| 质粒名称: | pVDL9.3 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 168299 |
| 存储实验室: | Víctor de Lorenzo |
|
载体骨架基本信息 |
|
| 载体名称: | pHSG575 |
| 载体出品公司: | Tamotsu Hashimoto-Gotoh |
| 空载体大小: | 3600 bp |
| 完整质粒大小: | 7400 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 氯霉素25 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Segment of the hly operon, spanning the C-terminal part of hlyA, hlyB, and hlyD |
| 物种来源: | Synthetic |
| 插入大小: | 3739 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | LacI-Plac |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pVDL9.3 was a gift from Víctor de Lorenzo (Addgene plasmid # 168299 ; http://n2t.net/addgene:168299 ; RRID:Addgene_168299) |
| 参考文献部分: | An Escherichia coli hemolysin transport system-based vector for the export of polypeptides: export of Shiga-like toxin IIeB subunit by Salmonella typhimurium aroA. Tzschaschel BD, Guzman CA, Timmis KN, de Lorenzo V. Nat Biotechnol. 1996 Jun;14(6):765-9. doi: 10.1038/nbt0696-765. 10.1038/nbt0696-765 PubMed 9630987 |
买家导航
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 pVDL9.3 7313 bp ds-DNA circular SYN 17-JUN-2021
DEFINITION Secretion of polypeptides (nanobodies) in E. coli fused to
C-terminal secretion signal of HlyA.
ACCESSION .
VERSION .
KEYWORDS pVDL9.3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7313)
AUTHORS Tzschaschel BD, Guzman CA, Timmis KN, de Lorenzo V
TITLE An Escherichia coli hemolysin transport system-based vector for the
export of polypeptides: export of Shiga-like toxin IIeB subunit by
Salmonella typhimurium aroA.
JOURNAL Nat Biotechnol. 1996 Jun;14(6):765-9. doi: 10.1038/nbt0696-765.
PUBMED 9630987
REFERENCE 2 (bases 1 to 7313)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jun 17, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7313
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin 17..239
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
promoter 584..686
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 687..1346
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(753..772)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
primer_bind 1775..1797
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 1789..1806
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 1790..1806
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5692..5708)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5692..5708)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5705..5727)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5716..5732
/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)."
promoter complement(5740..5770)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5785..5806
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(6332..7282)
/codon_start=1
/gene="rep101"
/product="RepA protein needed for replication with the
pSC101 origin"
/label=Rep101
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
1 ctataccaat tgagatgggc tagtcaatga taattactag tccttttcct ttgagttgtg
61 ggtatctgta aattctgcta gacctttgct ggaaaacttg taaattctgc tagaccctct
121 gtaaattccg ctagaccttt gtgtgttttt tttgtttata ttcaagtggt tataatttat
181 agaataaaga aagaataaaa aaagataaaa agaatagatc ccagccctgt gtataactca
241 ctactttagt cagttccgca gtattacaaa aggatgtcgc aaacgctgtt tgctcctcta
301 caaaacagac cttaaaaccc taaaggctta agtagcaccc tcgcaagctc gggcaaatcg
361 ctgaatattc cttttgtctc cgaccatcag gcacctgagt cgctgtcttt ttcgtgacat
421 tcagttcgct gcgctcacgg ctctggcagt gaatgggggt aaatggcact acaggcgccg
481 aataaatacc tgtgacggaa gatcacttcg cagaataaat aaatcctggt gtccctgttg
541 ataccgggaa gccctgggcc aacttttggc gaaaatgaga cgttgatcgg cacgtaagag
601 gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga
661 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
721 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
