pPTRII

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联系方式:I47-825O-882O

相关技术服务:质粒构建    基因合成    质粒大提

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pPTR II载体基本信息

出品公司: Takara
载体名称: pPTRII
质粒类型: 曲霉表达质粒,大肠杆菌-曲霉穿梭质粒,曲霉表达载体
高拷贝/低拷贝: 高拷贝
克隆方法: 限制性内切酶,多克隆位点
启动子: Lac
载体大小: 10031 bp
5’测序引物: M13-R: CAGGAAACAGCTATGAC
3’测序引物: M13-F: TGTAAAACGACGGCCAGT
载体抗性: 氨苄青霉素
克隆菌株: DH5 alpha
宿主细胞(系): 曲霉菌
产品目录号: 3622
稳定性: 瞬表达
组成型/诱导型: 组成型
病毒/非病毒: 非病毒
相关载体: pPTRI
载体说明书: pPTRII载体说明书
备注: pPTRII载体是曲霉表达载体,能够在曲霉菌中进行自我复制。pPTRII载体可以用于多种曲霉菌(Aspergillus species),例如A. oryzae, A . niger, A. terreus and A . fumigatus。pPTRI载体在曲霉菌中的筛选抗性是Pyrithiamine,中文名是抗硫胺素, 也叫吡啶硫铵。 本载体在构建质粒时,可以利用蓝白斑筛选进行阳性插入克隆验证。

pPTR II载体质粒图谱和多克隆位点信息

pPTRII载体图谱


pPTRII多克隆位点

pPTR II载体简介

pPTR II是一可以在曲霉菌中可以自主复制的表达载体质粒。该载体具有氨苄西林抗性基因 Ampr作为大肠杆菌筛选抗性。
 
在曲霉中,筛选基因是抗硫胺素(Pyrithiamine) ,抗性基因为ptr A基因。本质粒具有多克隆位点,包含 Hind III,Pst I,Sma I和Kpn I。
 
