| 质粒名称: | pComb8 |
|---|---|
| 目录编号: | 63889 |
| 存储实验室: | Carlos Barbas |
|
载体骨架基本信息 |
|
| 载体名称: | pBluescript |
| 空载体大小: | 3300 bp |
| 完整质粒大小: | 3351bp |
| 载体类型: | Bacterial Expression ; phage display |
| 筛选标记: | geneVIII (C terminal on backbone) |
| 载体抗性: | Ampicillin, 100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | XL1 Blue |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 启动子: | lacZ |
| 克隆方法: | 限制性内切酶 |
|
文献引用方法 |
|
| 材料和方法部分: | pComb8 was a gift from Carlos Barbas (Addgene plasmid # 63889 ; http://n2t.net/addgene:63889 ; RRID:Addgene_63889) |
| 参考文献部分: | Assembly of combinatorial antibody libraries on phage surfaces: the gene III site. Barbas CF 3rd, Kang AS, Lerner RA, Benkovic SJ. Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):7978-82. PubMed 1896445 |
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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 pComb8 3351 bp ds-DNA circular SYN 14-OCT-2021
DEFINITION For phage display of antibody libraries by fusion to modified
geneVIII..
ACCESSION .
VERSION .
KEYWORDS pComb8
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3351)
AUTHORS Barbas CF 3rd, Kang AS, Lerner RA, Benkovic SJ
TITLE Assembly of combinatorial antibody libraries on phage surfaces: the
gene III site.
JOURNAL Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):7978-82.
PUBMED 1896445
REFERENCE 2 (bases 1 to 3351)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Oct 14, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..3351
/organism="synthetic DNA construct"
/mol_type="other DNA"
sig_peptide 40..105
/gene="pelB (fragment)"
/product="leader peptide for secretion"
/label=pelB signal sequence
primer_bind 119..135
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 120..136
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
sig_peptide 374..439
/gene="pelB (fragment)"
/product="leader peptide for secretion"
/label=pelB signal sequence
primer_bind complement(500..519)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(501..519)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(526..543)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(526..542)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(535..557)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin complement(684..1139)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(821..842)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 1033..1052
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 1166..1270
/gene="bla"
/label=AmpR promoter
CDS 1271..2131
/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(1489..1508)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2302..2890
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2791..2810
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3044..3061
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 3178..3199
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3214..3244
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3252..3268
/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 3257..3279
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3276..3292
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3276..3292
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 3313..3331
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
1 ggtggcggcc gcaaattcta tttcaaggag acagtcataa tgaaatacct attgcctacg
61 gcagccgctg gattgttatt actcgctgcc caaccagcca tggcccaggt gaaactgctc
121 gaggtcgacg gtatcgataa gcttgatatc gaattcctgc agcccggggg atccactagt
181 gctgagggtg acgatcccgc aaaagcggcc tttaactccc tgcaagcctc agcgaccgaa
241 tatatcggtt atgcgtgggc gatggttgtt gtcattgtcg gcgcaactat cggtatcaag
301 ctgtttaaga aattcacctc gaaagcaagc tgatagaatt ctaaactagc tagtcgccaa
361 ggagacagtc ataatgaaat acctattgcc tacggcagcc gctggattgt tattactcgc
421 tgcccaacca gccatggccg agctcgtcag ttctagagtt aagcggccgc aatcgagggg
481 gggcccggta cccaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt
541 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
601 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
661 gcgcagcctg aatggcgaat ggcaaattgt aagcgttaat attttgttaa aattcgcgtt
721 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
781 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
841 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
901 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
961 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
1021 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
1081 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
1141 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
1201 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
1261 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
1321 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
1381 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
1441 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
1501 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
1561 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
1621 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
1681 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
1741 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
1801 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
1861 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
1921 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
1981 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
2041 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
2101 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
2161 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
2221 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
2281 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
2341 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
2401 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
2461 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
2521 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
2581 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
2641 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
2701 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
2761 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
2821 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
2881 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
2941 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
3001 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
3061 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
3121 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
3181 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
3241 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
3301 cgccaagctc gaaattaacc ctcactaaag ggaacaaaag ctggccaccg c
//
