质粒名称: | AbVec2.0-IGHG1 |
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出品公司: | Addgene |
目录编号: | 80795 |
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: | 5741 bp |
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 |
文献引用方法 |
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材料和方法部分: | AbVec2.0-IGHG1 was a gift from Hedda Wardemann (Addgene plasmid # 80795 ; http://n2t.net/addgene:80795 ; RRID:Addgene_80795) |
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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AbVec2.0IGHG1载体基本信息
AbVec2.0IGHG1载体图谱质粒图谱和多克隆位点信息
AbVec2.0IGHG1载体简介
AbVec2.0IGHG1载体序列
LOCUS AbVec2.0-IGHG1 5741 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of secretory immunoglobulin heavy chains in mammalian cells, human IgG1 isotype. ACCESSION . VERSION . KEYWORDS AbVec2.0-IGHG1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5741) 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 5741) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5741 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 164..181 /label=L4440 /note="L4440 vector, forward primer" protein_bind 298..319 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 334..364 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 372..388 /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 377..399 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 396..412 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 396..412 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" enhancer 441..820 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 821..1024 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 975..995 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 1021..1045 /label=LNCX /note="Human CMV promoter, forward primer" promoter 1255..1273 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 1255..1272 /label=SP6 /note="SP6 promoter, forward primer" primer_bind 1286..1305 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 2468..2477 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" polyA_signal 2481..2615 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2518..2537) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2572..2591 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" promoter 2684..3041 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2707..2727) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2892..3027 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2954..2973 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(3061..3078) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3061..3077) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3070..3092) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3290..3745 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3377..3396) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3587..3608 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3762..3781) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3881..3903 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3941..3959) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4027..4131 /gene="bla" /label=AmpR promoter CDS 4132..4992 /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(4350..4369) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5163..10 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5652..5671 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt 61 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt 121 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga 181 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta 241 atccaactgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa 301 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat 361 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac 421 gaattaattc gagctcgccc gacattgatt attgactagt tattaatagt aatcaattac 481 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg 541 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc 601 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac 661 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa 721 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac 781 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta 841 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga 901 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa 961 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag 1021 agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca 1081 tagaagacac cgggaccgat ccagcctccg cggccgggaa cggtgcattg gaacgcggat 1141 tccccgtgcc aagagtgacg taagtaccgc ctatagagtc tataggccca cccccttggc 1201 ttcgttagaa cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag 1261 gtgacactat agaataacat ccactttgcc tttctctcca caggtgtcca ctcccaggtc 1321 caactgcacc tcggttctat cgattgaatt ccaccatggg atggtcatgt atcatccttt 1381 ttctagtagc aactgcaacc ggttgactaa ctagctggtc gaccaagggc ccaagcgtct 1441 tccccctggc accctcctcc aagagcacct ctgggggcac agcggccctg ggctgcctgg 1501 tcaaggacta cttccccgag ccggtgacgg tgtcgtggaa ctcaggcgcc ctgaccagcg 1561 gcgtgcacac cttcccggct gtcctacagt cctcaggact ctactccctc agcagcgtgg 1621 tgaccgtgcc ctccagcagc ttgggcaccc agacctacat ctgcaacgtg aatcacaagc 1681 ccagcaacac caaggtggac aagagagttg agcccaaatc ttgtgacaaa actcacacat 1741 gcccaccgtg cccagcacct gaactcctgg ggggaccgtc agtcttcctc ttccccccaa 1801 aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg gtggtggacg 1861 tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg gaggtgcata 1921 atgccaagac aaagccgcgg gaggagcagt acaacagcac gtaccgtgtg gtcagcgtcc 1981 tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag gtctccaaca 2041 aagccctccc agcccccatc gagaaaacca tctccaaagc caaagggcag ccccgagaac 2101 cacaggtgta caccctgccc ccatcccggg aggagatgac caagaaccag gtcagcctga 2161 cctgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag agcaatgggc 2221 agccggagaa caactacaag accacgcctc ccgtgctgga ctccgacggc tccttcttcc 2281 tctatagcaa gctcaccgtg gacaagagca ggtggcagca ggggaacgtc ttctcatgct 2341 ccgtgatgca tgaggctctg cacaaccact acacgcagaa gagcctctcc ctgtccccgg 2401 gtaaatgagt gcgacggccg gcaagccccc gctccccggg ctctcggggt cgcgcgagga 2461 aagcttggcc gccatggccc aacttgttta ttgcagctta taatggttac aaataaagca 2521 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 2581 ccaaactcat caatgtatct tatcatgtct ggatcgggaa ttaattcggc gcagcaccat 2641 ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc ggaaagaacc 2701 atctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa 2761 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc 2821 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc 2881 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct 2941 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga 3001 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctgtta acagcttggc 3061 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg 3121 ccttgcagca catccccctt tcgccagttg gcgtaatagc gaagaggccc gcaccgatcg 3181 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct 3241 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt 3301 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc 3361 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc 3421 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg 3481 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga 3541 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc 3601 caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg 3661 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt 3721 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc 3781 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt 3841 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag 3901 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt 3961 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga 4021 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc 4081 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt 4141 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct 4201 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt 4261 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt 4321 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac 4381 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac 4441 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct 4501 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg 4561 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg 4621 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca 4681 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa 4741 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt 4801 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc 4861 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg 4921 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt 4981 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt 5041 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc 5101 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 5161 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 5221 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 5281 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac 5341 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 5401 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 5461 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 5521 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 5581 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 5641 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 5701 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga g //