| 质粒名称: | pComb3XLambda |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 63892 |
| 存储实验室: | Carlos Barbas |
|
载体骨架基本信息 |
|
| 载体名称: | pComb3X |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 3300 bp |
| 完整质粒大小: | 4700 bp |
| 载体类型: | Bacterial Expression ; phage display |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | human antibody lambda light chain |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 320 bp |
| 启动子 | lacZ |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | SfiI(未破坏) |
| 3' 克隆位点: | SfiI (未破坏) |
| 5' 测序引物 | AAGACAGCTATCGCGATTGCAG |
| 3' 测序引物 | GCCCCCTTATTAGCGTTTGCCATC |
|
文献引用方法 |
|
| 材料和方法部分: | pComb3XLambda was a gift from Carlos Barbas (Addgene plasmid # 63892 ; http://n2t.net/addgene:63892 ; RRID:Addgene_63892) |
| 参考文献部分: | Methods for the generation of chicken monoclonal antibody fragments by phage display. Andris-Widhopf J, Rader C, Steinberger P, Fuller R, Barbas CF 3rd. J Immunol Methods. 2000 Aug 28;242(1-2):159-81. S0022175900002210 [pii] PubMed 10986398 |
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载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pComb3XLambda 4772 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Backbone pComb3X containing PCR template for human antibody lambda
light chain constant region.
ACCESSION .
VERSION .
KEYWORDS pComb3XLambda
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4772)
AUTHORS Andris-Widhopf J, Rader C, Steinberger P, Fuller R, Barbas CF 3rd
TITLE Methods for the generation of chicken monoclonal antibody fragments
by phage display.
JOURNAL J Immunol Methods. 2000 Aug 28;242(1-2):159-81.
PUBMED 10986398
REFERENCE 2 (bases 1 to 4772)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4772
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 109..130
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 145..175
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 183..199
/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)."
CDS 1743..1760
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 1767..1793
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
rep_origin complement(2387..2842)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2524..2545)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2736..2755
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2869..2973
/gene="bla"
/label=AmpR promoter
CDS 2974..3834
/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(3192..3211)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4005..4593
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4494..4513
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4747..4764
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
61 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
121 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
181 aattgtgagc ggataacaat tgaattcagg aggaatttaa aatgaaaaag acagctatcg
241 cgattgcagt ggcactggct ggtttcgcta ccgtggccca ggcggccgag ctcgtgctga
301 ctaacgccct cagtgtctgg ggccccaggg cagagggtca ccatctcctg cactgggagc
361 agctccaacc tcggggcagc ttatgatgta cactggtacc accaggttcc aggaacggcc
421 cccaaactcc tcctctatga taacatcaag cggccctcag gggtccctga ccgattctct
481 ggctccaagt ctggcacctc agcctccctg gccatcactg ggctccaggc tgaggacgag
541 gctgattatt actgcaacag agtaacagct ggcctcacac gttcggcgga gggaccaagc
601 tgaccgtcct aagtcagccc aaggctgccc cctcggtcac tctgttcccg ccctcctctg
661 aggagcttca agccaacaag gccacactgg tgtgtctcat aagtgacttc tacccgggag
721 ccgtgacagt ggcctggaag gcagatggca gccccgtcaa ggcgggagtg gagaccacca
781 caccctccaa acaaagcaac aacaagtacg cggccagcag ctatctgagc ctgacgcctg
841 agcagtggaa gtcccacaga agctacagct gccaggtcac gcatgaaggg agcaccgtgg
901 agaagacagt ggcccctaca gaatgttcat aattctagat aattaattag gaggaattta
961 aaatgaaata cctattgcct acggcagccg ctggattgtt attactcgct gcccaaccag
1021 ccatggccga ggtgcagctg ctcgagcagt ctggggctga ggtgaagaag cctgggtcct
1081 cggtgaaggt ctcctgcagg gcttctggag gcaccttcaa caattatgcc atcagctggg
1141 tgcgacaggc ccctggacaa gggcttgagt ggatgggagg gatcttccct ttccgtaata
1201 cagcaaagta cgcacaacac ttccagggca gagtcaccat taccgcggac gaatccacgg
1261 gcacagccta catggagctg agcagcctga gatctgagga cacggccata tattattgtg
1321 cgagagggga tacgattttt ggagtgacca tgggatacta cgctatggac gtctggggcc
1381 aagggaccac ggtcaccgtc tccgcagcct ccaccaaggg cccatcggtc ttccccctgg
1441 caccctcctc caagagcacc tctgggggca cagcggccct gggctgcctg gtcaaggact
1501 acttccccga accggtgacg gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca
1561 ccttcccggc tgtcctacag tcctcaggac tctactccct cagcagcgtg gtgaccgtgc
1621 cctccagcag cttgggcacc cagacctaca tctgcaacgt gaatcacaag cccagcaaca
1681 ccaaggtgga caagaaagtt gagcccaaat cttgtgacaa aactagtggc caggccggcc
1741 agcaccatca ccatcaccat ggcgcatacc cgtacgacgt tccggactac gcttcttagg
1801 agggtggtgg ctctgagggt ggcggttctg agggtggcgg ctctgaggga ggcggttccg
1861 gtggtggctc tggttccggt gattttgatt atgaaaagat ggcaaacgct aataaggggg
1921 ctatgaccga aaatgccgat gaaaacgtgc tacagtctga cgctaaaggc aaacttgatt
1981 ctgtcgctac tgattacggt gctgctatcg atggtttcat tggtgacgtt tccggccttg
2041 ctaatggtaa tggtgctact ggtgattttg ctggctctaa ttcccaaatg gctcaagtcg
2101 gtgacggtga taattcacct ttaatgaata atttccgtca atatttacct tccctccctc
2161 aatcggttga atgtcgccct tttgtctttg gcgctggtaa accatatgaa ttttctattg
2221 attgtgacaa aataaactta ttccgtggtg tctttgcgtt tcttttatat gttgccacct
2281 ttatgtatgt attttctacg tttgctaaca tactgcgtaa taaggagtct taagctagct
2341 aattaattta agcggccgca gatctgctct ctgaggagga tctgggaaat tgtaagcgtt
2401 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
2461 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
2521 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
2581 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
2641 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
2701 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
2761 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
2821 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
2881 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
2941 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
3001 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
3061 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
3121 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
3181 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
3241 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
3301 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
3361 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
3421 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
3481 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
3541 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
3601 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
3661 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
3721 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
3781 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
3841 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
3901 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
3961 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
4021 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
4081 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
4141 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
4201 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
4261 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
4321 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
4381 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
4441 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
4501 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
4561 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
4621 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
4681 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
4741 gagcgcagcg agtcagtgag cgaggaagcg ga
//
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