pT3-neo睡美人转座子表达载体-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心
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- 货 号:pT3-neo plasmid
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pT3-neo睡美人转座子表达载体
描述:
Sleeping Beauty (SB) transposon with G418 resistance
带有U3 MSCV启动子+Neomycin-G418真核筛选标记,IR-DR反向重复序列,Amp抗性。
Map图谱:

- Vector backbonepT3(Search Vector Database)
Backbone size w/o insert (bp)4222
Total vector size (bp)5017
- Vector typeMammalian Expression ; Sleeping Beauty transposon
- Selectable markersNeomycin (select with G418)
GROWTH IN BACTERIA
- Bacterial Resistance(s)Ampicillin
- Growth Temperature37°C
- Growth Strain(s)DH5alpha
- Copy numberHigh Copy
GENE/INSERT
- Gene/Insert nameNeo (codon optimized for human)
- SpeciesH. sapiens (human)
PromoterU3MSCV
CLONING INFORMATION
Cloning methodRestriction Enzyme
5′ cloning siteAgeI (not destroyed)
3′ cloning siteNotI (not destroyed)
5′ sequencing primerACTAACCAATCAGTTCGCTTCTC
3′ sequencing primerTAGAAGGCACAGTCGAGG
LOCUS Exported 5017 bp ds-DNA circular SYN 10-AGO-2015
DEFINITION .
ACCESSION .
VERSION .
KEYWORDS pT3-Neo
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5017)
AUTHORS Marco
TITLE Direct Submission
JOURNAL http://www.biovector.net
FEATURES Location/Qualifiers
source 1..5017
/organism="recombinant plasmid"
/mol_type="other DNA"
misc_feature 6..359
/note="IR/DR"
primer_bind complement(404..420)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 428..444
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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(452..482)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 497..518
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(806..1394)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1565..2425)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(2426..2530)
/gene="bla"
/note="AmpR promoter"
primer_bind 3004..3020
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 3040..3393
/note="IR/DR"
promoter 3518..3903
/note="U3 MSCV"
CDS 3914..4708
/codon_start=1
/note="Neo"
/translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
promoter complement(4743..4761)
/note="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
polyA_signal 4787..5011
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
ORIGIN
1 gatccccttg aaatacatcc acaggtacac ctccaattga ctcaaatgat gtcaattagt
61 ctatcagaag cttctaaagc catgacatca ttttctggaa ttttccaagc tgtttaaagg
121 cacagtcaac ttagtgtatg taaacttctg acccactgga attgtgatac agtgaattat
181 aagtgaaata atctgtctgt aaacaattgt tggaaaaatg acttgtgtca tgcacaaagt
241 agatgtccta actgacttgc caaaactatt gtttgttaac aagaaatttg tggagtagtt
301 gaaaaacgag ttttaatgac tccaacttaa gtgtatgtaa acttccgact tcaactgtat
361 atctagagtc gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg
421 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
481 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
541 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
601 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
661 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
721 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
781 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
841 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
901 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
961 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
1021 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
1081 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
1141 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
1201 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
1261 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
1321 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
1381 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
1441 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
1501 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
1561 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
1621 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
1681 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
1741 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
1801 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
1861 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
1921 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
1981 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
2041 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
2101 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
2161 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
2221 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
2281 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
2341 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
2401 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
2461 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
2521 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
2581 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
2641 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
2701 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
2761 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
2821 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
2881 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
2941 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
3001 gttgtaaaac gacggccagt gaattcgagc tcggtaccct acagttgaag tcggaagttt
3061 acatacactt aagttggagt cattaaaact cgtttttcaa ctactccaca aatttcttgt
3121 taacaaacaa tagttttggc aagtcagtta ggacatctac tttgtgcatg acacaagtca
3181 tttttccaac aattgtttac agacagatta tttcacttat aattcactgt atcacaattc
3241 cagtgggtca gaagtttaca tacactaagt tgactgtgcc tttaaacagc ttggaaaatt
3301 ccagaaaatg atgtcatggc tttagaagct tctgatagac taattgacat catttgagtc
3361 aattggaggt gtacctgtgg atgtatttca agggaattct gtggaatgtg tgtcagttag
3421 ggtgtggaaa gtccccaggc tccccaggca ggcagaagta tgcaaagcat gcatatcgat
3481 actagtttaa ttaagatctc gagcacgaga ctagccgtcg agggaatgaa agaccccacc
3541 tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac
3601 tgagaataga gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa
3661 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca
3721 gatgcggtcc cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa
3781 ggacctgaaa atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct
3841 gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg
3901 ccgatccacc ggtatggccg aacaggatgg actgcacgca ggctcccccg ccgcttgggt
3961 ggaaagactg tttggatacg attgggctca gcagactatt ggatgctctg acgcagccgt
4021 gttccggctg agtgctcagg ggcgccccgt gctgtttgtc aagacagacc tgtcaggagc
4081 actgaacgag ctgcaggatg aagcagcacg actgagctgg ctggcaacca caggagtgcc
4141 ttgtgctgca gtcctggacg tggtcactga ggcaggacga gattggctgc tgctgggaga
4201 agtgccagga caggacctgc tgagctccca cctggcacca gctgagaagg tctccatcat
4261 ggcagatgcc atgcggagac tgcatactct ggaccccgcc acctgccctt tcgatcacca
4321 ggctaaacat cgcattgaga gggctcgcac ccgaatggaa gcaggcctgg tggaccagga
4381 cgatctggat gaggaacacc aggggctggc tcctgcagag ctgtttgcac ggctgaaagc
4441 cagaatgcca gacggagaag atctggtggt cacccatggc gacgcttgcc tgcccaacat
4501 catggtggag aatggcagat tcagtgggtt tattgattgt ggacgactgg gcgtcgccga
4561 cagataccag gatatcgccc tggctacaag agacattgca gaggaactgg gcggggaatg
4621 ggccgatagg ttcctggtgc tgtatggcat cgctgcccct gattcacaga ggattgcctt
4681 ttacagactg ctggatgagt tcttttgagc ggccgctcga gcatgcatct agagggccct
4741 attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt gccttctagt
4801 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
4861 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
4921 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
4981 aggcatgctg gggatgcggt gggctctatg gcttctg
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