pBARGPE1-Hygro-EGFP真菌表达质粒

基本信息

启动子:	gpdA promoter
复制子:	pUC ori
终止子:	trpC terminator
质粒分类:	丝状真菌绿色荧光表达质粒
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:	大肠杆菌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

质粒简介
       pBARGPE1-Hygro-EGFP是一个大肠杆菌-丝状真菌穿梭绿色荧光表达质粒，转化至大肠杆菌可用氨苄抗生素（终浓度100ug/ml）筛选，转化至丝状真菌中可用潮霉素Hygromycin（具体浓度请先做预实验测试或者参考相关文献）筛选，在真菌中可利用gpdA promoter启动绿色荧光蛋白EGFP的表达。因为本质粒的EGFP携带有终止密码子，所以不方便再克隆外源目的基因进入，如需要克隆外源基因请联系本平公司制适合您实验的载体。质粒图谱
质粒序列
LOCUS       Exported                6700 bp ds-DNA     circular SYN 15-AUG-2018
DEFINITION  synthetic circular DNA
KEYWORDS    pBARGPE1-Hygro-EGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6700)
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6700
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(55..774)
                     /codon_start=1
                     /product="enhanced GFP"
                     /note="EGFP"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    complement(854..1539)
                     /note="gpdA Promoter"
     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"
     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    6442..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
     4141 ggctggagct agtggaggtc aacaatgaat gcctattttg gtttagtcgt ccaggcggtg
     4201 agcacaaaat ttgtgtcgtt tgacaagatg gttcatttag gcaactggtc agatcagccc
     4261 cacttgtagc agtagcggcg gcgctcgaag tgtgactctt attagcagac aggaacgagg
     4321 acattattat catctgctgc ttggtgcacg ataacttggt gcgtttgtca agcaaggtaa
     4381 gtgaacgacc cggtcatacc ttcttaagtt cgcccttcct ccctttattt cagattcaat
     4441 ctgacttacc tattctaccc aagcaaagct tcgattagga agtaaccatg aaaaagcctg
     4501 aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc
     4561 tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg
     4621 gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc
     4681 ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg gagtttagcg
     4741 agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg
     4801 aaaccgaact gcccgctgtt ctacaaccgg tcgcggaggc tatggatgcg atcgctgcgg
     4861 ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca
     4921 ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg
     4981 tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg
     5041 ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc
     5101 tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt
     5161 cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc
     5221 agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccacga ctccgggcgt
     5281 atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg
     5341 atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg
     5401 ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac
     5461 tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaagaaa tagtccgtcg
     5521 acctgcagat cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg
     5581 tcttgcgatg attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat
     5641 gtaatgcata gtaccgagaa actagtggat ctaaacataa aatctgtaaa ataacaagat
     5701 gtaaagataa tgctaaatca tttggctttt tgattgattg tacaggaaaa tatacatcgc
     5761 agggggttga cttttaccat ttcaccgcaa tggaatcaaa cttgttgaag agaatgttca
     5821 caggcgcata cgctacaatg acccgattct tgctagcctt ttctcggtct tgcaaacaac
     5881 cgccgcggcc ggtcgaggga cctaataact tcgtatagca tacattatac gaagttatat
     5941 taagggttcc ggatcgcggc cgctcgacct gcagccaagc tagcttggct ggacgtaaac
     6001 tcctcttcag acctaataac ttcgtatagc atacattata cgaagttata ttaagggtta
     6061 ttgaatatga tcggaattgg tcgatcaggt attgctgtta tctgatgagt atacgttgtc
     6121 ctggccacgg cagaagcacg cttatcgctc caatttccca caacattagt caactccgtt
     6181 aggcccttca ttgaaagaaa tgaggtcatc aaatgtcttc caatgtgaga ttttgggcca
     6241 ttttttatag caaagattga ataaggcgca tttttcttca aagcttgcaa attaaagcct
     6301 tcgagcgtcc caaaaccttc tcaagcaagg ttttcagtat aatgttacat gcgtacacgc
     6361 gctatatttt gttttctatc gcgtattaaa tgtataattg cgggactcta atcataaaaa
     6421 cccatctcat aaataacgtc atgcattgca gatgagctgt atctggaaga ggtaaacccg
     6481 aaacgcgttt tattcttgtt gacatggagc tattaaatca ctagaaggca ctctttgctg
     6541 cttggacaaa tgaacgtatc ttatcgagat cctgaacacc atttgtctca actccggagc
     6601 tgacatcgac accaacgatc ttatatccag attcgtcaag ctgtttgatg atttcagtaa
     6661 cgttaagtgg atctcggtga cgggcaggac cggacggggc
//