| 质粒名称: | pCLucf |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 37328 |
| 存储实验室: | John Schiller |
|
载体骨架基本信息 |
|
| 载体名称: | custom |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Luciferase |
| 物种来源: | D. melanogaster (fly) |
| GenBank编号 | luciferase (a.k.a. C25G6.3) |
| 启动子: | CMV |
| 插入基因2: | GFP |
| 物种来源: |
Aequorea victoria
|
| 启动子: | SV40 |
|
文献引用方法 |
|
| 材料和方法部分: | pCLucf was a gift from John Schiller (Addgene plasmid # 37328 ; http://n2t.net/addgene:37328 ; RRID:Addgene_37328) |
| 参考文献部分: | Role of heparan sulfate in attachment to and infection of the murine female genital tract by human papillomavirus. Johnson KM, Kines RC, Roberts JN, Lowy DR, Schiller JT, Day PM. J Virol. 2009 Mar;83(5):2067-74. Epub 2008 Dec 10. 10.1128/JVI.02190-08 PubMed 19073722 |
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载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pCLucf 7053 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCLucf
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7053)
AUTHORS Johnson KM, Kines RC, Roberts JN, Lowy DR, Schiller JT, Day PM
TITLE Role of heparan sulfate in attachment to and infection of the murine
female genital tract by human papillomavirus.
JOURNAL J Virol. 2009 Mar;83(5):2067-74. Epub 2008 Dec 10.
PUBMED 19073722
REFERENCE 2 (bases 1 to 7053)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7053
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 138..517
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 518..729
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 680..700
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 726..750
/label=LNCX
/note="Human CMV promoter, forward primer"
intron 890..1022
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
primer_bind 1005..1024
/label=pMT2-F
/note="Synthetic intron, forward primer"
promoter 1067..1085
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 1135..1144
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1141..2793
/codon_start=1
/product="firefly luciferase"
/label=luciferase
/note="synthetic luc2 version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
promoter complement(2826..2843)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
polyA_signal 2861..2982
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2872..2891)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2926..2945
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
rep_origin 3168..3623
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3255..3274)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3465..3486
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 3740..4097
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3763..3783)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3948..4083
/label=SV40 ori
/note="SV40 origin of replication"
regulatory 4122..4131
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 4128..4847
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 4128..4844
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(4173..4194)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4434..4453)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4781..4802
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 4902..4950
/note="synthetic polyadenylation signal"
primer_bind complement(4991..5010)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5110..5132
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5170..5188)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5256..5360
/gene="bla"
/label=AmpR promoter
CDS 5361..6221
/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"
primer_bind complement(5579..5598)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6392..6980
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6881..6900
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
1081 ataggctagc ctcgagaatt ctctagagct tggcaatccg gtactgttgg taaagccacc
1141 atggaagatg ccaaaaacat taagaagggc ccagcgccat tctacccact cgaagacggg
1201 accgccggcg agcagctgca caaagccatg aagcgctacg ccctggtgcc cggcaccatc
1261 gcctttaccg acgcacatat cgaggtggac attacctacg ccgagtactt cgagatgagc
1321 gttcggctgg cagaagctat gaagcgctat gggctgaata caaaccatcg gatcgtggtg
1381 tgcagcgaga atagcttgca gttcttcatg cccgtgttgg gtgccctgtt catcggtgtg
1441 gctgtggccc cagctaacga catctacaac gagcgcgagc tgctgaacag catgggcatc
1501 agccagccca ccgtcgtatt cgtgagcaag aaagggctgc aaaagatcct caacgtgcaa
1561 aagaagctac cgatcataca aaagatcatc atcatggata gcaagaccga ctaccagggc
1621 ttccaaagca tgtacacctt cgtgacttcc catttgccac ccggcttcaa cgagtacgac
1681 ttcgtgcccg agagcttcga ccgggacaaa accatcgccc tgatcatgaa cagtagtggc
1741 agtaccggat tgcccaaggg cgtagcccta ccgcaccgca ccgcttgtgt ccgattcagt
1801 catgcccgcg accccatctt cggcaaccag atcatccccg acaccgctat cctcagcgtg
1861 gtgccatttc accacggctt cggcatgttc accacgctgg gctacttgat ctgcggcttt
1921 cgggtcgtgc tcatgtaccg cttcgaggag gagctattct tgcgcagctt gcaagactat
1981 aagattcaat ctgccctgct ggtgcccaca ctatttagct tcttcgctaa gagcactctc
2041 atcgacaagt acgacctaag caacttgcac gagatcgcca gcggcggggc gccgctcagc
2101 aaggaggtag gtgaggccgt ggccaaacgc ttccacctac caggcatccg ccagggctac
2161 ggcctgacag aaacaaccag cgccattctg atcacccccg aaggggacga caagcctggc
2221 gcagtaggca aggtggtgcc cttcttcgag gctaaggtgg tggacttgga caccggtaag
2281 acactgggtg tgaaccagcg cggcgagctg tgcgtccgtg gccccatgat catgagcggc
2341 tacgttaaca accccgaggc tacaaacgct ctcatcgaca aggacggctg gctgcacagc
2401 ggcgacatcg cctactggga cgaggacgag cacttcttca tcgtggaccg gctgaagagc
2461 ctgatcaaat acaagggcta ccaggtagcc ccagccgaac tggagagcat cctgctgcaa
2521 caccccaaca tcttcgacgc cggggtcgcc ggcctgcccg acgacgatgc cggcgagctg
2581 cccgccgcag tcgtcgtgct ggaacacggt aaaaccatga ccgagaagga gatcgtggac
2641 tatgtggcca gccaggttac aaccgccaag aagctgcgcg gtggtgttgt gttcgtggac
2701 gaggtgccta aaggactgac cggcaagttg gacgcccgca agatccgcga gattctcatt
2761 aaggccaaga agggcggcaa gatcgccgtg taataattct agtggatccc ccgggcggcc
2821 gcttcccttt agtgagggtt aatgcttcga gcagacatga taagatacat tgatgagttt
2881 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
2941 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
3001 cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcaa gtaaaacctc
3061 tacaaatgtg gtaaaatccg ataaggatcg atccgggctg gcgtaatagc gaagaggccc
3121 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag
3181 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
3241 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
3301 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
3361 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
3421 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
3481 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
3541 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
3601 caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
3661 atttcacacc gcatacgcgg atctgcgcag caccatggcc tgaaataacc tctgaaagag
3721 gaacttggtt aggtaccttc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
3781 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
3841 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
3901 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
3961 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatacag
4021 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
4081 gcctaggctt ttgcaaaaag cttgattggg atccaccggt cgccaccatg gtgagcaagg
4141 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
4201 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
4261 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
4321 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
4381 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
4441 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
4501 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
4561 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
4621 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
4681 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
4741 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
4801 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc ggcccgaaat
4861 gaccgaccaa gcgacgccca acctgccatc acgatggccg caataaaata tctttatttt
4921 cattacatct gtgtgttggt tttttgtgtg aatcgatagc gataaggatc cgcgtatggt
4981 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
5041 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
5101 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
5161 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
5221 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
5281 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
5341 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
5401 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
5461 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
5521 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
5581 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
5641 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
5701 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
5761 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
5821 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
5881 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
5941 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
6001 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
6061 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
6121 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
6181 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
6241 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
6301 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
6361 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
6421 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
6481 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
6541 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
6601 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
6661 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
6721 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
6781 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
6841 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
6901 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
6961 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
7021 gctggccttt tgctcacatg gctcgacaga tct
//
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