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产品名称：pTracer-CMV2
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简单介绍

pTracer-CMV2启动子后有MCS供插入外源基因，hEF1α启动子调控Cycle 3 GFP-Zeocin fusion的表达，可以做到既有荧光蛋白来做转染指示，且荧光基因表达和外源基因表达又不会互相干扰，而且Zeocin也��以用来做稳定性细胞筛选。pTracer-CMV2另外，由于EM-7启动子的存在，Cycle 3 GFP-Zeocin fusion在E.coli中也可以表达，可以用来做原核荧光定位。

产品描述

pTracer-CMV2

产品信息
基本信息
质粒简介
质粒图谱
质粒序列

产品信息

产品货号	产品名称	产品规格	优惠价
P0169	pTracer-CMV2	5μg	¥请询价

使用说明

信裕质粒平台的各批次质粒菌株发货前均经过严格的多重验证，如存在质量问题，请在收到产品的三个月内通知我司。收到质粒后请短暂离心，加入20μl ddH2O溶解质粒，取2μl转化至对应感受态中，挑取单克隆重新提取质粒后使用。如需获取其他详细信息请登录我司官网查询。

基本信息

启动子:	CMV promoter
复制子:	pUC ori
终止子:	bGH poly(A) signal
质粒分类:	哺乳系列质粒；哺乳表达质粒；pCMV系列质粒
质粒大小:	6210bp
原核抗性:	氨苄青霉素Amp（100μg/ml）、博来霉素Zeocin（50μg/ml）
筛选标记:	博来霉素Zeocin
克隆菌株:	DH5α等大肠杆菌
培养条件:	37℃，有氧，LB
表达宿主:	293T等哺乳细胞
培养条件:	37℃，5%CO2
诱导方式:	无需诱导
5'测序引物:	CMV-F（CGCAAATGGGCGGTAGGCGTG）
3'测序引物:	BGH（TAGAAGGCACAGTCGAGG）

质粒简介

pTracer-CMV2的CMV启动子后有MCS供插入外源基因，hEF1α启动子调控Cycle 3 GFP-Zeocin fusion的表达，可以做到既有荧光蛋白来做转染指示，且荧光基因表达和外源基因表达又不会互相干扰，而且Zeocin也可以用来做稳定性细胞筛选。另外，由于EM-7启动子的存在，Cycle 3 GFP-Zeocin fusion在E.coli中也可以表达，可以用来做原核荧光定位。
  pTracer-CMV2 is a 6.2 kb mammalian expression vector derived from pcDNA3.1 and designed for visual detection of transformed E. coli cells and transfected mammalian cells. The vector contains the following elements:
  Cycle 3-GFP, an improved GFP (Green Fluorescent Protein) gene (Crameri et al.,1996), fused to the Zeocin™ resistance gene (see below) for convenient,noninvasive detection of transformed or transfected cells
  • Human elongation factor 1-subunit promoter (hEF-1) for mammalian expression of the Cycle 3-GFP-Zeocin fusion (Goldman et al., 1996; Mizushima and Nagata, 1990)
  • A synthetic bacterial promoter, EM-7, for expression of Cycle 3-GFP in E. coli
  • Human cytomegalovirus immediate-early (CMV) promoter for high-level expression of your gene in a wide range of mammalian cells
  • Zeocin™ resistance for stable selection in mammalian cell lines
  • Ampicillin and Zeocin resistance genes for selection of transformants in E. coli
  The Cycle 3-GFP gene used in this vector is described in Crameri et al., 1996. In this paper, the codon usage was optimized for expression in mammalian cells and three cycles of DNA shuffling were used to generate a mutant form of GFP that has the following characteristics:
  • Excitation and emission maxima that are the same as wild-type GFP (395 nm and 478 nm for primary and secondary excitation, respectively, and 507 nm for emission)
  • High solubility in E. coli for visual detection of transformed cells
  • >40-fold increase in fluorescent yield over wild-type GFP
  • The Cycle 3-GFP gene is fused to the Zeocin™ resistance marker to correlate GFP fluorescence with resistance to the antibiotic Zeocin.
  Zeocin is a member of the bleomycin/phleomycin family of antibiotics isolated from Streptomyces. It shows strong toxicity against bacteria, fungi (including yeast), plants, and mammalian cell lines. It is particularly well-suited for selection of mammalian stable cell lines. Ordering information is provided below.

