基本信息
| 启动子: | CMV promoter |
| 终止子: | WPRE |
| 质粒大小: | 8322bp |
| 原核抗性: | 氨苄青霉素Amp |
| 筛选标记: | 杀稻瘟菌素Blast |
| 克隆菌株: | 大肠杆菌DH5α |
| 培养条件: | 37℃，有氧，LB |
质粒简介
pLenti CMV GFP Blast (659-1)质粒是一个慢病毒过表达质粒，CMV启动子驱动EGFP绿色荧光蛋白的表达
质粒图谱
质粒序列
LOCUS Exported 8322 bp ds-DNA circular SYN 26-OCT-2017
DEFINITION synthetic circular DNA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8322)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..8322
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 27..131
/gene="bla"
/label=AmpR promoter
CDS 132..992
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 1163..1751
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
LTR 2448..2628
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 2672..2797
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 3290..3523
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 4012..4129
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 4152..4455
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 4456..4659
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
regulatory 4749..4758
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 4755..5474
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
misc_feature 5490..6078
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
promoter 6134..6464
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 6315..6450
/label=SV40 ori
/note="SV40 origin of replication"
promoter 6512..6559
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 6578..6976
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=Blast
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
LTR 7063..7296
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 7368..7489
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin 7508..7643
/label=SV40 ori
/note="SV40 origin of replication"
rep_origin 7865..8320
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
ORIGIN
1 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
61 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
121 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
241 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
301 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
361 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
421 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
481 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
541 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
661 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
721 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
901 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
961 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
1021 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
1081 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
1141 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
1201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
1261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
1321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
1381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
1441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
1501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
1561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
1621 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
1681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
1741 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
1801 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
1861 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
1921 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
1981 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
2041 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
2101 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
2161 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat
2221 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc
2281 cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg
2341 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc
2401 gcattgcaga gatattgtat ttaagtgcct agctcgatac aataaacggg tctctctggt
2461 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
2521 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
2581 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
2641 cagggacctg aaagcgaaag ggaaaccaga gctctctcga cgcaggactc ggcttgctga
2701 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
2761 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
2821 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
2881 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
2941 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
3001 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
3061 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
3121 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt
3181 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
3241 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
3301 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcctcaatg acgctgacgg
3361 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
3421 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
3481 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
3541 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
3601 tggaacagat tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
3661 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
3721 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
3781 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
3841 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
3901 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
3961 gagagacaga gacagatcca ttcgattagt gaacggatct cgacggttaa cttttaaaag
4021 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
4081 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaatttta tcgataagct
4141 tgggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
4201 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
4261 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
4321 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
4381 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
4441 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
4501 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
4561 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
4621 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc
4681 gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccgact ctagaggatc
4741 caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
4801 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
4861 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
4921 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
4981 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
5041 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
5101 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
5161 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
5221 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
5281 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
5341 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
5401 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
5461 agctgtacaa gtaaagcggc cgcgtcgaca atcaacctct ggattacaaa atttgtgaaa
5521 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
5581 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
5641 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
5701 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
5761 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
5821 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
5881 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
5941 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
6001 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
6061 tttgggccgc ctccccgcct ggaattctgc agatatccgg ttagtaatga gtttggaatt
6121 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc caggcaggca
6181 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
6241 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
6301 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
6361 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
6421 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
6481 gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat
6541 cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagcctt tgtctcaaga
6601 agaatccacc ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga
6661 ctacagcgtc gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt
6721 atatcatttt actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc
6781 ggcagctggc aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag
6841 cccctgcgga cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt
6901 gaaggacagt gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta
6961 tgtgtgggag ggctaagcac aattcgagct cggtaccttt aagaccaatg acttacaagg
7021 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctagctcact
7081 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
7141 gagcctggga gctctctggc tgcctaggga acccactgct taagcctcaa taaagcttgc
7201 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
7261 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
7321 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
7381 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
7441 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
7501 tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
7561 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
7621 gtgaggaggc ttttttggag gcctaggctt ttgcgtcgag acgtacccaa ttcgccctat
7681 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
7741 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
7801 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
7861 cgcgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
7921 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
7981 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
8041 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
8101 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
8161 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
8221 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
8281 tttaacgcga attttaacaa aatattaacg tttacaattt cc
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