• 规格：10μl质粒
• 启动子：T7 promoter
• 复制子：ColE1 ori
• 终止子：T7 terminator
• 质粒大小：4640bp
• 质粒标签：N-T7
• 原核抗性：氨苄青霉素Amp
• 克隆菌株：大肠杆菌DH5α
• 表达宿主：大肠杆菌BL21(DE3)
• 诱导方式：无需诱导
• 5测序引物：T7:TAATACGACTCACTATAGGG
• 3测序引物：T7-ter:TGCTAGTTATTGCTCAGCGG
• 备注：组成型表达
• 质粒简介：         pET-3a载体是一个大肠杆菌表达载体，T7启动子驱动T7标签和目的基因融合表达。该载体是组成型表达，并且不携带6×HIS标签。The pET-3a-d vectors carry an N-terminal T7•Tag sequence and BamH I cloning site. These vectors are the precursors to many pET family vectors; the pET-23a-d(+) series corresponds to pET-3a-d but incorporates several additional features. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circular map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below.
• 质粒图谱1： png/1-1G02G016255Z.png

质粒序列： 点击查看序列

LOCUS       Exported                4640 bp ds-DNA     circular SYN 28-7-2015
DEFINITION  synthetic circular DNA
KEYWORDS    Untitled
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4640)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported 2015-7-28 from SnapGene Viewer 2.8.0
            http://www.fenghbio.cn
FEATURES             Location/Qualifiers
     source          1..4640
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          559..581
                     /organism="Enterobacteria phage T7"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:10760"
     promoter        10..38
                     /note="tet promoter"
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     terminator      404..451
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     CDS             complement(519..551)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /note="T7 tag (gene 10 leader)"
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     RBS             559..581
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     promoter        complement(613..631)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2192..2383
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    2485..2627
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2813..3401)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(3572..4432)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4433..4537)
                     /gene="bla"
                     /note="AmpR promoter"
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata
      241 tgcgttgatg caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg
      301 ccgcccagtc ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac
      361 cacacccgtc ctgtggatat ccggatatag ttcctccttt cagcaaaaaa cccctcaaga
      421 cccgtttaga ggccccaagg ggttatgcta gttattgctc agcggtggca gcagccaact
      481 cagcttcctt tcgggctttg ttagcagccg gatccgcgac ccatttgctg tccaccagtc
      541 atgctagcca tatgtatatc tccttcttaa agttaaacaa aattatttct agagggaaac
      601 cgttgtggtc tccctatagt gagtcgtatt aatttcgcgg gatcgagatc tcgatcctct
      661 acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata
      721 tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg agcgcttgtt
      781 tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgcc atctccttgc
      841 atgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg ggctgcttcc
      901 taatgcagga gtcgcataag ggagagcgtc gaccgatgcc cttgagagcc ttcaacccag
      961 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     1021 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc
     1081 gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg
     1141 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca
     1201 ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag
     1261 gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt
     1321 tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt caaggatcgc
     1381 tcgcggctct taccagccta acttcgatca ctggaccgct gatcgtcacg gcgatttatg
     1441 ccgcctcggc gagcacatgg aacgggttgg catggattgt aggcgccgcc ctataccttg
     1501 tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc tgaatggaag
     1561 ccggcggcac ctcgctaacg gattcaccac tccaagaatt ggagccaatc aattcttgcg
     1621 gagaactgtg aatgcgcaaa ccaacccttg gcagaacata tccatcgcgt ccgccatctc
     1681 cagcagccgc acgcggcgca tctcgggcag cgttgggtcc tggccacggg tgcgcatgat
     1741 cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa
     1801 tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg
     1861 agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc
     1921 agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg
     1981 aacacctaca tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc
     2041 ccgccgcatc cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc
     2101 atcatcagta acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat
     2161 gaacagaaat cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa
     2221 catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga
     2281 cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg
     2341 cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     2401 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     2461 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     2521 gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatatgc
     2581 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     2641 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2701 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2761 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2821 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2881 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2941 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3001 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3061 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3121 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3181 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3241 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3301 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3361 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3421 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3481 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3541 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3601 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3661 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3721 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3781 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3841 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg
     3901 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3961 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4021 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4081 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4141 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata
     4201 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4261 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4321 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4381 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4441 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4501 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4561 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     4621 ggccctttcg tcttcaagaa
//