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pLLP-STII载体 pllp-stii质粒基本信息
pLLP-STII质粒图谱 pllp-stii载体图谱和pLLP-STII载体序列 pllp-stii质粒序列多克隆位点信息
pLLP-STII质粒 pllp-stii载体简介
pLLP-STII表达载体采用E.coli强启动lpp和STII分泌信号肽，同时在C端添加了6个His便于采用chelating sepharose亲和层析纯化目标蛋白。STII信号肽引导能力强且易于被E.coli信号肽酶切割。即使第一个氨基酸是半胱胺酸的干扰素也可以实现分泌表达。
pLLP-STII质粒序列 pllp-stii载体序列
LOCUS pllp-stii 7559 bp ds-DNA circular SYN 03-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pLLP-STII
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7559)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Tuesday, September 6, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..7559
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 247..277
/note="lac UV5 promoter"
/note="E. coli lac promoter with an ""up"" mutation"
protein_bind 285..301
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/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 309..325
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
RBS 361..383
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
misc_feature 424..459
/note="STll signal"
CDS 484..501
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
promoter 1866..1943
/gene="lacI"
/note="lacI promoter"
CDS 1944..3026
/codon_start=1
/gene="lacI"
/product="lac repressor"
/note="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"
protein_bind 3039..3060
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS 5125..5316
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 5418..5558
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(5744..6332)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6503..7363)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(7364..7468)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 ggataaccag aagcaataaa aaatcaaatc ggatttcact atataatctc actttatcta
61 agatgaatcc gatggaagca tcctgttttc tctcaatttt tttatctaaa acccagcgtt
121 cgatgcttct ttgagcgaac gatcaaaaat aagtgccttc ccatcaaaaa aatattctca
181 acataaaaaa ctttgtgtaa tacttgtaac gctacatgga gattaactca atctagctag
241 agaggcttta cactttatgc ttccggctcg tataatgtgt ggaattgtga gcggataaca
301 atttcacaca ggaaacagct atgaccatga ttacggattc actggaactc tagaaataat
361 tttgtttaac tttaagaagg agatatacat atgaaaaaga acatcgcatt cctcctggca
421 tctatgtttg ttttctctat cgctaccaac gcttacgctg gatccgcaga attcagcgct
481 agccaccatc atcatcatca ttaagcttag atccggctga gcaacgacgt gaacgcaatg
541 cgttccgacg ttcaggctgc taaagatgac gcagctcgtg ctaaccagcg tctggacaac
601 atggctacta aataccgcaa gtaatagtac ctgtgaagtg aaaaatggcg cacattgtgc
661 gacatttttt ttgtctgccg tttaccgcta ctgcgtcacg cgtaacatat tcccttgctc
721 tggttcacca ttctgcgctg actctactga aggcgcattg ctggctgcgg gagttgctcc
781 actgctcacc gaaaccggat accctgcccg acgatacaac gctttatcga ctaacttctg
841 atctacagcc ttattgtctt taaattgcgt aaagcctgct ggcagtgtgt atggcattgt
901 ctgaacgttc tgctgttctt ctgccgatag tggtcgatgt acttcaacat aacgcatccc
961 gttaggctcc acggaatatt tcaccggttc gttgatcact ttcaccggcg ttcccgtccg
1021 cacgctggag aacaaggctt taatatccgg tgcattcatg cgaatacacc ctgaactgac
1081 gcgcaaaccg acgctgtccg gcgcactggt accatgaatg aggtattcgc cattaccatg
1141 cgcgaggcgc agtgcgtaac gtcctagcgg gttatttggt ccggcaggaa cgactggcgg
1201 taatttaatg ccacgctcca gcgaacgctg acgaatgcct gccgtaggcg tccaggttgg
1261 gttagggatt ttctgcccaa cacgcgtttc catcaccggc gtttccagcc cctgcaatcc
1321 aatacctatt ggataaacct gcacaatatt ttctcccggc ggataataat aaaggcgcag
1381 ctctgcaagg ttggtcgacg gtcgaccggt ctcacttcaa cataacgcat cccgttaggc
1441 tccacggaat atttcaccgg ttcgttgatc actttcaccg gcgttcccgt ccgcacgctg
1501 gagaacaagg ctttaatatc cggtgcattc atgcgaatac accctgaact gacgcgcaaa
1561 ccgacgctgt ccggcgcact ggtaccatga atgaggtatt cgccattacc atgcgcgagg
1621 cgcagtgcgt aacgtcctag cgggttattt ggtccggcag gaacgactgg cggtaattta
1681 atgccacgct ccagcgaacg ctgacgaatg cctgccgtag gcgtccaggt tgggttaggg
1741 attttctgcc caacacgcgt ttccatcacc ggcgtttcca gcccctgcaa tccaatacct
1801 attggataaa cctgcacaat attttctccc ggcggataat aataaaggcg cagctctgca
1861 aggttgacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag
1921 agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg
1981 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
2041 aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg
2101 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
2161 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
2221 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
2281 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
2341 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
2401 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
2461 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
2521 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
2581 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
2641 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
2701 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
2761 acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt
2821 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
