| 质粒名称: | sfGFP-C1 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 54579 |
| 存储实验室: | Michael Davidson, Geoffrey Waldo |
|
载体骨架基本信息 |
|
| 载体名称: | sfGFP-C1 |
| 空载体大小: | 4750bp |
| 完整质粒大小: | 4731 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | sfGFP (N terminal on backbone) |
| 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: | NO |
| 标签/融合蛋白: | sfGFP (N terminal on backbone) |
|
文献引用方法 |
|
| 材料和方法部分: | |
| 参考文献部分: | |
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 sfGFP-C1 4731 bp ds-DNA circular SYN 16-AUG-2021
DEFINITION Localization: C1 Cloning Vector, Excitation: 485, Emission: 507.
ACCESSION .
VERSION .
KEYWORDS sfGFP-C1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4731)
AUTHORS Pedelacq JD, Cabantous S, Tran T, Terwilliger TC, Waldo GS
TITLE Engineering and characterization of a superfolder green fluorescent
protein.
JOURNAL Nat Biotechnol. 2006 Jan . 24(1):79-88.
PUBMED 16369541
REFERENCE 2 (bases 1 to 4731)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 16, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..4731
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 91..394
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 395..598
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 549..569
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 637..646
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 643..1359
/codon_start=1
/product="GFP variant that folds robustly even when fused
to poorly folded proteins (Pedelacq et al., 2006)"
/label=superfolder GFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
EFVTAAGITHGMDELYK"
primer_bind complement(688..709)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind 1296..1317
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 1360..1425
/label=MCS
/note="multiple cloning site"
polyA_signal 1549..1670
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1586..1605)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1640..1659
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(1677..2132)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1814..1835)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2026..2045
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2159..2263
/gene="bla"
/label=AmpR promoter
promoter 2265..2622
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2288..2308)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 2473..2608
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2535..2554
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2657..3451
/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(2711..2730)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3321..3340
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(3639..3658)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 3683..3730
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4059..4647
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4548..4567
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc
61 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
121 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
181 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
241 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
301 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
361 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
421 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
481 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
541 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
601 tttagtgaac cgtcagatcc gctagcgcta ccggtcgcca ccatggtgag caagggcgag
661 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
721 aagttcagcg tgcgcggcga gggcgagggc gatgccacca acggcaagct gaccctgaag
781 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
841 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcgccacga cttcttcaag
901 tccgccatgc ccgaaggcta cgtccaggag cgcaccatca gcttcaagga cgacggcacc
961 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
1021 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacttc
1081 aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
1141 aagatccgcc acaacgtgga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
1201 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
1261 gtgctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
1321 gccgccggga tcactcacgg catggacgag ctgtacaagt ccggactcag atctcgagct
1381 caagcttcga attctgcagt cgacggtacc gcgggcccgg gatccaccgg atctagataa
1441 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
1501 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
1561 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
1621 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat
1681 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
1741 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
1801 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
1861 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
1921 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
1981 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
2041 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
2101 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
2161 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
2221 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
2281 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
2341 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
2401 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
2461 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
2521 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
2581 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
2641 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
2701 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
2761 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
2821 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
2881 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
2941 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
3001 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
3061 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
3121 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
3181 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
3241 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
3301 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
3361 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
3421 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
3481 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
3541 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
3601 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
3661 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
3721 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
3781 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
3841 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
3901 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
3961 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
4021 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
4081 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
4141 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
4201 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
4261 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
4321 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
4381 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
4441 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
4501 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
4561 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
4621 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
4681 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga t
//
