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
//
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