在载体lac Z区域,任何DNA插入到这个区域 使用含有IPTG和X-Gal的平板可以很容易地进行蓝白斑筛选阳性克隆载体。

pPTR II载体序列

LOCUS       pTR II Vector              10031 bp DNA     circular SYN 23-JUL-2021
COMMENT	    Created by Takara Bio Inc. http://www.biofeng.com
COMMENT     Cat. No. 3622
FEATURES             Location/Qualifiers
     gene            complement(2943..4950)
                     /label="ptrA"
                     /note="pyrithiamine resistance gene useful as a dominant selectable marker in
                     Aspergillus"
     CDS             4968..5828
                     /label="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     rep_origin      5999..6587
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        6911..6941
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    6949..6965
                     /label="lac operator"
                     /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     6973..6989
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             6985..7311
                     /label="lacZ-alpha"
     misc_feature    7002..7058
                     /label="MCS"
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(7059..7075)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      join(7416..10031,1..2633)
                     /label="AMA1"
                     /note="replication origin from Aspergillus nidulans
                     (Aleksenko and Clutterbuck, 1996)"
ORIGIN
        1 ctgcagattt tgacctagtt aacaaggcac atcatgctat cacttccaac actcttgctg
       61 agcgcgttct gccttgcctt tctggttcta cgaagtatcc agtcccgcct gcaacgccgc
      121 agaaacgcca aaaggtgggg ttgtcaaccg ccagctagag ctccatcggg attctttggc
      181 attccgccgt tcctcggtgg tcgcagataa ctccctaaat tccctgagct aactctaagt
      241 cgaccatgcc gtttatggtt agcgcctccc aaaaaggaat ggccgactta gagttacctc
      301 ttgaccgact ttttctttcc tcccccttac atttcgttac cacaacacat tcctatatcc
      361 aaactccagg tacataacta gtcgaaatct cttttaaatc tagtcaagaa ctagatactt
      421 aaccttcata ctgcttagag atagcctttg aagctcttat ttcgctcttt gtctcactct
      481 caccttcctc ctcctcctcc aatccttttt gctcgggact agtccaagac tagtcaacga
      541 ttagtatgcc gagcattcgc gataacgatc tccgaaagtc cccagagtac tgtcactatc
      601 tcgaggcagt taaggacggg gagcttacgc tgccggattt caagatagta agccgacccg
      661 actagcttta agcctagtta ctgactagtc cacaggacga caatggcgtg cctgatatcc
      721 atccatatga agtctactgc cgagtgaagg gatgcctcaa gcgtacagtg agtctactgc
      781 tatactagtt tctgactagt tattaactag ttgacaggtt ccctctgcca acagaaacat
      841 attggtcaag cacttgaagg acaagaactc ccacggcatg gagtttacat tgcacaatgg
      901 acctcccact atgaaggaac tgatggaggc caaaggcaag tcctatctag attacttggt
      961 gactagtccc tgactagtca agtagcatgg tatgaaggct tgtttgaagg cactgttctc
     1021 ccaaccccga ctcctaccaa gaagcgcaag cgagctgcgt aagtttctgt gagtctaact
     1081 agtgtattag ctaatatata gcagcaccaa gtccaaggac cacaatacta agggtgtcga
     1141 gaattcgtga gtttcttctc ccatttcaac tagtccttga ctagtcacta actactttgc
     1201 agcaacgagg gtgaagctgg aaatgatcag gacaatggcg agggcccgta agtacagcca
     1261 ttcaatgcag actagttgct aactaatctg tgactagttc aagtggtccg tacgccgtgc
     1321 atacccctgt gactggtagg aatttgagca agcctgtctt gccgcgcgat gagaaaggaa
     1381 aggcaagtta cattcagccc cgtacctagg atcagtgcta atttataacc tctagccact
     1441 ctttatgcag atccgccgtg agggtagcaa ggcagctaaa tcagctggtg agaaaggaac
     1501 cataccctgc aagacctgtc gcaacgcaaa gggcaaaggt aagctatcca agctagtttg
     1561 ggactagatt ctaactagtc tcagcaccgt gtggttcaaa gccatattgc gagttttggc
     1621 gctttttctc atcgattgac gaggcaaagg gagcgagtat gcaacctcgt aagtcagact
     1681 cagacaaaga ccaactagtt attgaccagt cttctagaag gctctgttgt ggatcttgag
     1741 gccctggaga gttcctccaa caatccggag acaagcaagt cctcgtcgga ctagtcacta
     1801 actagactct aactagttgc agacatggat aatgcaaaag agacaagcaa tgaagaaagt
     1861 ggtaagacat ttctcctttg tggttctgga ctagtctttg actagtcaca gtcttaaaca
     1921 aggaaaatga gcatgaaaat gaggaggaaa aggctgctga gcccgaggaa gtgcagggtg
     1981 atggcagaca tggtaggtta ataccttgtt agttattgct agtcactgac tagtcaataa
     2041 ctagtctctg aacaccttgc aatcactccg tttgcgcagc tgaacagtgg tgaggataat
     2101 agtagtaagt tattctagct tcagagttat aggagactag atactaacta gtattagttg
     2161 caactaacct ggatctcaga gactttggcc tcaatctaga atctatctag ttgtcaacta
     2221 gactgtggta tcattgtctt ttattttcct agtcctggaa ctagcttcta actagtctcc
     2281 ctaatatgtg gctgtcttgt tttttttttt tgtttcccta cccggatatc tagtcccctt
     2341 ctaggttctg ttaacctctc gggctctgat ttagtttaac gcaaacctga gattagtttc
     2401 taactagtct ctaggttttc tatccacctt taattgtaat aataaataca agcaacgttt
