| 质粒名称: | pSL1180-HR-PUbECFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 47917 |
| 存储实验室: | Leslie Vosshall |
|
载体骨架基本信息 |
|
| 载体名称: | pSL1180 |
| 载体出品公司: | Clontech |
| 空载体大小: | 5066 bp |
| 完整质粒大小: | 5825 bp |
| 载体类型: | Insect Expression |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | Top10 |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Enhanced Cyan Fluorescent Protein |
| 物种来源: | Aequorea victoria |
| 插入大小: | 759 bp |
| 启动子: | Aedes aegypti polyubiquitin (PUb) |
| 标签/融合蛋白: | Aedes aegypti polyubiquitin (PUb) |
| 克隆方法: | Ligation Independent |
| 5’测序引物 | M13_reverse_primer |
| 3’测序引物 |
M13_forward20_primer
(Common Sequencing Primers)
|
|
文献引用方法 |
|
| 材料和方法部分: | pSL1180-HR-PUbECFP was a gift from Leslie Vosshall (Addgene plasmid # 47917 ; http://n2t.net/addgene:47917 ; RRID:Addgene_47917) |
买家导航
pSL1180-HR-PUbECFP载体质粒基本信息
pSL1180-HR-PUbECFP质粒图谱载体图谱和pSL1180-HR-PUbECFP载体序列质粒序列多克隆位点信息
pSL1180-HR-PUbECFP质粒载体简介
pSL1180-HR-PUbECFP质粒序列载体序列
LOCUS pSL1180-HR-PUbECFP 5825 bp DNA circular SYN 14-APR-2025
DEFINITION Donor plasmid for homologous recombination that expresses ECFP
consitutively using the Aedes aegypti polyubiquitin (PUb) promoter..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5825)
TITLE Vosshall lab Aedes aegypti HR donor plasmid
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5825)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT Sequence Label: pSL1180-HR-PUbECFP
FEATURES Location/Qualifiers
source 1..5825
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 106..127
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter join(142..147,148..165,166..172)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 180..196
/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)."
primer_bind 185..207
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 204..220
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 204..220
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
polyA_signal 289..423
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(313..332)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 367..386
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(564..584)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS complement(join(594..1304,1305..1307,1308..1310))
/codon_start=1
/product="enhanced CFP"
/label=ECFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(636..657)
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind 985..1004
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1244..1265
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2953..2970)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(2953..2969)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2962..2984)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 3182..3637
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3269..3288)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3479..3500
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3654..3673)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3773..3795
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(3833..3851)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3919..4023
/gene="bla"
/label=AmpR promoter
CDS join(4024..4092,4093..4884)
/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(4242..4261)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5055..5643
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5544..5563
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5797..5814
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg aattcgaatg
241 gccatgggac gtcgacctga ggtaattata acccgggccc tatatatgga tccagacatg
301 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
361 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
421 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
481 ttttaaagca agtaaaacct ctacaaatgt ggtatggctg attatgatca gttatctaga
541 tccggtggat cttacgggtc ctccaccttc cgctttttct tgggtcgaga tctcttgtac
601 agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga
661 tcgcgcttct cgttggggtc tttgctcagg gcggactggg tgctcaggta gtggttgtcg
721 ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc
781 acgctgccgt cctcgatgtt gtggcggatc ttgaagttgg ccttgatgcc gttcttctgc
841 ttgtcggcgg tgatatagac gttgtggctg atgtagttgt actccagctt gtgccccagg
901 atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg
961 ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg
1021 cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg
1081 tcggggtagc ggctgaagca ctgcacgccc caggtcaggg tggtcacgag ggtgggccag
1141 ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca
1201 tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg
1261 accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat gctagcgttg
1321 aaatctctgt tgagcagaaa aagaaacgag gaaacgcttg agtaattggt tgtgaaatgc
1381 aaactctcat ttgatattga ttcattgcct ttggcttcga gcacgacacg acaggtttta
1441 aacttgtttt gctttgtctg cgtttgcagt cgcaggccaa gtgaaaaata tacacttgaa
1501 ggtgatgacg tcacaacaac gccctacttt taagtgaaaa attaacttgt tttcgacttt
1561 gaactacgta gttttgaaat tgcgtatctt caagttttca cgatttcctc aaggtttttc
1621 tcgatatgtg ttaatattac cttaatgggt aattaccatc aaaatattta ttttagatat
1681 gtgacggagc aaatacgtta ttcttattat tctagaaatt taattcaatt agtagcgatg
1741 attcaacgaa atatgattat cgctgtgaat cacaattggg ttttatcaat gatgatgaaa
1801 ctgcgttgca aattttcact aatcactcaa agctcaatag tcgccatctt ggaaaatagt
1861 ttgctcattc aaagacaaag gaatcattca ccaaactagt tttcgctcat agctataatt
1921 tcatcaatta atttacctac cttcactaga agattccctt caccgaaatg cactttcacc
1981 aatttaatag taattgtcct ttgaataaag ctttgttcac tctgaaattt tctcctctgg
2041 ctaattggat cactcttttt cactagagac ttcacttcac ttgcactggc actgcttact
2101 tgggccgcgt aatgttcact ccactaggaa acgtattcga ttgagctggt ttcgcctttg
2161 caggggcgtt ttatagacac tgtcgtagtg gtggttgtac ttctagaaaa tttcagcaat
2221 acattcatac atatacctct gttcggttgg atggctctat atcgatcgaa tatgggtacc
2281 atccctgtgt ctgaatggac agcaaaacgt gcttgtgtct gttagtcgtt cattctgtac
2341 cttgaacgat gcagttcaac ttctggcaaa gacgtcaatg tacctaccct tcgtgtatat
2401 ggcatagaga ggaagatgtg cgaatgcctt tttcgataga gaaaggattt cctgatttga
2461 tcgacaattt cggtagctaa tcttagcttc gaatgtaatc tgataaccaa atccagagaa
2521 aaatcagtaa tttgggaatt tcacttgaat cattctaatt gcatcatttg ctgtattagt
2581 gcagtcagca agtgacgtca acccttctaa atcgatatac ttctgggaag ctttctttct
2641 tgtctggctc agctggtgcc aaggcaaatt ataattggat tcaatgcaca agctacatgt
2701 aaagatagct agcatgcgca aatttaaagc gctgatatca gcgctttaaa tttgcgcatg
2761 ctagctatag ttctagaggt accggttgtt aacgttagcc ggctacgtat actccggaat
2821 attaataggc ctaggatgca tatggcggcc gcctgcagct ggcgccatcg atacgcgtac
2881 gtcgcgaccg cggacatgta cagagctcga gaagtactag tggccacgtg ggccgtgcac
2941 cttaagcttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
3001 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
3061 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
3121 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
3181 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
3241 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
3301 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
3361 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
3421 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
3481 ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc ttttgattta
3541 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
3601 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
3661 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
3721 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
3781 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
3841 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
3901 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
3961 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
4021 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
4081 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
4141 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
4201 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
4261 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
4321 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
4381 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
4441 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
4501 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
4561 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
4621 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
4681 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
4741 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
4801 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
4861 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
4921 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
4981 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
5041 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
5101 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
5161 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
5221 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
5281 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
5341 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
5401 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
5461 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
5521 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
5581 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
5641 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
5701 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
5761 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
5821 gaaga
//
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