P0382 pBARGPE1-mCherry

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P0382 pBARGPE1-mCherry 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: 6229bp
质粒标签: 单体红色荧光蛋白mCherry
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
质粒简介
pBARGPE1-mCherry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                6229 bp ds-DNA     circular SYN 30-AUG-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pBARGPE1-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6229)
  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..6229
                     /organism=""synthetic DNA construct""
                     /mol_type=""other DNA""
     misc_feature    28..258
                     /note=""trpC terminator""
     protein_bind    complement(652..685)
                     /bound_moiety=""Cre recombinase""
                     /note=""loxP""
                     /note=""Cre-mediated recombination occurs in the 8-bp core
                     sequence (GCATACAT).""
     protein_bind    complement(763..796)
                     /bound_moiety=""Cre recombinase""
                     /note=""loxP""
                     /note=""Cre-mediated recombination occurs in the 8-bp core
                     sequence (GCATACAT).""
     CDS             complement(1188..1739)
                     /codon_start=1
                     /gene=""Streptomyces hygroscopicus bar""
                     /product=""phosphinothricin acetyltransferase""
                     /note=""BlpR""
                     /note=""confers resistance to bialophos or phosphinothricin""
                     /translation=""MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI""
     misc_feature    complement(1740..2125)
                     /note=""TrpC Promotor""
     promoter        complement(2216..2317)
                     /gene=""S. cerevisiae TRP1""
                     /note=""TRP1 promoter""
     promoter        2423..2527
                     /gene=""bla""
                     /note=""AmpR promoter""
     CDS             2528..3388
                     /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""
     rep_origin      3559..4147
                     /direction=RIGHT
                     /note=""ori""
                     /note=""high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication""
     misc_feature    4333..4473
                     /note=""bom""
                     /note=""basis of mobility region from pBR322""
     misc_feature    4688..5372
                     /note=""gpdA Promoter""
     primer_bind     complement(5422..5438)
                     /note=""KS primer""
                     /note=""common sequencing primer, one of multiple similar
                     variants""
     CDS             5474..6184
                     /codon_start=1
                     /product=""monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)""
                     /note=""mCherry""
                     /note=""mammalian codon-optimized""
                     /translation=""MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK""
     primer_bind     complement(6196..6212)
                     /note=""KS primer""
                     /note=""common sequencing primer, one of multiple similar
                     variants""
ORIGIN
        1 gccccgtccg gtcctgcccg tcaccgagat ccacttaacg ttactgaaat catcaaacag
       61 cttgacgaat ctggatataa gatcgttggt gtcgatgtca gctccggagt tgagacaaat
      121 ggtgttcagg atctcgataa gatacgttca tttgtccaag cagcaaagag tgccttctag
      181 tgatttaata gctccatgtc aacaagaata aaacgcgttt cgggtttacc tcttccagat
      241 acagctcatc tgcaatgcat gacgttattt atgagatggg tttttatgat tagagtcccg
      301 caattataca tttaatacgc gatagaaaac aaaatatagc gcgtgtacgc atgtaacatt
      361 atactgaaaa ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgcaagctt
      421 tgaagaaaaa tgcgccttat tcaatctttg ctataaaaaa tggcccaaaa tctcacattg
      481 gaagacattt gatgacctca tttctttcaa tgaagggcct aacggagttg actaatgttg
      541 tgggaaattg gagcgataag cgtgcttctg ccgtggccag gacaacgtat actcatcaga
      601 taacagcaat acctgatcga ccaattccga tcatattcaa taacccttaa tataacttcg
      661 tataatgtat gctatacgaa gttattaggt ctgaagagga gtttacgtcc agccaagcta
      721 gcttggctgc aggtcgagcg gccgcgatcc ggaaccctta atataacttc gtataatgta
      781 tgctatacga agttattagg tccctcgacc ggccgcggcg gttgtttgca agaccgagaa
      841 aaggctagca agaatcgggt cattgtagcg tatgcgcctg tgaacattct cttcaacaag
      901 tttgattcca ttgcggtgaa atggtaaaag tcaaccccct gcgatgtata ttttcctgta
      961 caatcaatca aaaagccaaa tgatttagca ttatctttac atcttgttat tttacagatt
     1021 ttatgtttag atccactagt ttctcggtac tatgcattac atgttaatta ttacatgctt
     1081 aacgtaattc aacagaaatt atatgataat catcgcaaga ccggcaacag gattcaatct
     1141 taagaaactt tattgccaaa tgtttgaacg atctgcaggt cgacggatca gatctcggtg
     1201 acgggcagga ccggacgggg cggtaccggc aggctgaagt ccagctgcca gaaacccacg
     1261 tcatgccagt tcccgtgctt gaagccggcc gcccgcagca tgccgcgggg ggcatatccg
     1321 agcgcctcgt gcatgcgcac gctcgggtcg ttgggcagcc cgatgacagc gaccacgctc
     1381 ttgaagccct gtgcctccag ggacttcagc aggtgggtgt agagcgtgga gcccagtccc
     1441 gtccgctggt ggcgggggga gacgtacacg gtcgactcgg ccgtccagtc gtaggcgttg
     1501 cgtgccttcc aggggcccgc gtaggcgatg ccggcgacct cgccgtccac ctcggcgacg
     1561 agccagggat agcgctcccg cagacggacg aggtcgtccg tccactcctg cggttcctgc
     1621 ggctcggtac ggaagttgac cgtgcttgtc tcgatgtagt ggttgacgat ggtgcagacc
     1681 gccggcatgt ccgcctcggt ggcacggcgg atgtcggccg ggcgtcgttc tgggctcatg
     1741 gttacttcct aatcgaagct ttgcttgggt agaataggta agtcagattg aatctgaaat
     1801 aaagggagga agggcgaact taagaaggta tgaccgggtc gttcacttac cttgcttgac
     1861 aaacgcacca agttatcgtg caccaagcag cagatgataa taatgtcctc gttcctgtct
     1921 gctaataaga gtcacacttc gagcgccgcc gctactgcta caagtggggc tgatctgacc
     1981 agttgcctaa atgaaccatc ttgtcaaacg acacaaattt tgtgctcacc gcctggacga
     2041 ctaaaccaaa ataggcattc attgttgacc tccactagct ccagccaagc ccaaaaagtg
     2101 ctccttcaat atcatcttct gtcgagtcta gagcacattc tgcggcctct gtgctctgca
     2161 agccgcaaac tttcaccaat ggaccagaac tacctgtgaa attaataaca gacatactcc
     2221 aagctgcctt tgtgtgctta atcacgtata ctcacgtgct caatagtcac caatgccctc
     2281 cctcttggcc ctctcctttt cttttttcga ccgaattaat tcttgaagac gaaagggcct
     2341 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     2401 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     2461 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     2521 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     2581 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     2641 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     2701 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     2761 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     2821 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     2881 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     2941 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     3001 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     3061 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     3121 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     3181 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     3241 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     3301 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     3361 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     3421 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     3481 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3541 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3601 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3661 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3721 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3781 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3841 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3901 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3961 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac