基本信息
| 启动子: | CMV promoter |
| 复制子: | pUC ori |
| 终止子: | SV40 poly(A) signal |
| 质粒大小: | 9237bp |
| 原核抗性: | Kan |
| 筛选标记: | Neo |
| 克隆菌株: | Stbl3 |
| 培养条件: | 37℃，有氧，LB |
| 诱导方式: | 无须诱导，瞬时表达 |
质粒简介
PAdTrack-CMV transgenic adenovirus vector to cell expression vector with CMV promoter and EGFP gene expression of green fluorescent protein tracking results, and use common pAdeasy1 vector maximum can be inserted into the 5.9kb, the use of common pAdeasy2 carrier can insert the 8.6kb. The specific use of the method can refer to the pAdeasy carrier system.
Recombinant adenoviruses provide a versatile system for gene expression studies and therapeutic applications. We here report a strategy which simplifies the generation and production of such viruses. A recombinant adenoviral plasmid is generated with a minimum of enzymatic manipulations, employing homologous recombination in bacteria rather than in eucaryotic cells. Following transfections of such plasmids into a mammalian packaging cell line, viral production is conveniently followed with the aid of green fluorescent protein, encoded by a gene incorporated into the viral backbone. Homogeneous viruses can be obtained from this procedure without plaque purification.This system should expedite the process of generating and testing recombinant adenoviruses for a variety of purposes.质粒图谱
质粒序列
LOCUS Exported 9237 bp ds-DNA circular SYN 27-AUG-2016
DEFINITION synthetic circular DNA
KEYWORDS Untitled 3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9237)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..9237
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 79..181
/note="ITR"
/note="inverted terminal repeat of human adenovirus
serotype 5"
enhancer 3440..3743
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 3744..3947
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
polyA_signal 4158..4279
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
enhancer 4365..4668
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 4669..4872
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 4917..5633
/codon_start=1
/product="enhanced GFP"
/note="EGFP"
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
polyA_signal 5772..5893
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
misc_signal 6036..6186
/note="Ad5 Psi"
/note="packaging signal for adenovirus serotype 5"
repeat_region 6274..6376
/note="ITR"
/note="inverted terminal repeat of human adenovirus
serotype 5"
rep_origin 6414..6794
/direction=RIGHT
/note="M13 ori"
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(6842..7636)
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
rep_origin 8468..9056
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatgga
61 tccatgcatg ttaattaaca tcatcaataa tataccttat tttggattga agccaatatg
121 ataatgaggg ggtggagttt gtgacgtggc gcggggcgtg ggaacggggc gggtgacgta
181 ggttttaggg cggagtaact tgtatgtgtt gggaattgta gttttcttaa aatgggaagt
241 gacgtaacgt gggaaaacgg aagtgacgat ttgaggaagt tgtgggtttt ttggctttcg
301 tttctgggcg taggttcgcg tgcggttttc tgggtgtttt ttgtggactt taaccgttac
361 gtcatttttt agtcctatat atactcgctc tgcacttggc ccttttttac actgtgactg
421 attgagctgg tgccgtgtcg agtggtgttt ttttaatagg ttttcttttt tactggtaag
481 gctgactgtt atggctgccg ctgtggaagc gctgtatgtt gttctggagc gggagggtgc
541 tattttgcct aggcaggagg gtttttcagg tgtttatgtg tttttctctc ctattaattt
601 tgttatacct cctatggggg ctgtaatgtt gtctctacgc ctgcgggtat gtattccccc
661 gggctatttc ggtcgctttt tagcactgac cgatgtgaat caacctgatg tgtttaccga
721 gtcttacatt atgactccgg acatgaccga ggagctgtcg gtggtgcttt ttaatcacgg
781 tgaccagttt ttttacggtc acgccggcat ggccgtagtc cgtcttatgc ttataagggt
841 tgtttttcct gttgtaagac aggcttctaa tgtttaaatg tttttttgtt attttatttt
901 gtgtttatgc agaaacccgc agacatgttt gagagaaaaa tggtgtcttt ttctgtggtg
961 gttccggagc ttacctgcct ttatctgcat gagcatgact acgatgtgct ttcttttttg
1021 cgcgaggctt tgcctgattt tttgagcagc accttgcatt ttatatcgcc gcccatgcaa
1081 caagctattg aattcgttta aactccctct caagtctgta tacggggaca cggacagcct
1141 tttcgtcacc gagcgtggac accggctcat ggaaaccaga ggtaagaaac gcatcaaaaa
1201 gcatggggga aacctggttt ttgaccccga acggccagag ctcacctggc tcgtggaatg
1261 cgagaccgtc tgcggggcct gcggcgcgga tgcctactcc ccggaatcgg tatttctcgc
1321 gcccaagctc tacgccctca aaagtctgca ctgcccctcg tgcggcgcct cctccaaggg
