| 质粒名称: | AbVec1.1-IGKC |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 80796 |
| 存储实验室: | Hedda Wardemann |
|
载体骨架基本信息 |
|
| 载体名称: | pUC19 |
| 空载体大小: | 5020 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 |
|
| 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 |
|
| 材料和方法部分: | AbVec1.1-IGKC was a gift from Hedda Wardemann (Addgene plasmid # 80796 ; http://n2t.net/addgene:80796 ; RRID:Addgene_80796) |
| 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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AbVec1.1-IGKC载体 AbVec1.1IGKC质粒基本信息
AbVec1.1-IGKC质粒图谱 AbVec1.1IGKC载体图谱和AbVec1.1-IGKC载体序列 AbVec1.1IGKC质粒序列多克隆位点信息
AbVec1.1-IGKC质粒 AbVec1.1IGKC载体简介
AbVec1.1-IGKC质粒序列 AbVec1.1IGKC载体序列
LOCUS AbVec1.1-IGKC 5020 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expression of immunoglobulin light chains in mammalian cells, human
kappa constant region.
ACCESSION .
VERSION .
KEYWORDS AbVec1.1-IGKC
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5020)
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 5020)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5020
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 92..113
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 128..158
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 166..182
/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 171..193
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 190..206
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 190..206
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 815..839
/label=LNCX
/note="Human CMV promoter, forward primer"
promoter 1049..1067
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 1049..1066
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind 1080..1099
/label=pMT2-F
/note="Synthetic intron, forward primer"
regulatory 1541..1550
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
polyA_signal 1554..1688
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1591..1610)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1645..1664
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
promoter 1757..2114
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(1780..1800)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 1965..2100
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2027..2046
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(2134..2151)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(2134..2150)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2143..2165)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 2363..2818
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2450..2469)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2660..2681
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2835..2854)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2954..2976
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(3014..3032)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3100..3204
/gene="bla"
/label=AmpR promoter
CDS 3205..4065
/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(3423..3442)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4236..4824
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4725..4744
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4978..4995
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 aaccgcctct ccccgcgcgt tggccgattc attaatccaa ctggcacgac aggtttcccg
61 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
121 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
181 aatttcacac aggaaacagc tatgacatga ttacgaatta attcgagctc gcccgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg
841 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
901 tccgcggccg ggaacggtgc attggaacgc ggattccccg tgccaagagt gacgtaagta
961 ccgcctatag agtctatagg cccaccccct tggcttcgtt agaacgcggc tacaattaat
1021 acataacctt atgtatcata cacatacgat ttaggtgaca ctatagaata acatccactt
1081 tgcctttctc tccacaggtg tccactccca ggtccaactg cacctcggtt ctatcgattg
1141 aattccacca tgggatggtc atgtatcatc ctttttctag tagcaactgc aaccggttga
1201 ctaactagcc gtacggtggc tgcaccatct gtcttcatct tcccgccatc tgatgagcag
1261 ttgaaatctg gaactgcctc tgttgtgtgc ctgctgaata acttctatcc cagagaggcc
1321 aaagtacagt ggaaggtgga taacgccctc caatcgggta actcccagga gagtgtcaca
1381 gagcaggaca gcaaggacag cacctacagc ctcagcagca ccctgacgct gagcaaagca
1441 gactacgaga aacacaaagt ctacgcctgc gaagtcaccc atcagggcct gagctcgccc
1501 gtcacaaaga gcttcaacag gggagagtgt tagaagcttg gccgccatgg cccaacttgt
1561 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
1621 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
1681 tctggatcgg gaattaattc ggcgcagcac catggcctga aataacctct gaaagaggaa
1741 cttggttagg taccttctga ggcggaaaga accatctgtg gaatgtgtgt cagttagggt
1801 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
1861 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
1921 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
1981 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
2041 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
2101 taggcttttg caaaaagctg ttaacagctt ggcactggcc gtcgttttac aacgtcgtga
2161 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
2221 ttggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
2281 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
2341 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
2401 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
2461 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
2521 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
2581 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
2641 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
2701 tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
2761 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
2821 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
2881 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
2941 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
3001 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
3061 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
3121 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
3181 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
3241 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
3301 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
3361 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
3421 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
3481 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
3541 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
3601 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
3661 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
3721 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
3781 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
3841 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
3901 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
3961 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
4021 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
4081 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
4141 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
4201 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
4261 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
4321 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
4381 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
4441 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
4501 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
4561 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
4621 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
4681 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
4741 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
4801 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
4861 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
4921 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
4981 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
//
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