     2461 atacgtcaaa agcatttata aacttttacc ctaaagtagc ttgcttgtgt gtttagttta
     2521 taattagtct cttattaatt tgatgtaggt aagcccgcca caaatatata tttttaacaa
     2581 gataccgtgg aaaaacttcg tgctatcaca aaacagtata caaaaaataa gctggctttc
     2641 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2701 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     2761 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2821 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     2881 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     2941 aaatggggtg acgatgagcc gctcttgcat ctttgtttgt attatactgt ctttcttgtt
     3001 acacataatt attctagaat gccccaccgt tacatacggg acacagccat ttacatatgc
     3061 atgtggatta cgagctaacg agttcattca aatctcagaa ctatcacata atcatcattc
     3121 ccctatcgtc aaagaccgta agacaaatcc ggttcatgca ctgaacccat tcgggtagtg
     3181 agtcatttac tcagcacact cgcgctgacg ctcgtcgaac accttcaatg cctcctcggc
     3241 agccttgaca ccactgagaa ccatggcacc gaaggtaggg cccatgcggt taaagccatc
     3301 aatttcagac agctccatac cgccgattat caagccctta gtaacctcgc gggtgttctt
     3361 gacgatggca tcctcggccg agctcatgtc gagaccacgc atgccaccta gcttgtcgac
     3421 gctgcccatg gacaccaagc gcttcgcaca gaaggcgccg aatggcccat cgtgaccagt
     3481 ggtactgatg atgacaggag cgttgatagt gttggggtcc atgcaggagt gatcatcgtg
     3541 gtgaagggtg accagcgtcc agttgacgac aacaccagca atctgggggt tgccgttctc
     3601 ggtcggacgg gtgatcaagt cctcaacagc ggtagcattg aagagcttga cattggggaa
     3661 ggagagaacc ttcgacatga gtgtcgaggt aaacagggag gcgtgcttga cgacaacgta
     3721 gttggggttt gcgtcctctt cgtaaggaac acccagctcg ttcaggaaga cttccgcggg
     3781 acggcgcatg accatagcag aaaagagttg gccacccaac caggcaccgc cacctattgc
     3841 acgttagttc cggaaagctg agtgcaaggc aatccatcat ggactactga ccaggagaga
     3901 cgctggcctc gacgatagca atcttcaggt ccggacgagc cttggccaag acgtacgcag
     3961 tgctcagacc gcaggaacca gcaccaacaa tgacaacgtc actttcagcg tacttgtcca
     4021 ggtcctcaaa gtaacgtctg gtcatggcac gagagacctg gctttcgcgg ataggggcga
     4081 acttgaactc gtcccacttg ccaccgaaat ggtccaacag cttggtctga gaagctccct
     4141 caacggggac ggtctcagaa accacgacct tacccttgag gccggtagcg gccacagtgg
     4201 gttcgtagat ggcagctgga ggagacatgt ttcaagttgc aatgactatc atctgttagc
     4261 cattccatca acaggaagaa cgagagaagg catgaccctt ttcgctggta ttatccagat
     4321 caagttttag ccgtataatc tcagaacgaa cccagtccat cgatgccatg tccttctaga
     4381 ctaggatcct agagtctagg gcccagctta gggagggcat gtgaatgcat cgatgactgg
     4441 gaacgaacac cggcccacgc caaagacgtt acctaagata ccttgatcat tgtgagagtc
     4501 cagccaaaag tattccatga cttccatcgt atgccctcta gagggctaat cgaggagtgt
     4561 atttacattg tcggttggtt tgggaactat agaagatggt cagttattcc aatcaccaaa
     4621 ggtttatcga agggaggaag acttgttcag tttcgtccga ggacttttgg aattcaaatc
     4681 tgagatagag aattgtgtgg gatgagagga aaaggaagaa gatggtgggg ttcagaagga
     4741 ggggttgagt taaatagcat gggttgagtc aacgtgataa gggcactata ccgtatagat
     4801 cagcggcacc cgattctatc cgttcctttt gctcctcttt agctttgacc ggtgagccgg
     4861 acaagaaaca agtgaaatca tcctgacatc ggcggacgat ctcctagctt ttacatttcg
     4921 ttaccaatgg gatcccgtaa tcaattgccc attgaaaaag gaagagtatg agtattcaac
     4981 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     5041 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     5101 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     5161 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5221 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     5281 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5341 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     5401 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5461 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5521 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     5581 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5641 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5701 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5761 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5821 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5881 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5941 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6001 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6061 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6121 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     6181 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6241 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6301 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6361 