1381 caagctgcgc gccaagggcc acgccgcgga ggggctggac tatgacacca tggtcaaatg
1441 ctacctggcc gacgcgcagg gcgaagaccg gcagcgcttc agcaccagca ggaccagcct
1501 caagcgcacc ctggccagcg cgcagcccgg agcgcacccc ttcaccgtga cccagactac
1561 gctgacgagg accctgcgcc cgtggaaaga catgaccctg gcccgtctgg acgagcaccg
1621 actactgccg tacagcgaaa gccgccccaa cccgcgaaac gaggagatat gctggatcga
1681 gatgccgtag agcaggtgac cgagctgtgg gaccgcctgg aactgcttgg tcaaacgctc
1741 aaaagcatgc ctacggcgga cggtctcaaa ccgttgaaaa actttgcttc cttgcaagaa
1801 ctgctatcgc tgggcggcga gcgccttctg gcggatttgg tcagggaaaa catgcgagtc
1861 agggacatgc ttaacgaagt ggcccccctg ctcagggatg acggcagctg cagctctctt
1921 aactaccagt tgcagccggt aataggtgtg atttacgggc ccaccggctg cggtaagtcg
1981 cagctgctca ggaacctgct ttcttcccag ctgatctccc ctaccccgga aaccgttttc
2041 ttcatcgccc cgcaggtaga catgatcccc ccatctgaac tcaaagcgtg ggaaatgcaa
2101 atctgtgagg gtaactacgc ccctgggccg gatggaacca ttataccgca gtctggcacc
2161 ctccgcccgc gctttgtaaa aatggcctat gacgatctca tcctggaaca caactatgac
2221 gttagtgatc ccagaaatat cttcgcccag gccgccgccc gtgggcccat tgccatcatt
2281 atggacgaat gcatggaaaa tcttggaggt cacaagggcg tctccaagtt cttccacgca
2341 tttccttcta agctacatga caaatttccc aagtgcaccg gatacactgt gctggtggtt
2401 ctgcacaaca tgaatccccg gagggatatg gctgggaaca tagccaacct aaaaatacag
2461 tccaagatgc atctcatatc cccacgtatg cacccatccc agcttaaccg ctttgtaaac
2521 acttacacca agggcctgcc cctggcaatc agcttgctac tgaaagacat ttttaggcac
2581 cacgcccagc gctcctgcta cgactggatc atctacaaca ccaccccgca gcatgaagct
2641 ctgcagtggt gctacctcca ccccagagac gggcttatgc ccatgtatct gaacatccag
2701 agtcaccttt accacgtcct ggaaaaaata cacaggaccc tcaacgaccg agaccgctgg
2761 tcccgggcct accgcgcgcg caaaacccct aaataaagac agcaagacac ttgcttgatc
2821 caaatccaaa cagagtctgg ttttttattt atgttttaaa ccgcattggg aggggaggaa
2881 gccttcaggg cagaaacctg ctggcgcaga tccaacagct gctgagaaac gacattaagt
2941 tcccgggtca aagaatccaa ttgtgccaaa agagccgtca acttgtcatc gcgggcggat
3001 gaacgggaag ctgcactgct tgcaagcggg ctcaggaaag caaagtcagt cacaatcccg
3061 cgggcggtgg ctgcagcggc tgaagcggcg gcggaggctg cagtctccaa cggcgttcca
3121 gacacggtct cgtaggtcaa ggtagtagag tttgcgggca ggacggggcg accatcaatg
3181 ctggagccca tcacattctg acgcaccccg gcccatgggg gcatgcgcgt tgtcaaatat
3241 gagctcacaa tgcttccatc aaacgagttg gtgctcatgg cggcggcggc tgctgcaaaa
3301 cagatacaaa actacataag acccccacct tatatattct ttcccaccct taaccacgcc
3361 cagatcctct agcagtgata aacgtctaat agtaatcaat tacggggtca ttagttcata
3421 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
3481 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
3541 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
3601 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
3661 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
3721 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
3781 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
3841 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
3901 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
3961 gtcagatccg ctagagatct ggtaccgtcg acgcggccgc tcgagcctaa gcttctagat
4021 aagatatccg atccaccgga tctagataac tgatcataat cagccatacc acatttgtag
4081 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
4141 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
4201 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
4261 aactcatcaa tgtatcttaa cgcggaggtt tatcgacgat ctgctagtga ttaatagtaa
4321 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
4381 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
4441 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
4501 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
4561 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
4621 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
4681 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
4741 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
4801 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
4861 ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtc gccaccatgg
4921 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
4981 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
5041 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
5101 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
5161 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
