质粒名称: | AbVec2.0-IGHG2 |
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出品公司: | Addgene |
目录编号: | 99576 |
存储实验室: | Vladislav Verkhusha |
载体骨架基本信息 |
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载体名称: | pUC19 |
载体出品公司: | Clontech |
空载体大小: | 5729bp |
完整质粒大小: | 5729bp |
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: | CMV |
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-IGHG2 was a gift from Hedda Wardemann (Addgene plasmid # 99576 ; http://n2t.net/addgene:99576 ; RRID:Addgene_99576)
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AbVec2.0-IGHG2载体质粒基本信息
AbVec2.0-IGHG2质粒图谱载体图谱和AbVec2.0-IGHG2载体序列质粒序列多克隆位点信息
AbVec2.0-IGHG2质粒载体简介
AbVec2.0-IGHG2质粒序列载体序列
LOCUS AbVec2.0-IGHG2 5729 bp DNA circular SYN 25-AUG-2024 DEFINITION Expression of secretory immunoglobulin heavy chains in mammalian cells, human IgG2 isotype. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5729) TITLE Immunoglobulin expression vectors JOURNAL Unpublished REFERENCE 2 (bases 1 to 5729) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.biofeng.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" FEATURES Location/Qualifiers source 1..5729 /mol_type="other DNA" /organism="synthetic DNA construct" protein_bind 98..119 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 134..164 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 172..188 /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 177..199 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 196..212 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 196..212 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" enhancer 241..620 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 621..824 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 775..795 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 821..845 /label=LNCX /note="Human CMV promoter, forward primer" promoter 1055..1073 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 1055..1072 /label=SP6 /note="SP6 promoter, forward primer" primer_bind 1086..1105 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 2256..2265 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" polyA_signal 2269..2403 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2306..2325) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2360..2379 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" promoter 2472..2829 /label=SV40 promoter /note="SV40 enhancer and early promoter" enhancer 2472..2663 /label=SV40 enhancer /note="enhancer for the SV40 early promoter (Herr, 1993)" primer_bind complement(2495..2515) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2680..2815 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(2849..2866) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2849..2865) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2858..2880) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3078..3533 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3165..3184) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3375..3396 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3550..3569) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3669..3691 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3729..3747) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3815..3919 /gene="bla" /label=AmpR promoter CDS 3920..4780 /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(4138..4157) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4951..5539 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5440..5459 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5693..5710 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atccaactgg cacgacaggt 61 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt 121 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg 181 gataacaatt tcacacagga aacagctatg acatgattac gaattaattc gagctcgccc 241 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 301 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 361 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 421 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 481 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 541 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 601 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 661 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 721 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 781 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc 841 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat 901 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg 961 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca 1021 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat 1081 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat 1141 cgattgaatt ccaccatggg atggtcatgt atcatccttt ttctagtagc aactgcaacc 1201 ggttgactaa ctagctggtc gaccaagggc ccaagcgtct tccccctggc gccctgctcc 1261 aggagcacct ccgagagcac agcggccctg ggctgcctgg tcaaggacta cttccccgag 1321 ccggtgacgg tgtcgtggaa ctcaggcgct ctgaccagcg gcgtgcacac cttcccagct 1381 gtcctacagt cctcaggact ctactccctc agcagcgtgg tgaccgtgcc ctccagcaac 1441 ttcggcaccc agacctacac ctgcaacgta gatcacaagc ccagcaacac caaggtggac 1501 aagacagttg agcgcaaatg ttgtgtcgag tgcccaccgt gcccagcacc acctgtggca 1561 ggaccgtcag tcttcctctt ccccccaaaa cccaaggaca ccctcatgat ctcccggacc 1621 cctgaggtca cgtgcgtggt ggtggacgtg agccacgaag accccgaggt ccagttcaac 1681 tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccacggga ggagcagttc 1741 aacagcacgt tccgtgtggt cagcgtcctc accgttgtgc accaggactg gctgaacggc 1801 aaggagtaca agtgcaaggt ctccaacaaa ggcctcccag cccccatcga gaaaaccatc 1861 tccaaaacca aagggcagcc ccgagaacca caggtgtaca ccctgccccc atcccgggag 1921 gagatgacca agaaccaggt cagcctgacc tgcctggtca aaggcttcta ccccagcgac 1981 atcgccgtgg agtgggagag caatgggcag ccggagaaca actacaagac cacacctccc 2041 atgctggact ccgacggctc cttcttcctc tacagcaagc tcaccgtgga caagagcagg 2101 tggcagcagg ggaacgtctt ctcatgctcc gtgatgcatg aggctctgca caaccactac 2161 acgcagaaga gcctctccct gtctccgggt aaatgagtgc cacggccggc aagcccccgc 2221 tccccaggct ctcggggtcg cgcgaggaaa gcttggccgc catggcccaa cttgtttatt 2281 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 2341 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg 2401 atcgggaatt aattcggcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg 2461 ttaggtacct tctgaggcgg aaagaaccat ctgtggaatg tgtgtcagtt agggtgtgga 2521 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 2581 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 2641 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc 2701 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 2761 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc 2821 ttttgcaaaa agctgttaac agcttggcac tggccgtcgt tttacaacgt cgtgactggg 2881 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagttggc 2941 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg 3001 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac 3061 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt 3121 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt 3181 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc 3241 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga 3301 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc 3361 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg 3421 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct 3481 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg 3541 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac 3601 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt 3661 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag 3721 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc 3781 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt 3841 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata 3901 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt 3961 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc 4021 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat 4081 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct 4141 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca 4201 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg 4261 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa 4321 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg 4381 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga 4441 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg 4501 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt 4561 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg 4621 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc 4681 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca 4741 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc 4801 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat 4861 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc 4921 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg 4981 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct 5041 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct 5101 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct 5161 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg 5221 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc 5281 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga 5341 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg 5401 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta 5461 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 5521 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 5581 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat 5641 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc 5701 agtgagcgag gaagcggaag agcgcccaa //
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AbVec2.0-IGHG2质粒载体应用举例
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