| 质粒名称: | pT2/SVNeo |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 26553 |
| 存储实验室: | Perry Hackett |
|
载体骨架基本信息 |
|
| 载体名称: | pUC19 |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄西林 |
| 空载体大小: | 3712 bp |
| 载体类型: | 细菌表达 |
| 筛选标记: | 新霉素（用G418选择） |
| 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: | pT2/SVNeo |
| 插入大小: | 4928bp |
| 基因突变: | 增加了RMUT（80、87、D101） |
| 克隆方法: | 限制性酶 |
| 5' 克隆位点: | Ecori（未销毁） |
| 3' 克隆位点: | 八木（未销毁） |
| 5' 测序引物 | M13（-20）前进 |
| 3' 测序引物 | M13 Reverse |
|
文献引用方法 |
|
| 材料和方法部分: | pT2/SVNeo was a gift from Perry Hackett (Addgene plasmid # 26553 ; http://n2t.net/addgene:26553 ; RRID:Addgene_26553) |
| 参考文献部分: | Structure-function analysis of the inverted terminal repeats of the sleeping beauty transposon. Cui Z, Geurts AM, Liu G, Kaufman CD, Hackett PB. J Mol Biol. 2002 May 17. 318(5):1221-35. 10.1016/S0022-2836(02)00237-1 PubMed 12083513 |
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 pT2/SVNeo 5153 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pT2/SVNeo
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5153)
AUTHORS Cui Z, Geurts AM, Liu G, Kaufman CD, Hackett PB
TITLE Structure-function analysis of the inverted terminal repeats of the
sleeping beauty transposon.
JOURNAL J Mol Biol. 2002 May 17. 318(5):1221-35.
PUBMED 12083513
REFERENCE 2 (bases 1 to 5153)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5153
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 34..56
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 48..65
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 49..65
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 75..94
/label=T7
/note="T7 promoter, forward primer"
promoter 75..93
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(488..508)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 493..822
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 673..808
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 735..754
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 889..1683
/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(943..962)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 1553..1572
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 1857..1991
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1894..1913)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1948..1967
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(2414..2434)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(2414..2432)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(2453..2469)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2453..2469)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2466..2488)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2477..2493
/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(2501..2531)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2546..2567
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2684..2701)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2855..3443)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2935..2954)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3614..4474)
/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 4237..4256
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4475..4579)
/gene="bla"
/label=AmpR promoter
rep_origin 4606..5061
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4693..4712)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4903..4924
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
61 ccagtgagcg cgcgtaatac gactcactat agggcgaatt ggagctcgga tccctataca
121 gttgaagtcg gaagtttaca tacacttaag ttggagtcat taaaactcgt ttttcaacta
181 ctccacaaat ttcttgttaa caaacaatag ttttggcaag tcagttagga catctacttt
241 gtgcatgaca caagtcattt ttccaacaat tgtttacaga cagattattt cacttataat
301 tcactgtatc acaattccag tgggtcagaa gtttacatac actaagttga ctgtgccttt
361 aaacagcttg gaaaattcca gaaaatgatg tcatggcttt agaagcttcc cacaataagt
421 tgggtgaatt ttggctcatt cctcctttct ataggattga ggtcagagct ttgtgatgga
481 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
541 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
601 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
661 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
721 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
781 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgg
841 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
901 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
961 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
1021 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
1081 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
1141 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
1201 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
1261 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
1321 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
1381 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
1441 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
1501 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
1561 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
1621 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
1681 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
1741 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
1801 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
1861 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
1921 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
1981 atgtctggat cccttgaaat acatccacag gtacacctcc aattgactca aatgatgtca
2041 attagtctat cagaagcttg tggaaggcta ctcgaaatgt ttgacccaag ttaaacaatt
2101 taaaggcaat gctaccaaat actaattgag tgtatgtaaa cttctgaccc actgggaatg
2161 tgatgaaaga aataaaagct gaaatgaatc attctctcta ctattattct gatatttcac
2221 attcttaaaa taaagtggtg atcctaactg acctaagaca gggaattttt actaggatta
2281 aatgtcagga attgtgaaaa agtgagttta aatgtatttg gctaaggtgt atgtaaactt
2341 ccgacttcaa ctgtataggg atcctctagc tagagtcgac ctcgaggggg ggcccggtac
2401 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
2461 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
2521 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
2581 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
2641 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
2701 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
2761 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
2821 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
2881 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
2941 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
3001 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
3061 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
3121 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
3181 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
3241 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
3301 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
3361 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
3421 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
3481 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
3541 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
3601 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
3661 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
3721 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
3781 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
3841 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
3901 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
3961 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
4021 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
4081 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
4141 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
4201 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
4261 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
4321 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
4381 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
4441 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
4501 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
4561 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg
4621 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
4681 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
4741 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
4801 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
4861 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
4921 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
4981 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
5041 aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg
5101 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctg
//
