| 质粒名称: | NPY-GFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 74629 |
| 存储实验室: | Justin Taraska |
|
载体骨架基本信息 |
|
| 载体名称: | GFP-N1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4700bp |
| 完整质粒大小: | 5016bp |
| 载体类型: | 哺乳细胞表达，Bacterial Expression |
| 筛选标记: | 新霉素（G418） |
| 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: | Neuropeptide Y、NPY |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 291bp |
| GenBank编号 | NM_000905 |
| 启动子: | CMV |
| 标签/融合蛋白: | mEGFP (C terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | HindIII (unknown if destroyed) |
| 3’克隆位点: | Acc65I (unknown if destroyed) |
| 5’测序引物 | CMV-F |
| 3’测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | NPY-GFP was a gift from Justin Taraska (Addgene plasmid # 74629 ; http://n2t.net/addgene:74629 ; RRID:Addgene_74629) |
买家导航
NPY-GFP载体质粒基本信息
NPY-GFP质粒图谱载体图谱和NPY-GFP载体序列质粒序列多克隆位点信息
NPY-GFP质粒载体简介
NPY-GFP质粒序列载体序列
LOCUS NPY-GFP 5016 bp DNA circular SYN 14-APR-2025
DEFINITION human neuropeptide Y fused to GFP.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5016)
TITLE NPY-GFP
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5016)
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"Unpublished"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: NPY-GFP
FEATURES Location/Qualifiers
source 1..5016
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 63..366
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 367..570
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 521..541
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 958..967
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS join(964..966,967..969,970..1683)
/codon_start=1
/product="enhanced GFP with monomerizing A206K mutation
(Zacharias et al., 2002)"
/label=mEGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1009..1030)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1270..1289)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1617..1638
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 1806..1927
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1843..1862)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1897..1916
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1934..2389)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2071..2092)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2283..2302
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2416..2520
/gene="bla"
/label=AmpR promoter
promoter 2522..2879
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
enhancer 2522..2713
/label=SV40 enhancer
/note="enhancer for the SV40 early promoter (Herr, 1993)"
primer_bind complement(2545..2565)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2730..2865
/label=SV40 ori
/note="SV40 origin of replication"
CDS 2914..3708
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
CDS 2914..3708
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2968..2987)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3578..3597
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3896..3915)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3940..3987
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4316..4904
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4805..4824
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
541 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc
601 taccggactc agatctcgag ctcaagctta gccaccatgc taggtaacaa gcgactgggg
661 ctgtccggac tgaccctcgc cctgtccctg ctcgtgtgcc tgggtgcgct ggccgaggcg
721 tacccctcca agccggacaa cccgggcgag gacgcaccag cggaggacat ggccagatac
781 tactcggcgc tgcgacacta catcaacctc atcaccaggc agagatatgg aaaacgatcc
841 agcccagaga cactgatttc agacctcttg atgagagaaa gcacagaaaa tgttcccaga
901 actcggcttg aagaccctgc aatgtggacg gtaccgcggg cccgggatcc accggtcgcc
961 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
1021 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
1081 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
1141 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
1201 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
1261 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
1321 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
1381 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
1441 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
1501 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
1561 cactacctga gcacccagtc caaactgagc aaagacccca acgagaagcg cgatcacatg
1621 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
1681 taaagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
1741 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
1801 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
1861 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
1921 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
1981 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
2041 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
2101 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
2161 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
2221 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
2281 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
2341 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
2401 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
2461 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
2521 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
2581 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
2641 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
2701 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
2761 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
2821 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
2881 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
2941 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
3001 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
3061 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
3121 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
3181 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
3241 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
3301 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
3361 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
3421 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
3481 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
3541 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
3601 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
3661 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
3721 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
3781 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
3841 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
3901 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
3961 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
4021 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
4081 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
4141 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
4201 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
4261 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
4321 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
4381 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
4441 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
4501 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
4561 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
4621 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
4681 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
4741 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
4801 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
4861 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
4921 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
4981 cccctgattc tgtggataac cgtattaccg ccatgc
//