5221 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
5281 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
5341 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
5401 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
5461 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
5521 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
5581 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtccggac
5641 tcagatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt
5701 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
5761 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
5821 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
5881 tcaatgtatc ttaacgcgat caagctagct tgctagactc gactgactat aataataaaa
5941 cgccaacttt gacccggaac gcggaaaaca cctgagaaaa acacctgggc gagtctccac
6001 gtaaacggtc aaagtccccg cggccctaga caaatattac gcgctatgag taacacaaaa
6061 ttattcagat ttcacttcct cttattcagt tttcccgcga aaatggccaa atcttactcg
6121 gttacgccca aatttactac aacatccgcc taaaaccgcg cgaaaattgt cacttcctgt
6181 gtacaccggc gcacaccaaa aacgtcactt ttgccacatc cgtcgcttac atgtgttccg
6241 ccacacttgc aacatcacac ttccgccaca ctactacgtc acccgccccg ttcccacgcc
6301 ccgcgccacg tcacaaactc caccccctca ttatcatatt ggcttcaatc caaaataagg
6361 tatattattg atgatgttaa ttaagaatta attcgatcct gaatggcgaa tggacgcgcc
6421 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
6481 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
6541 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
6601 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
6661 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
6721 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
6781 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat tttaacaaaa
6841 ttcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga
6901 taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac
6961 gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga
7021 atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca
7081 cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg
7141 cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag
7201 tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa
7261 gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt
7321 gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt
7381 cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc
7441 gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa
7501 ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt
7561 gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc
7621 catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcagag cttgatcccc
7681 tgcgccatca gatccttggc ggcaagaaag ccatccagtt tactttgcag ggcttcccaa
7741 ccttaccaga gggcgcccca gctggcaatt ccggttcgct tgctgtccat aaaaccgccc
7801 agtctagcta tcgccatgta agcccactgc aagctacctg ctttctcttt gcgcttgcgt
7861 tttcccttgt ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg
7921 gactggcttt ctacgtgaaa aggatctagg tgaagatcct ttttgataat ctcatggctg
7981 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
8041 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
8101 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
8161 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
8221 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
8281 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
8341 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
8401 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
8461 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
8521 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
8581 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
8641 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
8701 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
8761 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
8821 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
8881 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
8941 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
9001 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
9061 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
9121 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
9181 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaag
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