| 质粒名称: | pACBSR-hyg |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 87830 |
| 存储实验室: | Pep Charusanti |
|
载体骨架基本信息 |
|
| 载体名称: | pACBSR, which is based on pBAD |
| 载体出品公司: | -- |
| 载体抗性: | 潮霉素 |
| 空载体大小: | 6300 bp |
| 完整质粒大小: | 7300 bp |
| 载体修饰: | 1pACBSR中的camR替换为hygR 2。复制源是p15a ts |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 30 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | hyg |
| 物种来源: | -- |
| 插入大小: | 1040 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | bla |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BgI II（销毁后未知） |
| 3' 克隆位点: | XhoI（销毁后未知） |
| 5' 测序引物 | AGATCTCGGGGAAATGTGCGCGGAAC |
| 3' 测序引物 | CTCGAGGTATATATGAGTAAACTTGGTCTG |
|
文献引用方法 |
|
| 材料和方法部分: | pACBSR-hyg was a gift from Pep Charusanti (Addgene plasmid # 87830 ; http://n2t.net/addgene:87830 ; RRID:Addgene_87830) |
| 参考文献部分: | Capsule deletion via a lambda-Red knockout system perturbs biofilm formation and fimbriae expression in Klebsiella pneumoniae MGH 78578. Huang TW, Lam I, Chang HY, Tsai SF, Palsson BO, Charusanti P. BMC Res Notes. 2014 Jan 8;7:13. doi: 10.1186/1756-0500-7-13. 10.1186/1756-0500-7-13 PubMed 24398052 |
买家导航
pACBSR-hyg载体 pACBSRhyg质粒基本信息
pACBSR-hyg质粒图谱 pACBSRhyg载体图谱和pACBSR-hyg载体序列 pACBSRhyg质粒序列多克隆位点信息
pACBSR-hyg质粒 pACBSRhyg载体简介
pACBSR-hyg质粒序列 pACBSRhyg载体序列
LOCUS pACBSR-hyg 7660 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses lambda-red system with hyg selection marker.
ACCESSION .
VERSION .
KEYWORDS pACBSR-hyg
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7660)
AUTHORS Huang TW, Lam I, Chang HY, Tsai SF, Palsson BO, Charusanti P
TITLE Capsule deletion via a lambda-Red knockout system perturbs biofilm
formation and fimbriae expression in Klebsiella pneumoniae MGH
78578.
JOURNAL BMC Res Notes. 2014 Jan 8;7:13. doi: 10.1186/1756-0500-7-13.
PUBMED 24398052
REFERENCE 2 (bases 1 to 7660)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7660
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter complement(461..745)
/gene="araBAD"
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
primer_bind complement(519..538)
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
CDS 772..1650
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
primer_bind complement(1830..1849)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(1987..2009)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 2150..2169
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
CDS complement(2296..3321)
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
E"
promoter complement(3322..3426)
/gene="bla"
/label=AmpR promoter
primer_bind complement(3729..3746)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3863..4408)
/direction=LEFT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
"
primer_bind complement(3943..3962)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter complement(4658..4749)
/gene="bla"
/label=AmpR promoter
terminator 4768..4795
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 4887..4973
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 5126..5143
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 5203..5447
/label=-lambda tL3 terminator
/note="transcription terminator tL3 from phage lambda"
CDS complement(5448..6128)
/codon_start=1
/gene="exo"
/product="5' to 3' double-stranded DNA exonuclease in the