781 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
841 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
901 aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt
961 acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg
1021 atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg
1081 cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga
1141 gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca
1201 ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc
1261 atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact
1321 gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc
1381 ctggttgcta cgcctgaata agtgataata agcggatgaa tggcagaaat tcgaaagcaa
1441 attcgacccg gtcgtcggtt cagggcaggg tcgttaaata gccgcttatg tctattgctg
1501 gtttaccggt ttattgacta ccggaagcag tgtgaccgtg tgcttctcaa atgcctgagg
1561 ccagtttgct caggctctcc ccgtggaggt aataattgac gatatgatca tttattctgc
1621 ctcccagagc ctgataaaaa cggttagcgc cattcgccat tcaggctgcg caactgttgg
1681 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
1741 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
1801 gccagtgcca agctgggcgg ccgcgcgaat tccagtggat agtcagtact ggtttgccac
1861 aaaacagtgc agtcacacat gacaggagaa gatatgagcc ggataccgcg gctctgagac
1921 ttaacgctca tgtaaacttt ctgttacaga ctcttccaga ggactaagaa gatagctgat
1981 tacgcttcgc attccagtct ttatttctgc agtgacagcc atacccgagc ttaatggaat
2041 gtgcttattc ccggttgaca aatcattctc ttcaacagaa acaatgacat taaaaacgag
2101 tcccagtttc tggtcttcta ttgcatctaa atttatattt ttcaccttac ccaccagata
2161 accatatcgg gtgtaaggaa aggcctccac tttaatgatg gcattctgcc cgacgttaat
2221 aaaaccaata tctttatttt gtaccagagc agtaacctcc agcgtgtcat cttccggaac
2281 gatgaccatc agtgtttccg ctgttgtaac aaccccacct tcagtatgaa ccttcagttg
2341 ctgaactttt cccgaaacag gggccctgat tactgaagcc tgttgacgct cttcattttt
2401 ctctaactcc agagttaata actcaatgct gtctgttgtt tgtcttagct tgtctaaaat
2461 ttcattttta aaaagctgcg tgacaagctg atattcttct tttgcagaca atatctcact
2521 ctcaatttgc tccagttgcg atttataaac ccgtaattca tttgctgcct cgacatattt
2581 attctcctgc tcaagtacag catgttttgc aattgcctgt ttatgcaata aactcctgaa
2641 atcatccaga cggctttttt caactctcga taaattttca taacggttta tacgggcaag
2701 tattgttaat cgctctgctc ttttcttatc cagattcagt tctttttgat acttctgatt
2761 ttgccatgtg gaaaactgtt cttttatcaa agaagttaaa cgcagtactt cctcttcaga
2821 tacattctga aaataaggct catcaggaag cttcagttca ggtagtttat ttaattcaat
2881 tgacctgctc agaatttgat accgagtttg ttccagcctg gtctgtaaca gtgatgactg
2941 tgtttttaac gtatcagctt cagctcccag tgctgtaagc tttaataaca catccccttt
3001 ccggactgac tctccttctt ttacgataat ttctttaact attgagtttt caataggttt
3061 aatttctttg ctacgcccac ttagtgttaa tttcccattt gcagtggcaa caatttccac
3121 ctgacctaaa acagataaaa tgacagcaat aaccagaaac cccataataa aataagcaac
3181 cagacgcggc cgtctggata ccggcgtttc aattaattcc agatgagcgg gtaagaattc
3241 attttcgtcc ttttcacgta ccggagtatc taattgcttc cggattttcc atgtttcact
3301 ccagacaagt ttatagcaca acaggaactc gctgaacccc attaaccatg ttttcatatt
3361 cttctgttct ttctgttagt ctgactgtaa ctgatataag taactgtata aactttccgg
3421 ttcagaaagc agttccttat gtttaccctg ttcaacaatt ttcccttttt ccatgacaat
3481 aatgcggtct gcatttttta ctgtagacag acgatgagca atgattataa ccgttctgcc
3541 cttacatatt ttgtgcatat tgcgcatgat gatatgctcc gactcataat ccagagcact
3601 ggttgcttca tcaaaaataa gtattttagg gttgttcacc agcgcccttg caattgcgat
3661 gcgttgacgt tgacctccgg ataatcctgc cccctgttcc ccgacaatgg tgttataccc
3721 ctcacgcaat tcagaaataa aatcatgagc gcctgctaat ttcgctgcat aaataacttt
3781 ttcgacggac ataccaggat tagccagtga gatattatca ataatactgc gattaagcag
3841 cacattgtcc tgcaacacaa cccccacctg acgacgtaac cagttaggat cggccaacgc
3901 aagatcatgt ccatcaatta agacctggcc attttcagga atataaaaac gttgaattaa
3961 tttagttaat gtgctttttc ctgaaccaga acgtccgaca ataccaataa cctccccctg
4021 cttaatactg agattgatat tatctaaaat aaccggagag tcaggcttat agcgaaaccg
4081 gatattacga aaagtgatat caccattaat ttccggtaat gccagtttcc catgataact
