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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。

載體描述

pGenesil-1質粒是以pEGFP-C1質粒為骨架改造獲得的。pEGFP-C1質粒廣泛用于哺乳動物細胞已經多年，它具有熒光蛋白表達，非常方便導入效果的檢測。

（1） 調控shRNA的啟動子為hU6啟動子。shRNA插入位點：BamHI、HindIII; 

（2） 此載體帶有EGFP熒光基因，可作為轉染指示; 

（3） 此載體帶有Neo抗性基因，可在真核細胞中用G418進行篩選。

 載體序列

LOCUS       Exported                4857 bp ds-DNA     circular SYN 04-MAR-2019

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    .

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 4857)

  AUTHORS   .

  TITLE     Direct Submission

  JOURNAL   Exported Monday, Mar 4, 2019 from SnapGene Viewer 4.3.4

 FEATURES             Location/Qualifiers

     source          1..4857

                     /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"

     primer_bind     1266..1287

                     /label=pEGFP-C-5'

     misc_feature    1476..1597

                     /label=SV40 polyA

                     /note="PolyA"

     misc_feature    1615..1639

                     /label=shRNA

     promoter        complement(1649..1889)

                     /label=U6 promoter

                     /note="RNA polymerase III promoter for human U6 snRNA"

     primer_bind     complement(1699..1721)

                     /label=U6-5'

     promoter        2255..2359

                     /gene="bla"

                     /label=AmpR promoter

     promoter        2361..2718

                     /label=SV40 promoter

                     /note="SV40 enhancer and early promoter"

     rep_origin      2569..2704

                     /label=SV40 ori

                     /note="SV40 origin of replication"

     CDS             2753..3547

                     /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    3779..3826

                     /label=HSV TK poly(A) signal

                     /note="herpes simplex virus thymidine kinase

                     polyadenylation signal (Cole and Stacy, 1985)"

     rep_origin      4155..4743

                     /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 atctcgagat ccaccggatc tagataactg atcataatca

     1381 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga

     1441 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg

     1501 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt

     1561 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtcccggga agcttgcatg

     1621 cctgcaggtc gactctagag gatcccgcgt cctttccaca agatatataa acccaagaaa

     1681 tcgaaatact ttcaagttac ggtaagcata tgatagtcca ttttaaaaca taattttaaa

     1741 actgcaaact acccaagaaa ttattacttt ctacgtcacg tattttgtac taatatcttt

     1801 gtgtttacag tcaaattaat tctaattatc tctctaacag ccttgtatcg tatatgcaaa

     1861 tatgaaggaa tcatgggaaa taggccctct tcctgcccga ccttggcgcg cgctcggcgc

     1921 gcggtcacgc tccgtcacgt ggtgcgtttt gcctgcgcgt ctttccactg ggggaattcg

     1981 tcgactctag acactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg

     2041 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc

     2101 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc

     2161 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca

     2221 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt

     2281 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata

     2341 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta

     2401 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat

     2461 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc

     2521 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta

     2581 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca

     2641 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga

     2701 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga

     2761 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga

     2821 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg

     2881 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga

     2941 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt

     3001 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct

     3061 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct

     3121 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg

     3181 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca

     3241 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga

     3301 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt

     3361 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt

     3421 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct

     3481 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt

     3541 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca

     3601 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg

     3661 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct

     3721 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg

     3781 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt

     3841 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc

     3901 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca

     3961 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt

     4021 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc

     4081 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag

     4141 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa

     4201 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg

     4261 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag

     4321 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg

     4381 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga

     4441 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc

     4501 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc

     4561 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga

     4621 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt

     4681 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg

     4741 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac

     4801 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat

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