P0407 pBARGPE1-Hygro-EGFP

P0407 pBARGPE1-Hygro-EGFP 20μl "启动子: gpdA，TrpC
复制子: pUC ori
终止子: trpC
质粒分类: 其他宿主穿梭载体；丝状真菌表达质粒
质粒大小: 6700bp
质粒标签: 增强型绿色荧光蛋白EGFP
原核抗性: 氨苄青霉素Ampicillin
筛选标记: 潮霉素Hygromycin
克隆菌株: DH5α
培养条件: 37℃，有氧，LB
表达宿主: 丝状真菌
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: 可表达EGFP融合蛋白
质粒简介
pBARGPE1-Hygro-EGFP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潮霉素Hygromycin筛选。

质粒图谱

质粒序列
LOCUS       Exported                6700 bp ds-DNA     circular SYN 30-AUG-2016
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pBARGPE1-Hygro-EGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6700)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, August 30, 2016 from SnapGene Viewer 3.1.4
            http://www.miao领bio.com
FEATURES             Location/Qualifiers
     source          1..6700
                     /organism=""synthetic DNA construct""
                     /mol_type=""other DNA""
     primer_bind     18..34
                     /note=""KS primer""
                     /note=""common sequencing primer, one of multiple similar
                     variants""
     CDS             complement(55..774)
                     /codon_start=1
                     /product=""enhanced GFP""
                     /note=""EGFP""
                     /note=""mammalian codon-optimized""
                     /translation=""MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK""
     primer_bind     789..805
                     /note=""KS primer""
                     /note=""common sequencing primer, one of multiple similar
                     variants""
     misc_feature    complement(855..1539)
                     /note=""gpdA Promotor""
     misc_feature    1754..1894
                     /note=""bom""
                     /note=""basis of mobility region from pBR322""
     rep_origin      complement(2080..2668)
                     /direction=LEFT
                     /note=""ori""
                     /note=""high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication""
     CDS             complement(2839..3699)
                     /codon_start=1
                     /gene=""bla""
                     /product=""beta-lactamase""
                     /note=""AmpR""
                     /note=""confers resistance to ampicillin, carbenicillin, and
                     related antibiotics""
                     /translation=""MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW""
     promoter        complement(3700..3804)
                     /gene=""bla""
                     /note=""AmpR promoter""
     promoter        3910..4011
                     /gene=""S. cerevisiae TRP1""
                     /note=""TRP1 promoter""
     misc_feature    4102..4487
                     /note=""TrpC Promotor""
     CDS             4488..5513
                     /codon_start=1
                     /gene=""aph(4)-Ia""
                     /product=""aminoglycoside phosphotransferase from E. coli""
                     /note=""HygR""
                     /note=""confers resistance to hygromycin""
                     /translation=""MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K""
     protein_bind    5905..5938
                     /bound_moiety=""Cre recombinase""
                     /note=""loxP""
                     /note=""Cre-mediated recombination occurs in the 8-bp core
                     sequence (GCATACAT).""
     protein_bind    6016..6049
                     /bound_moiety=""Cre recombinase""
                     /note=""loxP""
                     /note=""Cre-mediated recombination occurs in the 8-bp core
                     sequence (GCATACAT).""
     misc_feature    6443..6673
                     /note=""trpC terminator""
ORIGIN
        1 ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgatatcga attcttactt
       61 gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat
      121 gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac tgggtgctca ggtagtggtt
      181 gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag
      241 ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga tgccgttctt
      301 ctgcttgtcg gccatgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc
      361 caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt
      421 gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat
      481 ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat
      541 gtggtcgggg tagcggctga agcactgcac gccgtaggtc agggtggtca cgagggtggg
      601 ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccgtaggt
      661 ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag
      721 ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatggatcc
      781 tctagagtcg aggtcgacgg tatcgataag cttgattaaa ggttcttgga tgggaagatg
      841 aatatactga agatgggaaa agaaagagaa aagaaaagag cagctggtgg ggagagcagg
      901 aaaatatggc aacaaatgtt ggactgacgc aacgaccttg tcaaccccgc cgacacaccg
      961 ggcggacaga cggggcaaag ctgcctacca gggactgagg gacctcagca ggtcgagtgc
     1021 agagcaccgg atgggtcgac tgccagcttg tgttcccggt ctgcgccgct ggccagctcc
     1081 tgagcggcct ttccggtttc atacaccggg caaagcagga gaggcacgat atttggacgc
     1141 cctacagatg ccggatgggc caattaggga gcttacgcgc cgggtactcg ctctacctac
     1201 ttcggagaag gtactatctc gtgaatcttt taccagatcg gaagcaattg gacttctgta
     1261 cctaggttaa tggcatgcta tttcgccgac ggctatacac ccctggcttc acattctcct
     1321 tcgcttactg ccggtgattc gatgaagctc catattctcc gatgatgcaa tagattcttg
     1381 gtcaacgagg ggcacaccag cctttccact tcggggcgga ggggcggccg gtcccggatt
     1441 aataatcatc cactgcacct cagagccgcc agagctgtct ggcgcagtgg cgcttattac
     1501 tcagcccttc tctctgcgtc cgtccgtctc tccgcatgct ttgcatactt ctgcctgctg
     1561 gggagcctgg ggactttcca caccctaact gacacacatt ccacagagat ctctgcctcg
     1621 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     1681 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     1741 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct
     1801 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     1861 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga
     1921 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1981 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     2041 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     2101 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     2161 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     2221 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     2281 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     2341 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     2401 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     2461 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     2521 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2581 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2641 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2701 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2761 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2821 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2881 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     2941 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3001 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3061 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3121 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3181 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3241 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3301 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     3361 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     3421 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     3481 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3541 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3601 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3661 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3721 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3781 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     3841 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     3901 tcaagaatta attcggtcga aaaaagaaaa ggagagggcc aagagggagg gcattggtga
     3961 ctattgagca cgtgagtata cgtgattaag cacacaaagg cagcttggag tatgtctgtt
     4021 attaatttca caggtagttc tggtccattg gtgaaagttt gcggcttgca gagcacagag
     4081 gccgcagaat gtgctctaga ctcgacagaa gatgatattg aaggagcact ttttgggctt​