gtcggcaaca ccttcatcac gaggaaggcc gccctttcgt cttcgaataa
2701 atacctgtga cggaagatca cttcgcagaa taaataaatc ctggtgtccc tgttgatacc
2761 gggaagccct gggccaactt ttggcgaaaa tgagacgttg atcggcacgt aagaggttcc
2821 aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt
2881 tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata
2941 tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat
3001 aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac
3061 aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattc
3121 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
3181 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
3241 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
3301 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc
3361 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
3421 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
3481 gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
3541 gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggt
3601 tgctacgcct gaataagtga taataagcgg atgaatggca gaaattcgaa agcaaattcg
3661 acccggtcgt cggttcaggg cagggtcgtt aaatagccgc ttatgtctat tgctggttta
3721 ccggtttatt gactaccgga agcagtgtga ccgtgtgctt ctcaaatgcc tgagatcctt
3781 tttaacccat cacatatacc tgccgttcac tattatttag tgaaatgaga tattatgata
3841 ttttctgaat tgtgattaaa aaggcaactt tatgcccatg caacagaaac tataaaaaat
3901 acagagaatg aaaagaaaca gatagatttt ttagttcttt aggcccgtag tctgcaaatc
3961 cttttatgat tttctatcaa acaaaagagg aaaatagacc agttgcaatc caaacgagag
4021 tctaatagaa tgaggtcgaa aagtaaatcg cgcgggtttg ttactgataa agcaggcaag
4081 acctaaaatg tgtaaagggc aaagtgtata ctttggcgtc accccttaca tattttaggt
4141 ctttttttat tgtgcgtaac taacttgcca tcttcaaaca ggagggctgg aagaagcaga
4201 ccgctaacac agtacataaa aaaggagaca tgaacgatga acatcaaaaa gtttgcaaaa
4261 caagcaacag tattaacctt tactaccgca ctgctggcag gaggcgcaac tcaagcgttt
4321 gcgaaagaaa cgaaccaaaa gccatataag gaaacatacg gcatttccca tattacacgc
4381 catgatatgc tgcaaatccc tgaacagcaa aaaaatgaaa aatatcaagt tcctgaattc
4441 gattcgtcca caattaaaaa tatctcttct gcaaaaggcc tggacgtttg ggacagctgg
4501 ccattacaaa acgctgacgg cactgtcgca aactatcacg gctaccacat cgtctttgca
4561 ttagccggag atcctaaaaa tgcggatgac acatcgattt acatgttcta tcaaaaagtc
4621 ggcgaaactt ctattgacag ctggaaaaac gctggccgcg tctttaaaga cagcgacaaa
4681 ttcgatgcaa atgattctat cctaaaagac caaacacaag aatggtcagg ttcagccaca
4741 tttacatctg acggaaaaat ccgtttattc tacactgatt tctccggtaa acattacggc
4801 aaacaaacac tgacaactgc acaagttaac gtatcagcat cagacagctc tttgaacatc
4861 aacggtgtag aggattataa atcaatcttt gacggtgacg gaaaaacgta tcaaaatgta
4921 cagcagttca tcgatgaagg caactacagc tcaggcgaca accatacgct gagagatcct
4981 cactacgtag aagataaagg ccacaaatac ttagtatttg aagcaaacac tggaactgaa
5041 gatggctacc aaggcgaaga atctttattt aacaaagcat actatggcaa aagcacatca
5101 ttcttccgtc aagaaagtca aaaacttctg caaagcgata aaaaacgcac ggctgagtta
5161 gcaaacggcg ctctcggtat gattgagcta aacgatgatt acacactgaa aaaagtgatg
5221 aaaccgctga ttgcatctaa cacagtaaca gatgaaattg aacgcgcgaa cgtctttaaa
5281 atgaacggca aatggtacct gttcactgac tcccgcggat caaaaatgac gattgacggc
5341 attacgtcta acgatattta catgcttggt tatgtttcta attctttaac tggcccatac
5401 aagccgctga acaaaactgg ccttgtgtta aaaatggatc ttgatcctaa cgatgtaacc
5461 tttacttact cacacttcgc tgtacctcaa gcgaaaggaa acaatgtcgt gattacaagc
5521 tatatgacaa acagaggatt ctacgcagac aaacaatcaa cgtttgcgcc aagcttcctg
5581 ctgaacatca aaggcaagaa aacatctgtt gtcaaagaca gcatccttga acaaggacaa
5641 ttaacagtta acaaataaaa acgcaaaaga aaatgccgat tgaggccagt ttgctcaggc
5701 tctccccgtg gaggtaataa ttgacgatat gatcatttat tctgcctccc agagcctgat
5761 aaaaacggtt agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
5821 cctgcgatgc agatccggaa cataatggtg cagggcgctt gtttcggcgt gggtatggtg
5881 gcaggccgcc ccgtggccgg gggactgttg ggcgctgccg gcacctgtcc tacgagttgc
5941 atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag
6001 gagctaccgg cagcggtgcg gactgttgta actcagaata agaaatgagg ccgctcatgg
6061 cgttggaatt cgagctcggt acccggggat cctctagagt cgacctgcag gcatgcaagc
6121 ttgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tggcaccatt
6181 ccttgcggcg gcggtgctca ac
//
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