基本信息
| 启动子: | SV40 |
| 终止子: | SV40 poly(A) signal |
| 质粒分类: | 哺乳细胞载体;蛋白过表达质粒 |
| 质粒大小: | 4976bp |
| 原核抗性: | 氨苄青霉素Amp |
| 筛选标记: | 甲氨蝶呤MTX |
| 克隆菌株: | 大肠杆菌DH5α |
| 培养条件: | 37℃，有氧，LB |
| 表达宿主: | 哺乳细胞 |
| 培养条件: | 37℃，有氧，LB |
| 5'测序引物: | SV40pro-F: TATTTATGCAGAGGCCGAGG |
| 3'测序引物: | SV40pA-R: GAAATTTGTGATGCTATTGC |
质粒简介
pSV2-DHFR载体含有的是小鼠的DHFR基因，同时该载体是哺乳动物细胞和大肠杆菌穿梭质粒，在DHFR细胞系中，可以通过甲氨蝶呤对转染的细胞进行筛选。质粒图谱
质粒序列
LOCUS Exported 4976 bp ds-DNA circular SYN 27-DEC-2018
DEFINITION synthetic circular DNA
KEYWORDS pSV2-DHFR
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4976)
AUTHORS Triple Threat
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..4976
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 10..339
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 190..325
/note="SV40 ori"
/note="SV40 origin of replication"
CDS 438..1001
/codon_start=1
/gene="Mus musculus Dhfr"
/product="mouse dihydrofolate reductase"
/note="DHFR"
/translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
PGVLSEVQEEKGIKYKFEVYEKKD"
intron 1158..1223
/note="small t intron"
/note="simian virus 40 (SV40) small t antigen intron"
CDS 1353..1373
/codon_start=1
/product="nuclear localization signal of SV40 large T
antigen"
/note="SV40 NLS"
/translation="PKKKRKV"
polyA_signal 1798..1932
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter 2785..2889
/gene="bla"
/note="AmpR promoter"
CDS 2890..3750
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 3921..4509
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 4695..4835
/note="bom"
/note="basis of mobility region from pBR322"
ORIGIN
1 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
61 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
121 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
181 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
241 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
301 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttggggg gggggacagc
361 tcagggctgc gatttcgcgc caaacttgac ggcaatccta gcgtgaaggc tggtaggatt
421 ttatccccgc tgccatcatg gttcgaccat tgaactgcat cgtcgccgtg tcccaaaata
481 tggggattgg caagaacgga gacctaccct ggcctccgct caggaacgag ttcaagtact
541 tccaaagaat gaccacaacc tcttcagtgg aaggtaaaca gaatctggtg attatgggta
601 ggaaaacctg gttctccatt cctgagaaga atcgaccttt aaaggacaga attaatatag
661 ttctcagtag agaactcaaa gaaccaccac gaggagctca ttttcttgcc aaaagtttgg
721 atgatgcctt aagacttatt gaacaaccgg aattggcaag taaagtagac atggtttgga
781 tagtcggagg cagttctgtt taccaggaag ccatgaatca accaggccac ctcagactct
841 ttgtgacaag gatcatgcag gaatttgaaa gtgacacgtt tttcccagaa attgatttgg
901 ggaaatataa acttctccca gaatacccag gcgtcctctc tgaggtccag gaggaaaaag
961 gcatcaagta taagtttgaa gtctacgaga agaaagacta acaggaagat gctttcaagt
1021 tctctgctcc cctcctaaag ctatgcattt ttataagacc atgggacttt tgctggcttt
1081 agatctttgt gaaggaacct tacttctgtg gtgtgacata attggacaaa ctacctacag
1141 agatttaaag ctctaaggta aatataaaat ttttaagtgt ataatgtgtt aaactactga
1201 ttctaattgt ttgtgtattt tagattccaa cctatggaac tgatgaatgg gagcagtggt
1261 ggaatgcctt taatgaggaa aacctgtttt gctcagaaga aatgccatct agtgatgatg
1321 aggctactgc tgactctcaa cattctactc ctccaaaaaa gaagagaaag gtagaagacc
1381 ccaaggactt tccttcagaa ttgctaagtt ttttgagtca tgctgtgttt agtaatagaa
1441 ctcttgcttg ctttgctatt tacaccacaa aggaaaaagc tgcactgcta tacaagaaaa
1501 ttatggaaaa atattctgta acctttataa gtaggcataa cagttataat cataacatac
1561 tgttttttct tactccacac aggcatagag tgtctgctat taataactat gctcaaaaat
1621 tgtgtacctt tagcttttta atttgtaaag gggttaataa ggaatatttg atgtatagtg
1681 ccttgactag agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
1741 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
1801 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
1861 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
1921 catgtctgga tccccaggaa gctcctctgt gtcctcataa accctaacct cctctacttg
1981 agaggacatt ccaatcatag gctgcccatc caccctctgt gtcctcctgt taattaggtc
2041 acttaacaaa aaggaaattg ggtaggggtt tttcacagac cgctttctaa gggtaatttt
2101 aaaatatctg ggaagtccct tccactgctg tgttccagaa gtgttggtaa acagcccaca
2161 aatgtcaaca gcagaaacat acaagctgtc agctttgcac aagggcccaa caccctgctc
2221 atcaagaagc actgtggttg ctgtgttagt aatgtgcaaa acaggaggca cattttcccc
2281 acctgtgtag gttccaaaat atctagtgtt ttcattttta cttggatcag gaacccagca
2341 ctccactgga taagcattat ccttatccaa aacagccttg tggtcagtgt tcatctgctg
2401 actgtcaact gtagcatttt ttggggttac agtttgagca ggatatttgg tcctgtagtt
2461 tgctaacaca ccctgcagct ccaaaggttc cccaccaaca gcaaaaaaat gaaaatttga
2521 cccttgaatg ggttttccag caccattttc atgagttttt tgtgtccctg aatgcaagtt
2581 taacatagca gttaccccaa taacctcagt tttaacagta acagcttccc acatcaaaat
2641 atttccacag gttaagtcct catttaaatt aggcaaagga attcttgaag acgaaagggc
2701 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
2761 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
2821 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
2881 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
2941 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
3001 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
3061 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
3121 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
3181 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
3241 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
3301 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
3361 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
3421 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
3481 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
3541 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
3601 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
3661 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
3721 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
3781 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
3841 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
3901 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
3961 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
4021 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
4081 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
4141 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
4201 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
4261 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
4321 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
4381 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
4441 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
4501 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
4561 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
4621 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
4681 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
4741 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
4801 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
4861 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
4921 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcag
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