lambda Red system"
/label=Exo
/note="creates 3' single-stranded DNA tails"
/translation="MTPDIILQRTGIDVRAVEQGDDAWHKLRLGVITASEVHNVIAKPR
SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
EQWR"
CDS complement(6125..6910)
/codon_start=1
/gene="bet"
/product="single-stranded DNA binding recombinase in the
lambda Red system"
/label=Beta
/note="mediates pairing of a single-stranded DNA with a
complementary target"
/translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
CDS complement(6916..7332)
/codon_start=1
/gene="gam"
/product="inhibitor of the host RecBCD nuclease in the
lambda Red system"
/label=Gam
/note="protects the double-stranded DNA substrate for
recombination"
/translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY
IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR
AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV"
RBS 7341..7349
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
ORIGIN
1 gtatgccaga gacatcgggg tcaggtagtt ttcaaccagg ttgttcggga tggttttttt
61 gttgttaacg atgaacaggt tagccagttt gttgaaagct tggtgtttga aagtctgggc
121 gccccaggtg attaccaggt tacccaggtg gttaacacgt tcttttttgt gcggcgggga
181 cagtacccac tgatcgtaca gcagacatac gtggtccatg tatgctttgt ttttccactc
241 gaactgcata cagtaggttt taccttcatc acgagaacgg atgtaagcat cacccaggat
301 cagaccgata cctgcttcga actgttcgat gttcagttcg atcagctggg atttgtattc
361 tttcagcagt ttagagttcg gacccaggtt cattacctgg ttttttttga tgtttttcat
421 atgcatatag gtaccctcct gctagcccaa aaaaacgggt atggagaaac agtagagagt
481 tgcgataaaa agcgtcaggt aggatccgct aatcttatgg ataaaaatgc tatggcatag
541 caaagtgtga cgccgtgcaa ataatcaatg tggacttttc tgccgtgatt atagacactt
601 ttgttacgcg tttttgtcat ggctttggtc ccgctttgtt acagaatgct tttaataagc
661 ggggttaccg gtttggttag cgagaagagc cagtaaaaga cgcagtgacg gcaatgtctg
721 atgcaatatg gacaattggt ttcttctctg aatggcggga gtatgaaaag tatggctgaa
781 gcgcaaaatg atcccctgct gccgggatac tcgtttaatg cccatctggt ggcgggttta
841 acgccgattg aggccaacgg ttatctcgat ttttttatcg accgaccgct gggaatgaaa
901 ggttatattc tcaatctcac cattcgcggt cagggggtgg tgaaaaatca gggacgagaa
961 tttgtttgcc gaccgggtga tattttgctg ttcccgccag gagagattca tcactacggt
1021 cgtcatccgg aggctcgcga atggtatcac cagtgggttt actttcgtcc gcgcgcctac
1081 tggcatgaat ggcttaactg gccgtcaata tttgccaata cggggttctt tcgcccggat
1141 gaagcgcacc agccgcattt cagcgacctg tttgggcaaa tcattaacgc cgggcaaggg
1201 gaagggcgct attcggagct gctggcgata aatctgcttg agcaattgtt actgcggcgc
1261 atggaagcga ttaacgagtc gctccatcca ccgatggata atcgggtacg cgaggcttgt
1321 cagtacatca gcgatcacct ggcagacagc aattttgata tcgccagcgt cgcacagcat
1381 gtttgcttgt cgccgtcgcg tctgtcacat cttttccgcc agcagttagg gattagcgtc
1441 ttaagctggc gcgaggacca acgtatcagc caggcgaagc tgcttttgag caccacccgg
1501 atgcctatcg ccaccgtcgg tcgcaatgtt ggttttgacg atcaactcta tttctcgcgg
1561 gtatttaaaa aatgcaccgg ggccagcccg agcgagttcc gtgccggttg tgaagaaaaa
1621 gtgaatgatg tagccgtcaa gttgtcataa ttggtaacga atcagacaat tgacggcttg
1681 acggagtagc atagggtttg cagaatccct gcttcgtcca tttgacaggc acattatgca
1741 tcgatgataa gctgtcaaac atgagcagat cctctacgcc ggacgcatcg tggccggcat
1801 caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga
1861 tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc
1921 cgtggccggg ggactgttgg gcgccatctc cttctgcctc gcgcgtttcg gtgatgacgg
1981 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
