| 质粒名称: | pET22B_SA_PcrA |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 102999 |
| 存储实验室: | Martin Webb |
|
载体骨架基本信息 |
|
| 载体名称: | pET22B |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 30 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | SA_PcrA |
| 物种来源: | Staphylococcus aureus |
| 插入大小: | -- |
| 基因突变: | PcrA 199-2423 |
| GenBank编号 | -- |
| 启动子 | T7 |
| 标签/融合蛋白: | His tag（插入件上的N端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NdeI（销毁后未知） |
| 3' 克隆位点: | BamHI（销毁后未知） |
| 5' 测序引物 | T7 |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pET22B_SA_PcrA was a gift from Martin Webb (Addgene plasmid # 102999 ; http://n2t.net/addgene:102999 ; RRID:Addgene_102999) |
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 pET22B_SA_PcrA 7623 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION The plasmid is used to express Staphylococcus aureus PcrA, a DNA
helicase, with N-terminal His-tag and Gly-Gly linker.
ACCESSION .
VERSION .
KEYWORDS pET22B_SA_PcrA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7623)
TITLE Webb Lab plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7623)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7623
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 1..20
/label=T7
/note="T7 promoter, forward primer"
promoter 1..19
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 20..44
/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)."
RBS 59..81
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 92..109
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 2355..2372
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(2425..2443)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 2439..2486
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 2523..2978
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2610..2629)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2820..2841
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 3004..3108
/gene="bla"
/label=AmpR promoter
CDS 3109..3969
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(3327..3346)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4140..4728
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4629..4648
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4882..4899
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 4914..5053
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(4979..4998)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5139..5161
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(5155..5346)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(6155..7237)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 7199..7218
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(7238..7315)
/gene="lacI"
/label=lacI promoter
primer_bind 7521..7540
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
ORIGIN
1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
61 tgtttaactt taagaaggag atatacatat gcatcatcac catcaccacg gcggcatgaa
121 tgcactgctg aatcacatga ataccgaaca gagcgaagca gttaaaacca ccgaaggtcc
181 gctgctgatt atggcaggcg caggtagcgg taaaacccgt gttctgaccc atcgtattgc
241 atatctgctg gatgaaaaag atgtgagccc gtataatgtt ctggcaatta cctttaccaa
301 taaagcagcc cgtgaaatga aagaacgtgt tcagaaactg gttggtgatc aggccgaagt
361 tatttggatg agcacctttc atagcatgtg tgttcgtatt ctgcgtcgtg atgcagatcg
421 tattggtatt gaacgcaact ttaccattat tgatccgacc gatcagaaaa gcgttgttaa
481 agatgtgctg aaaaacgaga acatcgatag caaaaaattc gagcctcgca tgtttattgg
541 tgccattagc aatctgaaaa atgaactgaa aacaccggca gatgcacaga aagaagcaac
601 cgattatcat agccagatgg ttgccaccgt ttatagcggt tatcagcgtc agctgagccg
661 taatgaagca ctggattttg atgatctgat tatgaccacc atcaacctgt ttgaacgtgt
721 gccggaagtt ctggaatatt atcagaacaa attccagtat atccatgtgg atgagtatca
781 ggatacaaac aaagcacagt ataccctggt taaactgctg gcaagcaaat tcaaaaatct
841 gtgcgttgtt ggtgatagcg atcagagcat ttatggttgg cgtggtgcag atattcagaa
901 cattctgagc ttcgaaaaag attatccgga agccaatacc atctttctgg aacagaatta
961 tcgtagcacc aaaaccattc tgaatgcagc caatgaagtg atcaaaaata acagcgaacg
1021 taaaccgaaa ggtctgtgga ccgcaaatac caatggtgaa aaaatccact attatgaggc
1081 aatgaccgaa cgtgatgaag ccgaatttgt tattcgcgaa atcatgaaac atcagcgcaa
1141 cggcaaaaaa taccaggata tggcaattct gtatcgtacc aatgcacaga gccgtgtgct
1201 ggaagaaacc tttatgaaaa gcaatatgcc gtataccatg gtgggtggtc agaaattcta
1261 tgatcgcaaa gaaatcaaag acctgctgag ctatctgcgc attattgcaa atagcaacga
1321 tgatattagc ctgcagcgta ttatcaatgt tccgaaacgt ggtgttggtc cgagcagcgt
1381 tgaaaaagtt cagaattatg cactgcagaa caacatcagc atgtttgatg cactgggtga
1441 agcagatttc attggtctga gcaaaaaagt tacccaagaa tgcctgaact tctatgaact
1501 gattcagagc ctgatcaaag aacaagaatt cctggaaatc catgagatcg ttgatgaagt
1561 gctgcagaaa agtggttatc gtgaaatgct ggaacgtgaa aataccctgg aaagccgtag
1621 ccgtctggaa aatattgatg aatttatgag cgtgccgaaa gattatgaag aaaatacacc
1681 tctggaagaa cagtccctga ttaactttct gaccgatctg agcctggttg cagatatcga
1741 tgaagcggat accgaaaatg gtgttaccct gatgaccatg catagcgcaa aaggtctgga
1801 atttccgatt gtgtttatta tgggtatgga agaaagcctg tttccgcata ttcgtgcaat
1861 taaaagcgaa gatgatcacg agatgcaaga agaacgtcgt atttgttatg ttgcaattac
1921 ccgtgcagaa gaggttctgt atatcaccca tgcaaccagc cgtatgctgt ttggtcgtcc
1981 gcagagtaat atgccgagcc gttttctgaa agaaattccg gaaagcctgc tggaaaatca
2041 tagcagcggt aaacgtcaga ccattcagcc gaaagcaaaa ccgtttgcaa aacgtggttt
2101 tagccagcgt accaccagta ccaaaaaaca ggttctgagc agcgattgga atgtgggtga
2161 taaagtgatg cataaagcat ggggtgaagg tatggttagc aatgtgaatg aaaaaaacgg
2221 cagcatcgaa ctggacatca tctttaaaag ccagggtccg aaacggctgc tggcacagtt
2281 tgcaccgatt gaaaaaaaag aggactaagg atccgaattc gagctccgtc gacaagcttg
2341 cggccgcact cgagcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa
2401 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
2461 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg
2521 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
2581 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
2641 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
2701 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
2761 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
2821 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
2881 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
2941 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
3001 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
3061 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
3121 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
3181 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
3241 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
3301 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
3361 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
3421 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
3481 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
3541 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
3601 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
3661 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
3721 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
3781 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
3841 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
3901 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
3961 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
4021 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
4081 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
4141 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
4201 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
4261 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
4321 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
4381 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
4441 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
4501 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
4561 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
4621 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
4681 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
4741 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
4801 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
4861 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
4921 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
4981 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
5041 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
5101 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
5161 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
5221 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
5281 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
5341 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
5401 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
5461 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
5521 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
5581 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
5641 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
5701 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
5761 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
5821 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
5881 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
5941 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
6001 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
6061 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
6121 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
6181 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
6241 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
6301 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
6361 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
6421 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
6481 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
6541 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
6601 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
6661 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
6721 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
6781 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
6841 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
6901 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
6961 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
7021 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
7081 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
7141 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
7201 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
7261 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
7321 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
7381 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
7441 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
7501 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
7561 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
7621 aat
//