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6421 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6481 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6541 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6601 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6661 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     6721 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     6781 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     6841 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     6901 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     6961 aacaatttca cacaggaaac agctatgacc atgattacgc caagcttgca tgcctgcagg
     7021 tcgactctag aggatccccg ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt
     7081 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     7141 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     7201 ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     7261 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     7321 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     7381 tcgctttctt cccttccttt ctcgccacgt tcgccagctt attttttgta tactgttttg
     7441 tgatagcacg aagtttttcc acggtatctt gttaaaaata tatatttgtg gcgggcttac
     7501 ctacatcaaa ttaataagag actaattata aactaaacac acaagcaagc tactttaggg
     7561 taaaagttta taaatgcttt tgacgtataa acgttgcttg tatttattat tacaattaaa
     7621 ggtggataga aaacctagag actagttaga aactaatctc aggtttgcgt taaactaaat
     7681 cagagcccga gaggttaaca gaacctagaa ggggactaga tatccgggta gggaaacaaa
     7741 aaaaaaaaac aagacagcca catattaggg agactagtta gaagctagtt ccaggactag
     7801 gaaaataaaa gacaatgata ccacagtcta gttgacaact agatagattc tagattgagg
     7861 ccaaagtctc tgagatccag gttagttgca actaatacta gttagtatct agtctcctat
     7921 aactctgaag ctagaataac ttactactat tatcctcacc actgttcagc tgcgcaaacg
     7981 gagtgattgc aaggtgttca gagactagtt attgactagt cagtgactag caataactaa
     8041 caaggtatta acctaccatg tctgccatca ccctgcactt cctcgggctc agcagccttt
     8101 tcctcctcat tttcatgctc attttccttg tttaagactg tgactagtca aagactagtc
     8161 cagaaccaca aaggagaaat gtcttaccac tttcttcatt gcttgtctct tttgcattat
     8221 ccatgtctgc aactagttag agtctagtta gtgactagtc cgacgaggac ttgcttgtct
     8281 ccggattgtt ggaggaactc tccagggcct caagatccac aacagagcct tctagaagac
     8341 tggtcaataa ctagttggtc tttgtctgag tctgacttac gaggttgcat actcgctccc
     8401 tttgcctcgt caatcgatga gaaaaagcgc caaaactcgc aatatggctt tgaaccacac
     8461 ggtgctgaga ctagttagaa tctagtccca aactagcttg gatagcttac ctttgccctt
     8521 tgcgttgcga caggtcttgc agggtatggt tcctttctca ccagctgatt tagctgcctt
     8581 gctaccctca cggcggatct gcataaagag tggctagagg ttataaatta gcactgatcc
     8641 taggtacggg gctgaatgta acttgccttt cctttctcat cgcgcggcaa gacaggcttg
     8701 ctcaaattcc taccagtcac aggggtatgc acggcgtacg gaccacttga actagtcaca
     8761 gattagttag caactagtct gcattgaatg gctgtactta cgggccctcg ccattgtcct
     8821 gatcatttcc agcttcaccc tcgttgctgc aaagtagtta gtgactagtc aaggactagt
     8881 tgaaatggga gaagaaactc acgaattctc gacaccctta gtattgtggt ccttggactt
     8941 ggtgctgcta tatattagct aatacactag ttagactcac agaaacttac gcagctcgct
     9001 tgcgcttctt ggtaggagtc ggggttggga gaacagtgcc ttcaaacaag ccttcatacc
     9061 atgctacttg actagtcagg gactagtcac caagtaatct agataggact tgcctttggc
     9121 ctccatcagt tccttcatag tgggaggtcc attgtgcaat gtaaactcca tgccgtggga
     9181 gttcttgtcc ttcaagtgct tgaccaatat gtttctgttg gcagagggaa cctgtcaact
     9241 agttaataac tagtcagaaa ctagtatagc agtagactca ctgtacgctt gaggcatccc
     9301 ttcactcggc agtagacttc atatggatgg atatcaggca cgccattgtc gtcctgtgga
     9361 ctagtcagta actaggctta aagctagtcg ggtcggctta ctatcttgaa atccggcagc
     9421 gtaagctccc cgtccttaac tgcctcgaga tagtgacagt actctgggga ctttcggaga
     9481 tcgttatcgc gaatgctcgg catactaatc gttgactagt cttggactag tcccgagcaa
     9541 aaaggattgg aggaggagga ggaaggtgag agtgagacaa agagcgaaat aagagcttca
     9601 aaggctatct ctaagcagta tgaaggttaa gtatctagtt cttgactaga tttaaaagag
     9661 atttcgacta gttatgtacc tggagtttgg atataggaat gtgttgtggt aacgaaatgt
     9721 aagggggagg aaagaaaaag tcggtcaaga ggtaactcta agtcggccat tcctttttgg
     9781 gaggcgctaa ccataaacgg catggtcgac ttagagttag ctcagggaat ttagggagtt
     9841 atctgcgacc actggactct caggctttcc attggggtac taacatagcc atcaaatgcc
     9901 ggcccgtgtt taatctcctc gagatggctg ggctcacatg cgttctactc aaaaactctt
     9961 atacctaggc cctctcccaa tcgtgtatgt ttggaccttc ctctctcaca ctttggataa
    10021 catcaagcat a
//

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