质粒名称: | AbVec1.1-IGLC2-XhoI |
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出品公司: | Addgene |
目录编号: | 99575 |
存储实验室: | Hedda Wardemann |
载体骨架基本信息 |
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载体名称: | pUC19 |
空载体大小: |
4976 bp
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载体类型: | 哺乳细胞表达 |
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 |
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启动子: | hCMV IE1 gene promoter |
克隆方法: | 限制性内切酶 |
5’测序引物 |
GCTTCGTTAGAACGCGGCTAC
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文献引用方法 |
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材料和方法部分: |
AbVec1.1-IGLC2-XhoI was a gift from Hedda Wardemann (Addgene plasmid # 99575 ; http://n2t.net/addgene:99575 ; RRID:Addgene_99575)
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参考文献部分: | Efficient generation of monoclonal antibodies from single human B cells by single cell RT-PCR and expression vector cloning. Tiller T, Meffre E, Yurasov S, Tsuiji M, Nussenzweig MC, Wardemann H. J Immunol Methods. 2008 Jan 1;329(1-2):112-24. Epub 2007 Oct 31. 10.1016/j.jim.2007.09.017 PubMed 17996249 |
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载体序列
LOCUS AbVec1.1-IGLC2-XhoI 4976 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of immunoglobulin light chains in mammalian cells, human lambda 2 constant region. ACCESSION . VERSION . KEYWORDS AbVec1.1-IGLC2-XhoI SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4976) AUTHORS Tiller T, Meffre E, Yurasov S, Tsuiji M, Nussenzweig MC, Wardemann H TITLE Efficient generation of monoclonal antibodies from single human B cells by single cell RT-PCR and expression vector cloning. JOURNAL J Immunol Methods. 2008 Jan 1;329(1-2):112-24. Epub 2007 Oct 31. PUBMED 17996249 REFERENCE 2 (bases 1 to 4976) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4976 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 106..127 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 142..172 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 180..196 /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)." primer_bind 185..207 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 204..220 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 204..220 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" enhancer 249..628 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 629..832 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 783..803 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 829..853 /label=LNCX /note="Human CMV promoter, forward primer" promoter 1063..1081 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 1063..1080 /label=SP6 /note="SP6 promoter, forward primer" primer_bind 1094..1113 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 1511..1520 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" polyA_signal 1524..1658 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1561..1580) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1615..1634 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" promoter 1727..2084 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(1750..1770) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 1935..2070 /label=SV40 ori /note="SV40 origin of replication" primer_bind 1997..2016 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(2104..2121) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2104..2120) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2113..2135) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2333..2788 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2420..2439) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2630..2651 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2805..2824) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2924..2946 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(2984..3002) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3070..3174 /gene="bla" /label=AmpR promoter CDS 3175..4035 /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 complement(3393..3412) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4206..4794 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4695..4714 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4948..4965 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat ccaactggca 61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct 121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat 181 tgtgagcgga taacaatttc acacaggaaa cagctatgac atgattacga attaattcga 241 gctcgcccga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt 301 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg 361 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc 421 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc 481 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg 541 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat 601 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg 661 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag 721 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt 781 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt 841 gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg 901 ggaccgatcc agcctccgcg gccgggaacg gtgcattgga acgcggattc cccgtgccaa 961 gagtgacgta agtaccgcct atagagtcta taggcccacc cccttggctt cgttagaacg 1021 cggctacaat taatacataa ccttatgtat catacacata cgatttaggt gacactatag 1081 aataacatcc actttgcctt tctctccaca ggtgtccact cccaggtcca actgcacctc 1141 ggttctatcg attgaattcc accatgggat ggtcatgtat catccttttt ctagtagcaa 1201 ctgcaaccgg ttgactaact agcctcgagt gaggagcttc aagccaacaa ggccacactg 1261 gtgtgtctca taagtgactt ctacccggga gccgtgacag tggcctggaa ggcagatagc 1321 agccccgtca aggcgggagt ggagaccacc acaccctcca aacaaagcaa caacaagtac 1381 gcggccagca gctacctgag cctgacgcct gagcagtgga agtcccacag aagctacagc 1441 tgccaggtca cgcatgaagg gagcaccgtg gagaagacag tggcccctac agaatgttca 1501 tagaagcttg gccgccatgg cccaacttgt ttattgcagc ttataatggt tacaaataaa 1561 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 1621 tgtccaaact catcaatgta tcttatcatg tctggatcgg gaattaattc ggcgcagcac 1681 catggcctga aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga 1741 accatctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca 1801 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct 1861 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc 1921 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg 1981 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc 2041 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctg ttaacagctt 2101 ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa 2161 tcgccttgca gcacatcccc ctttcgccag ttggcgtaat agcgaagagg cccgcaccga 2221 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct 2281 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc 2341 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt 2401 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc 2461 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta 2521 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc 2581 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg 2641 ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt 2701 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat 2761 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat 2821 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct 2881 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt 2941 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta 3001 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg 3061 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg 3121 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt 3181 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt 3241 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg 3301 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa 3361 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt 3421 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag 3481 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt 3541 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga 3601 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt 3661 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta 3721 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg 3781 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc 3841 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt 3901 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg 3961 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg 4021 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa 4081 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa 4141 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga 4201 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 4261 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact 4321 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac 4381 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg 4441 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 4501 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga 4561 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc 4621 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg 4681 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc 4741 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc 4801 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt 4861 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc 4921 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaaga //
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