| 质粒名称: | pBABE-puro |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 1764 |
| 存储实验室: | Hartmut Land, Jay Morgenstern, Bob Weinberg |
|
载体骨架基本信息 |
|
| 载体名称: | pBABE-puro |
| 空载体大小: | 5169 bp |
| 完整质粒大小: | 5086 bp |
| 载体类型: | 哺乳细胞表达, Retroviral |
| 筛选标记: | Puromycin |
| 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: | 无 |
| 克隆方法: | 限制性内切酶 |
| 5’测序引物 | BABE 5' |
| 3’测序引物 | BABE 3' |
|
文献引用方法 |
|
| 材料和方法部分: | pBABE-puro was a gift from Hartmut Land & Jay Morgenstern & Bob Weinberg (Addgene plasmid # 1764 ; http://n2t.net/addgene:1764 ; RRID:Addgene_1764) |
| 参考文献部分: | Advanced mammalian gene transfer: high titre retroviral vectors with multiple drug selection markers and a complementary helper-free packaging cell line. Morgenstern JP, Land H. Nucleic Acids Res. 1990 Jun 25;18(12):3587-96. PubMed 2194165 |
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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 pBABE-puro 5086 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Retroviral vector for cloning and expressing your gene of interest.
Puromycin selection..
ACCESSION .
VERSION .
KEYWORDS pBABE-puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5086)
AUTHORS Morgenstern JP, Land H
TITLE Advanced mammalian gene transfer: high titre retroviral vectors with
multiple drug selection markers and a complementary helper-free
packaging cell line.
JOURNAL Nucleic Acids Res. 1990 Jun 25;18(12):3587-96.
PUBMED 2194165
REFERENCE 2 (bases 1 to 5086)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5086
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 8..477
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 542..741
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 942..1358
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1288..1310
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1326..1342
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
primer_bind complement(1416..1436)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 1421..1750
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 1601..1736
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 1663..1682
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 1760..2359
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(1760..1779)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 2256..2276
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 2580..3049
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(3179..3196)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3350..3938)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3430..3449)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4098..4958)
/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 4721..4740
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4959..5063)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatccca gtgtggtggt
1381 acgtaggaat tcgccagcac agtggtcgac cctgtggaat gtgtgtcagt tagggtgtgg
1441 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
1501 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
1561 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
1621 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
1681 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
1741 cttttgcaaa aagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
1801 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
1861 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
1921 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
1981 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
2041 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
2101 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
2161 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
2221 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
2281 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
2341 ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc
2401 acgaccccat gcatcgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
2461 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
2521 gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc
2581 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
2641 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
2701 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga
2761 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
2821 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
2881 aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt
2941 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
3001 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac atgcagcatg
3061 tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca tagttgcatt
3121 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
3181 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
3241 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
3301 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
3361 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
3421 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
3481 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
3541 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
3601 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
3661 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
3721 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
3781 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
3841 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
3901 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
3961 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
4021 caaaaaggat cttcacctag atccttttgc ggccgcaaat caatctaaag tatatatgag
4081 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
4141 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
4201 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
4261 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
4321 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
4381 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
4441 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
4501 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
4561 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
4621 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
4681 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
4741 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
4801 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
4861 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
4921 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
4981 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
5041 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccac
//
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