4141 ttcagttgga gagttaagca catcaccaag gcgggtaact gatataccaa cctgctggaa
4201 atcctgccag atttgtgcaa ggcgaataac cggtgcaaca atctgaccag caagcatatt
4261 aaaagcaatt aactgaccaa tacttaaatc cccggaaata accaggtgtg ctcccaacca
4321 caggttgatg atcataacag tcttttgtat taactgtatt ccttgttgac caatggttgc
4381 taatactgtc actttgaagc ctgcagcaac atatcctgcc aattgtttgt cccatatgtt
4441 cgtcatctga ggtgagactg ccatagcttt tatagtgtta atcgccgtga ctgattccac
4501 caggaaagat tgattatccg cattccgtga aaacttatca tcaaggcgac gtcgcaaaat
4561 ggggctaata aaaacagacc atgcagcata acaaggcagc gaaaataaga tcaccagagt
4621 aagctttgga ctgtaatacc acattaccgc aaaaaatatg aatgaaaata ataagtccag
4681 aacagatgtt aatgcctgtc ctgtcagaaa attacggatc tggtctaatt ctcttaccct
4741 ggcaacagta tcaccaacac gacgactctc aaaataagag atcggtagcg ccagtaaatg
4801 ccggaagagt ttggcaccca actcaacatc aatccgactt gtactatgtg caaaaatgta
4861 agttcttaaa ccgctgagta taatctcaaa caccaccaca acagataatg cgacagtaat
4921 aacattaaga gttgaaaatc ccctgtgcac taatactttg tccataacca cctgaaaaaa
4981 aaggggggtt attaatgcaa ataattgtaa aaaaacagac acaacaaggg tttcaataaa
5041 tattctcctg tattttataa tggcaggaat aaaccaggta aagtcaaatt tcgccagttt
5101 cccggcaaca gaagaacggg aagcgataag aataatatgc ccctgatata acgcctcaaa
5161 ctcagactgt tcgagaacac ggggatttcg ctgctccaga tcaaaaataa gatatctgtt
5221 tgcttcttta ctgactttag tcagaataaa atgacgtcca tcctctctcc agactaatgc
5281 gggcagagaa ataaagttta atcggtcaat tgtttttttt acctgtttta cctttagttc
5341 taaagatttc gcagcaagca accatgacgt taatcccaga ccagtcccgt ctgtgtcaaa
5401 tctatgttta atttcttccg ggttaacaga gacgttatgg tattgggcta aaatctccag
5461 ggcgtataac ccataatcaa ttttatgaca agaatccatt atgactccaa aaaaatagca
5521 atcttatgtg gcacagccca gtaagattgc tatcatttaa attaatatat tatgctgatg
5581 ctgtcaaagt tattgagttc cgtccatatg aaaaatcact ggcattaccg gacaactgca
5641 ataaagaagc ggcagatctt cctagcacag cggccgcaat tcgtaatcat ggtcatagct
5701 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
5761 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
5821 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
5881 cgcggggaga ggcggtttgc gtattgggcg ctatttcttc cagaattgcc atgatttttt
5941 ccccacggga ggcgtcactg gctcccgtgt tgtcggcagc tttgattcga taagcagcat
6001 cgcctgtttc aggctgtcta tgtgtgactg ttgagctgta acaagttgtc tcaggtgttc
6061 aatttcatgt tctagttgct ttgttttact ggtttcacct gttctattag gtgttacatg
6121 ctgttcatct gttacattgt cgatctgttc atggtgaaca gctttaaatg caccaaaaac
6181 tcgtaaaagc tctgatgtat ctatcttttt tacaccgttt tcatctgtgc atatggacag
6241 ttttcccttt gatatgtaac ggtgaacagt tgttctactt ttgtttgtta gtcttgatgc
6301 ttcactgata gatacaagag ccataagaac ctcagatcct tccgtattta gccagtatgt
6361 tctctagtgt ggttcgttgt ttttgcgtga gccatgagaa cgaaccattg agatcatact
6421 tactttgcat gtcactcaaa aattttgcct caaaactggt gagctgaatt tttgcagtta
6481 aagcatcgtg tagtgttttt cttagtccgt tatgtaggta ggaatctgat gtaatggttg
6541 ttggtatttt gtcaccattc atttttatct ggttgttctc aagttcggtt acgagatcca
6601 tttgtctatc tagttcaact tggaaaatca acgtatcagt cgggcggcct cgcttatcaa
6661 ccaccaattt catattgctg taagtgttta aatctttact tattggtttc aaaacccatt
6721 ggttaagcct tttaaactca tggtagttat tttcaagcat taacatgaac ttaaattcat
6781 caaggctaat ctctatattt gccttgtgag ttttcttttg tgttagttct tttaataacc
6841 actcataaat cctcatagag tatttgtttt caaaagactt aacatgttcc agattatatt
6901 ttatgaattt ttttaactgg aaaagataag gcaatatctc ttcactaaaa actaattcta
6961 atttttcgct tgagaacttg gcatagtttg tccactggaa aatctcaaag cctttaacca
7021 aaggattcct gatttccaca gttctcgtca tcagctctct ggttgcttta gctaatacac
7081 cataagcatt ttccctactg atgttcatca tctgagcgta ttggttataa gtgaacgata
7141 ccgtccgttc tttccttgta gggttttcaa tcgtggggtt gagtagtgcc acacagcata
7201 aaattagctt ggtttcatgc tccgttaagt catagcgact aatcgctagt tcatttgctt
7261 tgaaaacaac taattcagac atacatctca attggtctag gtgattttaa tca
//
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