P1093 pCT74-Hygro-SGFP

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P1093 pCT74-Hygro-SGFP 20μl
P1010 pLenti sgRNA(MS2)-zeo 20μl
P1010 lenti sgRNA(MS2)_zeo 20μl
P0990 pAN8-gpdA 20μl
P0967 p415-PtrpC-Cas9-TtrpC-CYC1t 20μl
P0966 p426-SNR52p-gRNA.csr-1.Y-SUP4t 20μl
P0303 pBARGPE1 20μl "基本信息
启动子: gpdA，TrpC
复制子: pUC ori
终止子: trpC
质粒分类: 广宿主穿梭载体；真菌过表达质粒
质粒大小: 5518bp
原核抗性: 氨苄青霉素Ampicillin
筛选标记: 草胺膦Bar
克隆菌株: DH5α
培养条件: 37℃，有氧，LB
表达宿主: 丝状真菌
诱导方式: 无需诱导
5'测序引物: 根据全序列设计
3'测序引物: 根据全序列设计
备注: 图谱中带下划线的是单一酶切位点
质粒简介
pBARGPE1是一个丝状真菌-大肠杆菌穿梭表达载体，利用gpdA promoter可启动外源目的基因的表达，转化至丝状真菌中可用草胺膦Bar筛选。

质粒图谱

质粒序列
LOCUS       Exported                5518 bp ds-DNA     circular SYN 26-JUL-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pBARGPE1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5518)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported 2015-4-17 from MLCC http://www.miao领bio.com
REFERENCE   2  (bases 1 to 5518)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, July 26, 2016 from SnapGene Viewer 3.1.4
COMMENT     http://www.miao领bio.com/
FEATURES             Location/Qualifiers
     source          1..5518
                     /organism=""synthetic DNA construct""
                     /mol_type=""other DNA""
     misc_feature    complement(49..133)
                     /label=polylinker
                     /note=""polylinker""
     misc_feature    complement(147..831)
                     /label=gpdA Promotor
                     /note=""gpdA Promoter""
                     /note=""gpdA Promotor;promoter region of the A.nidulans gpdA
                     gene""
     misc_feature    complement(1372..1960)
                     /label=pUC origin
                     /note=""pUC origin""
     misc_feature    complement(2131..2991)
                     /label=AmpR
                     /note=""AmpR""
                     /note=""AmpR;beta-lactamase""
     misc_feature    complement(2992..3096)
                     /label=AmpR promoter
                     /note=""AmpR promoter""
     misc_feature    3394..3779
                     /label=TrpC Promotor
                     /note=""TrpC Promotor""
     misc_feature    3780..4331
                     /label=Bar
                     /note=""Bar""
                     /note=""Bar;phosphinothricin acetyltransferase""
     misc_feature    5261..5491
                     /label=trpC terminator
                     /note=""trpC terminator""
                     /note=""trpC terminator;derived from Aspergillus nidulans
                     TrpC""
ORIGIN
        1 ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgatatcga attcctgcag
       61 cccgggggat cctctagagt cgaggtcgac ggtatcgata agcttgatta aaggttcttg
      121 gatgggaaga tgaatatact gaagatggga aaagaaagag aaaagaaaag agcagctggt
      181 ggggagagca ggaaaatatg gcaacaaatg ttggactgac gcaacgacct tgtcaacccc
      241 gccgacacac cgggcggaca gacggggcaa agctgcctac cagggactga gggacctcag
      301 caggtcgagt gcagagcacc ggatgggtcg actgccagct tgtgttcccg gtctgcgccg
      361 ctggccagct cctgagcggc ctttccggtt tcatacaccg ggcaaagcag gagaggcacg
      421 atatttggac gccctacaga tgccggatgg gccaattagg gagcttacgc gccgggtact
      481 cgctctacct acttcggaga aggtactatc tcgtgaatct tttaccagat cggaagcaat
      541 tggacttctg tacctaggtt aatggcatgc tatttcgccg acggctatac acccctggct
      601 tcacattctc cttcgcttac tgccggtgat tcgatgaagc tccatattct ccgatgatgc
      661 aatagattct tggtcaacga ggggcacacc agcctttcca cttcggggcg gaggggcggc
      721 cggtcccgga ttaataatca tccactgcac ctcagagccg ccagagctgt ctggcgcagt
      781 ggcgcttatt actcagccct tctctctgcg tccgtccgtc tctccgcatg ctttgcatac
      841 ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagag
      901 atctctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
      961 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     1021 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
     1081 tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     1141 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc
