pET-47b(+)

pET-47b(+)

质粒名称: pET-47b(+)
其他名称:
质粒大小（bp）: 5203
抗性: Kanamycin (卡那霉素)
质粒类型: 大肠杆菌表达载体
是否病毒: Nonviral
启动子:
稳转或瞬转: Transient
组成型或诱导性: Constitutive
表达水平: High
蛋白标签: N-His, C-S, C-Thrombin
5′ 测序引物序列:T7: 5′-TAATACGACTCACTATAGGG-3′

3′ 测序引物序列: T7-ter:TGCTAGTTATTGCTCAGCGG
GenBank号:
其他属性: Contains HRV 3C Protease cleavage site for fusion tag removal a low temperatures; Cterm thrombin cleavage site
质粒图谱:

质粒序列：

LOCUS       Exported                5203 bp ds-DNA     circular SYN 06-SEP-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pET-47b
SOURCE      synthetic DNA construct
ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5203)
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..5203
/organism=”synthetic DNA construct”
/mol_type=”other DNA”
terminator      26..73
/note=”T7 terminator”
/note=”transcription terminator for bacteriophage T7 RNA
polymerase”
CDS             complement(168..212)
/codon_start=1
/product=”affinity and epitope tag derived from pancreatic
ribonuclease A”
/note=”S-Tag”
/translation=”KETAAAKFERQHMDS”
CDS             complement(213..230)
/codon_start=1
/product=”thrombin recognition and cleavage site”
/note=”thrombin site”
/translation=”LVPRGS”
CDS             complement(309..332)
/codon_start=1
/product=”recognition and cleavage site for human
rhinovirus 3C and PreScission proteases”
/note=”HRV 3C site”
/translation=”LEVLFQGP”
CDS             complement(342..359)
/codon_start=1
/product=”6xHis affinity tag”
/note=”6xHis”
/translation=”HHHHHH”
protein_bind    410..434
/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).”
promoter        complement(435..453)
/note=”T7 promoter”
/note=”promoter for bacteriophage T7 RNA polymerase”
promoter        766..843
/gene=”lacI”
/note=”lacI promoter”
CDS             844..1926
/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”
CDS             2498..2689
/codon_start=1
/gene=”rop”
/product=”Rop protein”
/note=”rop”
/translation=”MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL”
rep_origin      complement(3119..3707)
/direction=LEFT
/note=”ori”
/note=”high-copy-number colE1/pMB1/pBR322/pUC origin of
replication”
CDS             3829..4644
/codon_start=1
/gene=”aph(3′)-Ia”
/product=”aminoglycoside phosphotransferase”
/note=”KanR”
/note=”confers resistance to kanamycin”
/translation=”MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF”
rep_origin      complement(4737..5192)
/direction=LEFT
/note=”f1 ori”
/note=”f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis”
ORIGIN
1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
121 tgtttagcag cctaggttaa ttaagcctcg agagcagcag aagtagagct gtccatgtgc
181 tggcgttcga atttagcagc agcggtttct ttactaccgc gtggcaccag agcgagctct
241 gcggccgcaa gcttgtcgac ggacgtcggg cgcgccaagg cctgtacaga attcggatcc
301 tggtacccgg gtccctgaaa gaggacttca agagccgcgg agtgatggtg gtggtgatgt
361 gccatatgta tatctccttc ttaaagttaa acaaaattat ttctagaggg gaattgttat
421 ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tcgatctcga
481 tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg
541 cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg
601 cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct
661 ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct
721 gcttcctaat gcaggagtcg cataagggag agcgtcgaga tcccggacac catcgaatgg
781 cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
841 aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc
901 gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa
961 gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa
1021 cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt
1081 gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta
1141 gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc
1201 agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc
1261 tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt
1321 attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac
1381 cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct
1441 ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac
1501 tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc
1561 actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag
1621 tccgggctgc gcgttggtgc ggacatctcg gtagtgggat acgacgatac cgaagacagc
1681 tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc
1741 agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg
1801 cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc
1861 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
1921 cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcaccgg gatctcgacc
1981 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactag
2041 catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta
2101 gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc
2161 gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg
2221 gaagtcagcg ccctgcacca ttatgttccg gatctgcatc gcaggatgct gctggctacc
2281 ctgtggaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag tgatttttct
2341 ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca gtaaccgggc
2401 atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg gtatcattac
2461 ccccatgaac agaaatcccc cttacacgga ggcatcagtg accaaacagg aaaaaaccgc
2521 ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga
2581 gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct
2641 ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
2701 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
2761 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
2821 cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
2881 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
2941 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
3001 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
3061 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
3121 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
3181 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
3241 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
3301 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
3361 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
3421 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
3481 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
3541 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
3601 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
3661 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
3721 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
3781 tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt
3841 caacgggaaa cgtcttgctc taggccgcga ttaaattcca acatggatgc tgatttatat
3901 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat
3961 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
4021 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
4081 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggcaaa
4141 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
4201 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagtgat
4261 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
4321 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
4381 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
4441 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
4501 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
4561 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
4621 ttgatgctcg atgagttttt ctaagaatta attcatgagc ggatacatat ttgaatgtat
4681 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgaaat
4741 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
4801 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
4861 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
4921 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
4981 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
5041 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
5101 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
5161 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg cca