請在線咨詢或直接聯系17720555249/18986294316

必騰預構載體定制最高100元/個

基本信息

質粒圖譜和多克隆位點信息

注意：

客戶在我公司選擇構建載體服務項目時，請自行提供母源載體。

如要使用本公司保存的母源載體時，本公司不能保證所有母源載體完全符合客戶預期，也不承擔由此產生的任何責任。

pEGFP-C1 encodes a red-shifted variant of wild-type GFP which has been optimized for brighter fluorescence and higher expression in mammalian cells. (Excitation maximum = 488 nm;emission maximum = 507 nm.) pEGFP-C1 encodes the GFPmut1 variant which contains the double-amino-acid substitution of Phe-64 to Leu and Ser-65 to Thr. The coding sequence of the EGFP gene contains more than 190 silent base changes which correspond to human codon-usage preferences. Sequences flanking EGFP have been converted to a Kozak consensus translation initiation site to further increase the translation efficiency in eukaryotic cells. The MCS in pEGFPC1is between the EGFP coding sequences and the SV40 poly A. Genes cloned into the MCS will be expressed as fusions to the C-terminus of EGFP if they are in the same reading frame as EGFP and there are no intervening stop codons. SV40 polyadenylation signals downstream of the EGFP gene direct proper processing of the 3' end of the EGFP mRNA. The vector backbone also contains an SV40 origin for replication in mammalian cells expressing the SV40 T-antigen. A neomycin resistance cassette (Neor), consisting of the SV40 early promoter, the neomycin/kanamycin resistance gene of Tn5, and polyadenylation signals from the Herpes simplex virus thymidine kinase(HSV TK) gene, allows stably transfected eukaryotic cells to be selected using G418. A bacterial promoter upstream of this cassette expresses kanamycin resistance in E.coli. The pEGFP-C1backbone also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single stranded DNA production.         

載體應用

Fusions to the C terminus of EGFP retain the fluorescent properties of the native protein allowing the localization of the fusion protein in vivo. The target gene should be cloned into pEGFP-C1 so that it is in frame with the EGFP coding sequences, with no intervening in-frame stop codons. The recombinant EGFP vector can be transfected into mammalian cells using any standard transfection method. If required, stable transformants can be selected usingG418. pEGFP-C1 can also be used simply to express EGFP in a cell line of interest (e.g., as a transfection marker).

載體序列

LOCUS       Exported                4731 bp ds-DNA     circular SYN 20-MAR-2019

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    .

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 4731)

  AUTHORS   BT-Lab

  TITLE     Direct Submission

  JOURNAL   Exported Wednesday, Mar 20, 2019 from SnapGene Viewer 4.3.4

FEATURES             Location/Qualifiers

     source          1..4731

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     enhancer        61..364

                     /label=CMV enhancer

                     /note="human cytomegalovirus immediate early enhancer"

     promoter        365..568

                     /label=CMV promoter

                     /note="human cytomegalovirus (CMV) immediate early

                     promoter"

     regulatory      607..616

                     /regulatory_class="other"

                     /note="vertebrate consensus sequence for strong initiation

                     of translation (Kozak, 1987)"

     CDS             613..1329

                     /codon_start=1

                     /product="enhanced GFP"

                     /label=EGFP

                     /note="mammalian codon-optimized"

                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL

                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD

                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK

                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL

                     EFVTAAGITLGMDELYK"

     misc_feature    1330..1395

                     /label=MCS

                     /note="multiple cloning site"

     polyA_signal    1519..1640

                     /label=SV40 poly(A) signal

                     /note="SV40 polyadenylation signal"

     rep_origin      complement(1647..2102)

                     /direction=LEFT

                     /label=f1 ori

                     /note="f1 bacteriophage origin of replication; arrow

                     indicates direction of (+) strand synthesis"

     promoter        2129..2233

                     /gene="bla"

                     /label=AmpR promoter

     promoter        2235..2592

                     /label=SV40 promoter

                     /note="SV40 enhancer and early promoter"

     rep_origin      2443..2578

                     /label=SV40 ori

                     /note="SV40 origin of replication"

     CDS             2627..3421

                     /codon_start=1

                     /gene="aph(3')-II (or nptII)"

                     /product="aminoglycoside phosphotransferase from Tn5"

                     /label=NeoR/KanR

                     /note="confers resistance to neomycin, kanamycin, and G418

                     (Geneticin(R))"

                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP

                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS

                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ

                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA

                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"

     polyA_signal    3653..3700

                     /label=HSV TK poly(A) signal

                     /note="herpes simplex virus thymidine kinase

                     polyadenylation signal (Cole and Stacy, 1985)"

     rep_origin      4029..4617

                     /direction=RIGHT

                     /label=ori

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of

                     replication"

ORIGIN

        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg

       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt

      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca

      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc

      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta

      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac

      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg

      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg

      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt

      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta

      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg

      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc

      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg

      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc

      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag

      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag

      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac

     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac

     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc

     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg

     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc

     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag

     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc

     1381 gcgggcccgg gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta

     1441 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg

     1501 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat

     1561 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc

     1621 aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc

     1681 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc

     1741 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag

     1801 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga

     1861 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc

     1921 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa

     1981 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt

     2041 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc

     2101 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat

     2161 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg

     2221 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt

     2281 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca

     2341 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat

     2401 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg

     2461 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc

     2521 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta

     2581 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga

     2641 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc

     2701 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc

     2761 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc

     2821 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac

     2881 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc

     2941 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac

     3001 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg

     3061 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct

     3121 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt

     3181 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg

     3241 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac

     3301 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg

     3361 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg

     3421 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat

     3481 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc

     3541 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg

     3601 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata

     3661 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc

     3721 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt

     3781 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac

     3841 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta

     3901 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc

     3961 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa

     4021 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca

     4081 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta

     4141 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc

     4201 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca

     4261 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta

     4321 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag

     4381 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt

     4441 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc

     4501 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac

     4561 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac

     4621 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc

     4681 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca t

//