2881 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca
2941 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
3001 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
3061 taggcacccc aggctttaca ctttatgcta acgataatcc ctgacgcggt gcatcaggta
3121 ataacagttg tgaaggaata gttatcgtcg taccaggttt tggcaccggg gcgatagtgt
3181 tattggcttc aaggatcaac attgccgcag tatcaaaacg tcgggcaata gcctgaaggt
3241 ttttatcccc ttcttgcacc gtatacgttt gattttgccc aaccagtcgg cttccggttg
3301 gtggtagcgg ataatcaacc gcccaggcag cctggatggc gctaaaagcg ccgataagcg
3361 tgagtgtaag caaagacgcg cgtttcattg taaacctcct gtatttgccg gagactcacg
3421 ctgaaacgtc ggatggcgct tatgttcacc tgaaaccaaa acactcctgt gcaggtcagt
3481 gtaaacattg accatccggc aatgtgagcc aaccggatga aagctgtcct tttagtttag
3541 ctaagtgcag cggctttggc gcgaattgcg cgaatcatcg cttccagacc ttgtgaacga
3601 gatggggtga gatgttgggt gagcgccatt ttttcaaacc acggacgcac atcgaaattg
3661 acaatatcct gcggcgtcat ctgatcgtag agaataaaga cgaccgcaat aagccctttc
3721 acaatcgccg catcgctgtc gccctgtaat tcaataattc cctgggcatt ctggcgcatg
3781 acaatccaca cctgactctg acagccctga atgctatttt gtggacttct gtcttcgtcg
3841 cgtaattctg gcagacgctg ggggaccgat gcccttgaga gccttcaacc cagtcagctc
3901 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
3961 gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg
4021 ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc
4081 tcaagccttc gtcactggtc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc
4141 cggcatggcg gccgacgcgc tgggctacgt cttgctggcg ttcgcgacgc gaggctggat
4201 ggccttcccc attatgattc ttctcgcttc cggcggcatc gggatgcccg cgttgcaggc
4261 catgctgtcc aggcaggtag atgacgacca tcagggacag cttcaaggat cgctcgcggc
4321 tcttaccagc ctaacttcga tcactggacc gctgatcgtc acggcgattt atgccgcctc
4381 ggcgagcaca tggaacgggt tggcatggat tgtaggcgcc gccctatacc ttgtctgcct
4441 ccccgcgttg cgtcgcggtg catggagccg ggccacctcg acctgaatgg aagccggcgg
4501 cacctcgcta acggattcac cactccaaga attggagcca atcaattctt gcggagaact
4561 gtgaatgcgc aaaccaaccc ttggcagaac atatccatcg cgtccgccat ctccagcagc
4621 cgcacgcggc gcatctcggg cagcgttggg tcctggccac gggtgcgcat gatcgtgctc
4681 ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca
4741 ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca
4801 acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc
4861 tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct
4921 acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc
4981 atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca
5041 gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga
5101 aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc
5161 cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat
5221 gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc
5281 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
5341 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
5401 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
5461 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
5521 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
5581 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
5641 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
5701 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
5761 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
5821 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
5881 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
5941 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
6001 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
6061 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
6121 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
6181 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
6241 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
6301 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
6361 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
6421 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
6481 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
6541 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
6601 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
6661 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
6721 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
6781 cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct
6841 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
6901 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
6961 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
7021 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
7081 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac
7141 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
7201 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
7261 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
7321 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
7381 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
7441 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
7501 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggcccttt
//
