| 質粒名稱: | AbVec2.0-IGHG1 |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 80795 |
| 存儲實驗室: | Hedda Wardemann |
|
載體骨架基本信息 |
|
| 載體名稱: | pUC19 |
| 空載體大小: | 5741 bp |
| 載體類型: | 哺乳細胞表達 |
| 載體抗性: | 氨芐霉素 |
| 生長溫度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷貝數: | 高拷貝 |
|
插入基因信息 |
|
| 啟動子: | hCMV IE1 gene promoter |
| 克隆方法: | 限制性內切酶 |
| 5’測序引物 | GCTTCGTTAGAACGCGGCTAC |
|
文獻引用方法 |
|
| 材料和方法部分: | AbVec2.0-IGHG1 was a gift from Hedda Wardemann (Addgene plasmid # 80795 ; //n2t.net/addgene:80795 ; RRID:Addgene_80795) |
| 參考文獻部分: | 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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AbVec2.0-IGHG1載體 AbVec2.0IGHG1質粒基本信息
AbVec2.0-IGHG1質粒圖譜 AbVec2.0IGHG1載體圖譜和AbVec2.0-IGHG1載體序列 AbVec2.0IGHG1質粒序列多克隆位點信息
AbVec2.0-IGHG1質粒 AbVec2.0IGHG1載體簡介
AbVec2.0-IGHG1質粒序列 AbVec2.0IGHG1載體序列
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
//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
//