2041 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
2101 catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag
2161 cagattgtac tgagagtgca cctgccattc atccgcttat tatcacttat tcaggcgtag
2221 caaccaggcg tttaagggca ccaataactg ccttaaaaaa actcgaggta tatatgagta
2281 aacttggtct gacagctatt cctttgccct cggacgagtg ctggggcgtc ggtttccact
2341 atcggcgagt acttctacac agccatcggt ccagacggcc gcgcttctgc gggcgatttg
2401 tgtacgcccg acagtcccgg ctccggatcg gacgattgcg tcgcatcgac cctgcgccca
2461 agctgcatca tcgaaattgc cgtcaaccaa gctctgatag agttggtcaa gaccaatgcg
2521 gagcatatac gcccggagcc gcggcgatcc tgcaagctcc ggatgcctcc gctcgaagta
2581 gcgcgtctgc tgctccatac aagccaacca cggcctccag aagaagatgt tggcgacctc
2641 gtattgggaa tccccgaaca tcgcctcgct ccagtcaatg accgctgtta tgcggccatt
2701 gtccgtcagg acattgttgg agccgaaatc cgcgtgcacg aggtgccgga cttcggggca
2761 gtcctcggcc caaagcatca gctcatcgag agcctgcgcg acggacgcac tgacggtgtc
2821 gtccatcaca gtttgccagt gatacacatg gggatcagca atcgcgcata tgaaatcacg
2881 ccatgtagtg tattgaccga ttccttgcgg tccgaatggg ccgaacccgc tcgtctggct
2941 aagatcggcc gcagcgatcg catccatggc ctccgcgacc ggctgcagaa cagcgggcag
3001 ttcggtttca ggcaggtctt gcaacgtgac accctgtgca cggcgggaga tgcaataggt
3061 caggctctcg ctgaattccc caatgtcaag cacttccgga atcgggagcg cggccgatgc
3121 aaagtgccga taaacataac gatctttgta gaaaccatcg gcgcagctat ttacccgcag
3181 gacatatcca cgccctccta catcgaagct gaaagcacga gattcttcgc cctccgagag
3241 ctgcatcagg tcggagacgc tgtcgaactt ttcgatcaga aacttctcga cagacgtcgc
3301 ggtgagttca ggctttttca tactcttcct ttttcaatat tattgaagca tttatcaggg
3361 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
3421 tccgcgcaca tttccccgag atctgcaaag tgcgtcgggt gatgctgcca acttactgat
3481 ttagtgtatg atggtgtttt tgaggtgctc cagtggcttc tgtttctatc agctgtccct
3541 cctgttcagc tactgacggg gtggtgcgta acggcaaaag caccgccgga catcagcgct
3601 agcggagtgt atactggctt actatgttgg cactgatgag ggtgtcagtg aagtgcttca
3661 tgtggcagga gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata caggatatat
3721 tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg
3781 gcttacgaac ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg
3841 agagggccgc ggcaaagccg tttttccata ggctccgccc ccctgacaag catcacgaaa
3901 tctgacgctc aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc
3961 cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc
4021 cgctgttatg gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt
4081 cgctccaagc tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc
4141 ggtaactatc gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc
4201 actggtaatt gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga
4261 aaggacaagt tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt
4321 agctcagaga accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga
4381 gattacgcgc agaccaaaac gatctcaaga agatcatctt attaatcaga taaaatattt
4441 ctagatttca gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata
4501 taagttgtaa ttctcatgtt tgacagctta tcatcggtgc acccaactga tcttcagcat
4561 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
4621 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
4681 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
4741 ataaacaaaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
4801 atttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
