| 质粒名称: | pT4-CMV-GFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 117046 |
| 存储实验室: | Wolfgang Uckert |
|
载体骨架基本信息 |
|
| 载体名称: | pBluescript |
| 载体出品公司: | Stratagene |
| 空载体大小: | 3457 bp |
| 完整质粒大小: | 4999 bp |
| 载体修饰: | 插入的睡美人终端反向重复序列吞噬CMV-GFP转基因盒。 |
| 载体类型: | Transposon |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | GFP |
| 物种来源: | -- |
| 插入大小: | 720 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | CMV |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NotI(未破坏) |
| 3’克隆位点: | EcoRI (未破坏) |
| 5’测序引物 | GFP-FWD |
| 3’测序引物 | GFP-REV |
|
文献引用方法 |
|
| 材料和方法部分: | pT4-CMV-GFP was a gift from Wolfgang Uckert (Addgene plasmid # 117046 ; http://n2t.net/addgene:117046 ; RRID:Addgene_117046) |
买家导航
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 pT4-CMV-GFP 5036 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Optimized Sleeping Beauty Transposon Vector for GFP expression by a
CMV promoter.
ACCESSION .
VERSION .
KEYWORDS pT4-CMV-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5036)
TITLE Optimized Sleeping Beauty Transposon Vector with CMV-GFP cassette
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5036)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5036
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
intron 676..808
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
regulatory 813..822
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 819..1538
/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 819..1535
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(864..885)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1125..1144)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1472..1493
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 1545..1593
/note="synthetic polyadenylation signal"
primer_bind complement(1929..1949)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(1929..1947)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(1968..1984)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1968..1984)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1981..2003)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1992..2008
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(2016..2046)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2061..2082
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2199..2216)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2370..2958)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2450..2469)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3129..3989)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAAKLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 3752..3771
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3990..4094)
/gene="bla"
/label=AmpR promoter
rep_origin 4121..4576
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4208..4227)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4418..4439
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4702..4724
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 4716..4733
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 4717..4733
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 4743..4762
/label=T7
/note="T7 promoter, forward primer"
promoter 4743..4761
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag aggtcgttta gtgaaccgtc
601 agatcactag tagctttatt gcggtagttt atcacagtta aattgctaac gcagtcagtg
661 ctcgactgat cacaggtaag tatcaaggtt acaagacagg tttaaggagg ccaatagaaa
721 ctgggcttgt cgagacagag aagattcttg cgtttctgat aggcacctat tggtcttact
781 gacatccact ttgcctttct ctccacaggg cggccgccat ggtgagcaag ggcgaggagc
841 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
901 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
961 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
1021 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
1081 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
1141 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
1201 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
1261 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
1321 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
1381 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
1441 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
1501 ccgggatcac tctcggcatg gacgagctgt acaagtaaga attcaataaa atatctttat
1561 tttcattaca tctgtgtgtt ggttttttgt gtgtaattcg agcatcttac cgccatttat
1621 tcccagatct ttaaacaatt taaaggcaat gctaccaaat actaagcgcg tgtatgtaca
1681 cttctgaccc actgggaatg tgatgaaaga aataaaagct gaaatgaatc attctctcta
1741 ctattattct gatatttcac attcttaaaa taaagtggtg atcctaactg accttaagac
1801 agggaatctt tactcggatt aaatgtcagg aattgtgaaa aagtgagttt aaatgtattt
1861 ggctaaggtg tatgtaaact tccgacttca actgtagttt aaacgggccc ggtacccagc
1921 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
1981 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
2041 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
2101 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
2161 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
2221 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
2281 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
2341 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
2401 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
2461 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
2521 tacctgtccg cctttctccc ttcgggaacc gtggcgcttt ctcatagctc acgctgtagg
2581 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
2641 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
2701 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
2761 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
2821 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
2881 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
2941 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
3001 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
3061 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
3121 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
3181 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
3241 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
3301 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
3361 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
3421 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
3481 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
3541 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaactt ggccgcagtg
3601 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
3661 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
3721 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
3781 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
3841 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
3901 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
3961 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
4021 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
4081 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
4141 ttaagcgcgg cgggagtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
4201 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
4261 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
4321 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
4381 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
4441 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
4501 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
4561 ttaacgctta caatttccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
4621 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
4681 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg
4741 cgtaatacga ctcactatag ggcgaattgg agctagcgtt taaacctaca gttgaagtcg
4801 gaagtttaca tacacttaag ttggagtcat taaaactcgt ttttcaacta ctccacaaat
4861 ttcttgttaa caaacaatag ttttggcaag tcagttagga catctacttt gtgcatgaca
4921 caagtcattt ttccaacaat tgtttacaga cagattattt cacttataat tcactgtatc
4981 acaattccag tgggtcagaa gtgtacatac acgcgcttga ctgtgccttt aagctt
//
