| 质粒名称: | pcDNA3 GFP LIC cloning vector (6D) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 30127 |
| 存储实验室: | Scott Gradia |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 6145 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | GFP（主干网上的C终端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | LIC位点vKoz/GFP（克隆过程中被破坏） |
| 3' 克隆位点: | LIC位点vKoz/GFP（克隆过程中被破坏） |
| 5' 测序引物 | CMV F (5'cgcaaatgggcggtaggcgtg) |
| 3' 测序引物 | GFP reverse (5'cagctcgaccaggatgggc3') |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3 GFP LIC cloning vector (6D) was a gift from Scott Gradia (Addgene plasmid # 30127 ; http://n2t.net/addgene:30127 ; RRID:Addgene_30127) |
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
质粒图谱.png)
载体简介
载体序列
LOCUS pcDNA3 GFP LIC cloning vector (6D) 6144 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA3 GFP LIC cloning vector (6D)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6144)
TITLE 6-series mammalian LIC expression vectors
REFERENCE 2 (bases 1 to 6144)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6144
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 131..510
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 511..714
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 665..685
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 759..778
/label=T7
/note="T7 promoter, forward primer"
promoter 759..777
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 858..1574
/codon_start=1
/product="enhanced GFP with monomerizing A206K mutation
(Zacharias et al., 2002)"
/label=mEGFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(900..921)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind 1508..1529
/label=EGFP-C
/note="EGFP, forward primer"
promoter complement(1593..1611)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1594..1611)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(1631..1648)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1637..1861
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1907..2335
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1994..2013)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2204..2225
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2344..2364)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2349..2678
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2529..2664
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2591..2610
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2745..3539
/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
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2799..2818)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3409..3428
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3713..3834
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3750..3769)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3804..3823
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3883..3899)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3883..3899)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3896..3918)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3907..3923
/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(3931..3961)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3976..3997
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4114..4131)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4285..4873)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4365..4384)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5044..5904)
/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 5667..5686
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5905..6009)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6084..6103)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
61 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
121 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
181 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
241 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
301 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
361 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
421 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
481 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
541 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
601 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
661 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
721 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
781 acccaagctt ggtaccgagc tcggatccct acttccaatc caatattggt agtgggagca
841 acggcagcag cggatccgtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
901 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgcgcggc gagggcgagg
961 gcgatgccac caacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
1021 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
1081 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
1141 agcgcaccat ctccttcaag gacgacggca cctacaagac ccgcgccgag gtgaagttcg
1201 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
1261 acatcctggg gcacaagctg gagtacaact tcaacagcca caacgtctat atcacggccg
1321 acaagcagaa gaacggcatc aaggcgaact tcaagatccg ccacaacgtc gaggacggca
1381 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
1441 tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc
1501 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
1561 agctgtacaa gtagtaatct agagggccct attctatagt gtcacctaaa tgctagagct
1621 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
1681 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
1741 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
1801 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
1861 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
1921 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
1981 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
2041 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
2101 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
2161 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
2221 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
2281 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
2341 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
2401 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
2461 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
2521 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
2581 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
2641 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
2701 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
2761 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
2821 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
2881 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
2941 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
3001 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
3061 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
3121 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
3181 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
3241 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
3301 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
3361 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
3421 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
3481 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
3541 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
3601 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
3661 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
3721 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
3781 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
3841 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
3901 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
3961 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
4021 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
4081 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
4141 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
4201 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
4261 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
4321 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
4381 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
4441 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
4501 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
4561 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
4621 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
4681 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
4741 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
4801 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
4861 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
4921 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
4981 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
5041 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
5101 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
5161 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
5221 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
5281 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
5341 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
5401 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
5461 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
5521 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
5581 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
5641 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
5701 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
5761 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
5821 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
5881 gacacggaaa tgttgaatac tcatactctt cctttttcat tattattgaa gcatttatca
5941 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
6001 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
6061 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
6121 ctgctccctg cttgtgtgtt ggag
//