4861 gcaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
4921 caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcctg
4981 gcagttccct actctcgcat ggggagaccc cacactacca tcggcgctac ggcgtttcac
5041 ttctgagttc ggcatggggt caggtgggac caccgcgcta ctgccgccag gcaaattctg
5101 ttttatcaga ccgcttctgc gttctgattt aatctgtatc aggctgaaaa tcttctctca
5161 tccgccaaaa cagccaagct tgcatgcatt cttcgtctgt ttctactggt attggcacaa
5221 acctgattcc aatttgagca aggctatgtg ccatctcgat actcgttctt aactcaacag
5281 aagatgcttt gtgcatacag cccctcgttt attatttatc tcctcagcca gccgctgtgc
5341 tttcagtgga tttcggataa cagaaaggcc gggaaatacc cagcctcgct ttgtaacgga
5401 gtagacgaaa gtgattgcgc ctacccggat attatcgtga ggatgcgtca tcgccattgc
5461 tccccaaata caaaaccaat ttcagccagt gcctcgtcca ttttttcgat gaactccggc
5521 acgatctcgt caaaactcgc catgtacttt tcatcccgct caatcacgac ataatgcagg
5581 ccttcacgct tcatacgcgg gtcatagttg gcaaagtacc aggcattttt tcgcgtcacc
5641 cacatgctgt actgcacctg ggccatgtaa gctgacttta tggcctcgaa accaccgagc
5701 cggaacttca tgaaatcccg ggaggtaaac gggcatttca gttcaaggcc gttgccgtca
5761 ctgcataaac catcgggaga gcaggcggta cgcatacttt cgtcgcgata gatgatcggg
5821 gattcagtaa cattcacgcc ggaagtgaat tcaaacaggg ttctggcgtc gttctcgtac
5881 tgttttcccc aggccagtgc tttagcgtta acttccggag ccacaccggt gcaaacctca
5941 gcaagcaggg tgtggaagta ggacattttc atgtcaggcc acttctttcc ggagcggggt
6001 tttgctatca cgttgtgaac ttctgaagcg gtgatgacgc cgagccgtaa tttgtgccac
6061 gcatcatccc cctgttcgac agctctcaca tcgatcccgg tacgctgcag gataatgtcc
6121 ggtgtcatgc tgccaccttc tgctctgcgg ctttctgttt caggaatcca agagctttta
6181 ctgcttcggc ctgtgtcagt tctgacgatg cacgaatgtc gcggcgaaat atctgggaac
6241 agagcggcaa taagtcgtca tcccatgttt tatccagggc gatcagcaga gtgttaatct
6301 cctgcatggt ttcatcgtta accggagtga tgtcgcgttc cggctgacgt tctgcagtgt
6361 atgcagtatt ttcgacaatg cgctcggctt catccttgtc atagatacca gcaaatccga
6421 aggccagacg ggcacactga atcatggctt tatgacgtaa catccgtttg ggatgcgact
6481 gccacggccc cgtgatttct ctgccttcgc gagttttgaa tggttcgcgg cggcattcat
6541 ccatccattc ggtaacgcag atcggatgat tacggtcctt gcggtaaatc cggcatgtac
6601 aggattcatt gtcctgctca aagtccatgc catcaaactg ctggttttca ttgatgatgc
6661 gggaccagcc atcaacgccc accaccggaa cgatgccatt ctgcttatca ggaaaggcgt
6721 aaatttcttt cgtccacgga ttaaggccgt actggttggc aacgatcagt aatgcgatga
6781 actgcgcatc gctggcatca cctttaaatg ccgtctggcg aagagtggtg atcagttcct
6841 gtgggtcgac agaatccatg ccgacacgtt cagccagctt cccagccagc gttgcgagtg
6901 cagtactcat tcgttttata cctctgaatc aatatcaacc tggtggtgag caatggtttc
6961 aaccatgtac cggatgtgtt ctgccatgcg ctcctgaaac tcaacatcgt catcaaacgc
7021 acgggtaatg gattttttgc tggccccgtg gcgttgcaaa tgatcgatgc atagcgattc
7081 aaacaggtgc tggggcaggc ctttttccat gtcgtctgcc agttctgcct ctttctcttc
7141 acgggcgagc tgctggtagt gacgcgccca gctctgagcc tcaagacgat cctgaatgta
7201 ataagcgttc atggctgaac tcctgaaata gctgtgaaaa tatcgcccgc gaaatgccgg
7261 gctgattagg aaaacaggaa agggggttag tgaatgcttt tgcttgatct cagtttcagt
7321 attaatatcc attttttata acctccttag catgcctgca gacgtcgggc ccttatttca
7381 ggaaagtttc ggaggagata gtgttcggca gtttgtacat catctgcggg atcaggtacg
7441 gtttgatcag gttgtagaag atcaggtaag acatagaatc gatgtagatg atcggtttgt
7501 ttttgttgat ttttacgtaa cagttcagtt ggaatttgtt acgcagaccc ttaaccaggt
7561 attctacttc ttcgaaagtg aaagactggg tgttcagtac gatcgatttg ttggtagagt
7621 ttttgttgta atcccattta ccaccatcat ccatgaacca
//