质粒图谱

质粒序列

LOCUS       Exported                6210 bp ds-DNA    circular SYN 04-11-2015
KEYWORDS    Untitled 4
SOURCE      synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE   1 (bases 1 to 6210)
AUTHORS   admin
TITLE     Direct Submission
JOURNAL   Exported 2015-11-4   from MiaoLingPlasmid
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..6210
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        235..614
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1028..1139
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     promoter        1309..2487
                     /note="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     intron          1540..2478
                     /note="EF-1-alpha intron A"
                     /note="intron upstream of the start codon of human
                     EF-1-alpha"
     promoter        2504..2522
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2611..2658
                     /note="EM7 promoter"
                     /note="synthetic bacterial promoter "
     CDS             3376..3753
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /note="BleoR"
                     /note="confers resistance to bleomycin, phleomycin, and
                     Zeocin(TM)"
                     /translation="MDAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRD
                     DVTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWG
                     REFALRDPAGNCVHFVAEEQD"
     polyA_signal    3883..4004
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4053..4069)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    4077..4093
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4101..4131)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    4146..4167
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4455..5043)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5214..6074)
                     /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(6075..6179)
                     /gene="bla"
                     /note="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc
      961 agatatccag cacagtggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca
     1021 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     1081 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     1141 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     1201 gaggattggg aagacaatag cagctttgca aagatggata aagttttaaa cagagaggaa
     1261 tctttgcagc taatggacct tctaggtctt gaaaggagtg cctcgtgagg ctccggtgcc
     1321 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
     1381 aattgaaccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
     1441 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     1501 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc
     1561 cgcgggcctg gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccacctgg
     1621 ctgcagtacg tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg
     1681 ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg
     1741 ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct
     1801 ctagccattt aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt
     1861 gtaaatgcgg gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac
     1921 ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg
     1981 agaatcggac gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg
     2041 ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg
     2101 gaaagatggc cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg
     2161 ggagagcggg cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc
     2221 gcttcatgtg actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc
     2281 ttttggagta cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac
     2341 actgagtggg tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa
     2401 tttgcccttt ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt
     2461 ttttttcttc catttcaggt gtcgtgagga attagcttgg tactaatacg actcactata
     2521 gggagaccca agctggctag gtaagctcct aggcttttgc aaaaagctcc cgggagcttg
     2581 tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc atagtatatc
     2641 ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ctagcaaagg agaagaactt
     2701 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
     2761 tctgtcagtg gagagggtga aggtgatgct acatacggaa agcttaccct taaatttatt
     2821 tgcactactg gaaaactacc tgttccatgg ccaacacttg tcactacttt ctcttatggt
     2881 gttcaatgct tttcccgtta tccggatcat atgaaacggc atgacttttt caagagtgcc
     2941 atgcccgaag gttatgtaca ggaacgcact atatctttca aagatgacgg gaactacaag
     3001 acgcgtgctg aagtcaagtt tgaaggtgat acccttgtta atcgtatcga gttaaaaggt
     3061 attgatttta aagaagatgg aaacattctc ggacacaaac tcgagtacaa ctataactca
     3121 cacaatgtat acatcacggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt
     3181 cgccacaaca ttgaagatgg atccgttcaa ctagcagacc attatcaaca aaatactcca
     3241 attggcgatg gccctgtcct tttaccagac aaccattacc tgtcgacaca atctgccctt
     3301 tcgaaagatc ccaacgaaaa gcgtgaccac atggtccttc ttgagtttgt aactgctgct
     3361 gggattacac atggcatgga tgccaagttg accagtgccg ttccggtgct caccgcgcgc
     3421 gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg
     3481 gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag
     3541 gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg
     3601 tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg
     3661 accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac
     3721 tgcgtgcact tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc
     3781 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     3841 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     3901 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     3961 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     4021 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     4081 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     4141 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     4201 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4261 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4321 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4381 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4441 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4501 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4561 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4621 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     4681 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4741 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4801 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4861 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     4921 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4981 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5041 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5101 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5161 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5221 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5281 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5341 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5401 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5461 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5521 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5581 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5641 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5701 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5761 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5821 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5881 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5941 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6001 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6061 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6121 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6181 acatttcccc gaaaagtgcc acctgacgtc
//

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产品目录

产品资料

pTracer-CMV2

pTracer-CMV2

产品信息

使用说明

基本信息

质粒简介

质粒图谱

质粒序列