| 质粒名称: | pNIC-CH |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 26117 |
| 存储实验室: | Opher Gileadi |
|
载体骨架基本信息 |
|
| 载体名称: | pNIC-CH |
| 载体出品公司: | SGC Oxford |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 7218 bp |
| 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: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | T7-lacO |
| 标签/融合蛋白: | His（主干上的C终端） |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | T7 |
| 3' 测序引物 | T7-term |
|
文献引用方法 |
|
| 材料和方法部分: | pNIC-CH was a gift from Opher Gileadi (Addgene plasmid # 26117 ; http://n2t.net/addgene:26117 ; RRID:Addgene_26117) |
买家导航
pNICCH载体基本信息
pNICCH载体图谱质粒图谱和多克隆位点信息
pNICCH载体简介
pNICCH载体序列
LOCUS pNIC-CH 7218 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION SGC Empty backbone for bacterial expression.
ACCESSION .
VERSION .
KEYWORDS pNIC-CH
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7218)
TITLE pNIC-CH
REFERENCE 2 (bases 1 to 7218)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7218
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 150..169
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 253..272
/label=T7
/note="T7 promoter, forward primer"
promoter 253..271
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 272..296
/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 348..780
/gene="sacR"
/label=sacB promoter
/note="sacB promoter and control region"
CDS 781..2202
/codon_start=1
/gene="Bacillus subtilis sacB"
/product="secreted levansucrase that renders bacterial
growth sensitive to sucrose"
/label=SacB
/note="negative selection marker"
/translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
ITRHDMLQIPEQQKNEKYKVPEFDSSTIKNISSAKGLDVWDSWPLQNTDGTVANYHGYH
IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
VKDSILEQGQLTVNK"
CDS 2264..2281
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 2331..2348
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(2401..2419)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 2415..2462
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 2499..2954
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2586..2605)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2796..2817
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(3047..3862)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 3770..3789
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 3854..3873
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 3984..4572
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4473..4492
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4726..4743
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 4758..4900
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(4826..4845)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4986..5008
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(5002..5193)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(6002..7084)
/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 7046..7065
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(7085..7162)
/gene="lacI"
/label=lacI promoter
ORIGIN
1 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
61 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
121 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
181 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
241 atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac aattcccctc
301 tagaaataat tttgtttaac cttaagaagg agatatacta tgcaggtcgt tcactattat
361 ttagtgaaat gagatattat gatattttct gaattgtgat taaaaaggca actttatgcc
421 catgcaacag aaactataaa aaatacagag aatgaaaaga aacagataga ttttttagtt
481 ctttaggccc gtagtctgca aatcctttta tgattttcta tcaaacaaaa gaggaaaata
541 gaccagttgc aatccaaacg agagtctaat agaatgaggt cgaaaagtaa atcgcgcggg
601 tttgttactg ataaagcagg caagacctaa aatgtgtaaa gggcaaagtg tatactttgg
661 cgtcacccct tacatatttt aggtcttttt ttattgtgcg taactaactt gccatcttca
721 aacaggaggg ctggaagaag cagaccgcta acacagtaca taaaaaagga gacatgaacg
781 atgaacatca aaaagtttgc aaaacaagca acagtattaa cctttactac cgcactgctg
841 gcaggaggcg caactcaagc gtttgcgaaa gaaacgaacc aaaagccata taaggaaaca
901 tacggcattt cccatattac acgccatgat atgctgcaaa tccctgaaca gcaaaaaaat
961 gaaaaatata aagttcctga gttcgattcg tccacaatta aaaatatctc ttctgcaaaa
1021 ggcctggacg tttgggacag ctggccatta caaaacactg acggcactgt cgcaaactat
1081 cacggctacc acatcgtctt tgcattagcc ggagatccta aaaatgcgga tgacacatcg
1141 atttacatgt tctatcaaaa agtcggcgaa acttctattg acagctggaa aaacgctggc
1201 cgcgtcttta aagacagcga caaattcgat gcaaatgatt ctatcctaaa agaccaaaca
1261 caagaatggt caggttcagc cacatttaca tctgacggaa aaatccgttt attctacact
1321 gatttctccg gtaaacatta cggcaaacaa acactgacaa ctgcacaagt taacgtatca
1381 gcatcagaca gctctttgaa catcaacggt gtagaggatt ataaatcaat ctttgacggt
1441 gacggaaaaa cgtatcaaaa tgtacagcag ttcatcgatg aaggcaacta cagctcaggc
1501 gacaaccata cgctgagaga tcctcactac gtagaagata aaggccacaa atacttagta
1561 tttgaagcaa acactggaac tgaagatggc taccaaggcg aagaatcttt atttaacaaa
1621 gcatactatg gcaaaagcac atcattcttc cgtcaagaaa gtcaaaaact tctgcaaagc
1681 gataaaaaac gcacggctga gttagcaaac ggcgctctcg gtatgattga gctaaacgat
1741 gattacacac tgaaaaaagt gatgaaaccg ctgattgcat ctaacacagt aacagatgaa
1801 attgaacgcg cgaacgtctt taaaatgaac ggcaaatggt acctgttcac tgactcccgc
1861 ggatcaaaaa tgacgattga cggcattacg tctaacgata tttacatgct tggttatgtt
1921 tctaattctt taactggccc atacaagccg ctgaacaaaa ctggccttgt gttaaaaatg
1981 gatcttgatc ctaacgatgt aacctttact tactcacact tcgctgtacc tcaagcgaaa
2041 ggaaacaatg tcgtgattac aagctatatg acaaacagag gattctacgc agacaaacaa
2101 tcaacgtttg cgcctagctt cctgctgaac atcaaaggca agaaaacatc tgttgtcaaa
2161 gacagcatcc ttgaacaagg acaattaaca gttaacaaat aaaaacgcaa aagaaaatgc
2221 cgatatccta ttggcattga cgtcaggtgg cacacctgca gcgcaccatc atcaccacca
2281 ttgaggatcc gaattcgagc tccgtcgaca agcttgcggc cgcactcgag caccaccacc
2341 accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
2401 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
2461 tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt
2521 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
2581 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
2641 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
2701 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
2761 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
2821 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
2881 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
2941 aacgtttaca atttcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
3001 atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag aaaaactcat
3061 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
3121 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
3181 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
3241 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
3301 atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
3361 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
3421 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
3481 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
3541 ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
3601 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
3661 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
3721 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
3781 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta gagcaagacg
3841 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
3901 ttattgttca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
3961 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
4021 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
4081 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
4141 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
4201 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
4261 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
4321 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
4381 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
4441 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
4501 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
4561 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
4621 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
4681 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
4741 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
4801 caccgcatat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
4861 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
4921 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
4981 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
5041 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
5101 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
5161 catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct
5221 gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga
5281 tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg
5341 gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg
5401 tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg
5461 cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt
5521 tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga
5581 ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag
5641 cacgatcatg cgcacccgtg gggccgccat gccggcgata atggcctgct tctcgccgaa
5701 acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga ttccgaatac
5761 cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc cgaaaatgac
5821 ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag tcataagtgc
5881 ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa
5941 gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg
6001 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
6061 acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg
6121 agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt
6181 ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat
6241 aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca
6301 gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca
6361 tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg
6421 cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat
6481 gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga
6541 tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg
6601 agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat
6661 tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca
6721 gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc
6781 ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg
6841 ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca
6901 acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca
6961 tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc
7021 gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt
7081 tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg
7141 ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt
7201 aggaagcagc ccagtagt
//