     1201 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1261 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1321 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1381 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1441 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1501 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1561 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1621 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1681 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1741 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1801 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1861 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1921 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1981 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2041 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2101 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2161 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2221 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     2281 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2341 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2401 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2461 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2521 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2581 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2641 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2701 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     2761 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     2821 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     2881 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     2941 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3001 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3061 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3121 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     3181 gccctttcgt cttcaagaat taattcggtc gaaaaaagaa aaggagaggg ccaagaggga
     3241 gggcattggt gactattgag cacgtgagta tacgtgatta agcacacaaa ggcagcttgg
     3301 agtatgtctg ttattaattt cacaggtagt tctggtccat tggtgaaagt ttgcggcttg
     3361 cagagcacag aggccgcaga atgtgctcta gactcgacag aagatgatat tgaaggagca
     3421 ctttttgggc ttggctggag ctagtggagg tcaacaatga atgcctattt tggtttagtc
     3481 gtccaggcgg tgagcacaaa atttgtgtcg tttgacaaga tggttcattt aggcaactgg
     3541 tcagatcagc cccacttgta gcagtagcgg cggcgctcga agtgtgactc ttattagcag
     3601 acaggaacga ggacattatt atcatctgct gcttggtgca cgataacttg gtgcgtttgt
     3661 caagcaaggt aagtgaacga cccggtcata ccttcttaag ttcgcccttc ctccctttat
     3721 ttcagattca atctgactta cctattctac ccaagcaaag cttcgattag gaagtaacca
     3781 tgagcccaga acgacgcccg gccgacatcc gccgtgccac cgaggcggac atgccggcgg
     3841 tctgcaccat cgtcaaccac tacatcgaga caagcacggt caacttccgt accgagccgc
     3901 aggaaccgca ggagtggacg gacgacctcg tccgtctgcg ggagcgctat ccctggctcg
     3961 tcgccgaggt ggacggcgag gtcgccggca tcgcctacgc gggcccctgg aaggcacgca
     4021 acgcctacga ctggacggcc gagtcgaccg tgtacgtctc cccccgccac cagcggacgg
     4081 gactgggctc cacgctctac acccacctgc tgaagtccct ggaggcacag ggcttcaaga
     4141 gcgtggtcgc tgtcatcggg ctgcccaacg acccgagcgt gcgcatgcac gaggcgctcg
     4201 gatatgcccc ccgcggcatg ctgcgggcgg ccggcttcaa gcacgggaac tggcatgacg
     4261 tgggtttctg gcagctggac ttcagcctgc cggtaccgcc ccgtccggtc ctgcccgtca
     4321 ccgagatctg atccgtcgac ctgcagatcg ttcaaacatt tggcaataaa gtttcttaag
     4381 attgaatcct gttgccggtc ttgcgatgat tatcatataa tttctgttga attacgttaa
     4441 gcatgtaata attaacatgt aatgcatagt accgagaaac tagtggatct aaacataaaa
     4501 tctgtaaaat aacaagatgt aaagataatg ctaaatcatt tggctttttg attgattgta
     4561 caggaaaata tacatcgcag ggggttgact tttaccattt caccgcaatg gaatcaaact
     4621 tgttgaagag aatgttcaca ggcgcatacg ctacaatgac ccgattcttg ctagcctttt
     4681 ctcggtcttg caaacaaccg ccgcggccgg tcgagggacc taataacttc gtatagcata
     4741 cattatacga agttatatta agggttccgg atcgcggccg ctcgacctgc agccaagcta
     4801 gcttggctgg acgtaaactc ctcttcagac ctaataactt cgtatagcat acattatacg
     4861 aagttatatt aagggttatt gaatatgatc ggaattggtc gatcaggtat tgctgttatc
     4921 tgatgagtat acgttgtcct ggccacggca gaagcacgct tatcgctcca atttcccaca
     4981 acattagtca actccgttag gcccttcatt gaaagaaatg aggtcatcaa atgtcttcca
     5041 atgtgagatt ttgggccatt ttttatagca aagattgaat aaggcgcatt tttcttcaaa
     5101 gcttgcaaat taaagccttc gagcgtccca aaaccttctc aagcaaggtt ttcagtataa
     5161 tgttacatgc gtacacgcgc tatattttgt tttctatcgc gtattaaatg tataattgcg
     5221 ggactctaat cataaaaacc catctcataa ataacgtcat gcattgcaga tgagctgtat
     5281 ctggaagagg taaacccgaa acgcgtttta ttcttgttga catggagcta ttaaatcact
     5341 agaaggcact ctttgctgct tggacaaatg aacgtatctt atcgagatcc tgaacaccat
     5401 ttgtctcaac tccggagctg acatcgacac caacgatctt atatccagat tcgtcaagct
     5461 gtttgatgat ttcagtaacg ttaagtggat ctcggtgacg ggcaggaccg gacggggc
//"

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