pET28a-SUMO(pET28aSUMO)

pET28a-SUMO(pET28aSUMO)

质粒名称: pET28a-SUMO
其他名称:
质粒大小（bp）:5633bp
抗性:卡纳霉素
质粒类型:大肠杆菌表达载体
是否病毒:
启动子:T7
稳转或瞬转:
组成型或诱导性:
表达水平:
蛋白标签:N-His, N-Thrombin, N-T7,N-SUMO, C-His
5′ 测序引物及序列:T7：TAATACGACTCACTATAGGG

3′ 测序引物及序列：T7-Ter：TGCTAGTTATTGCTCAGCGG
GenBank号:
其他属性:
质粒图谱:

质粒序列:LOCUS       Exported                5633 bp ds-DNA     circular SYN 03-8-2015
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 5
SOURCE      synthetic DNA construct
ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5633)
AUTHORS   .
TITLE     Direct Submission
JOURNAL   Exported 2015-8-3   from MLCC
http://www.miaolingbio.com
FEATURES             Location/Qualifiers
source          1..5633
/organism=”synthetic DNA construct”
/mol_type=”other DNA”
source          5042..5064
/organism=”Enterobacteria phage T7″
/mol_type=”genomic DNA”
/db_xref=”taxon:10760″
rep_origin      12..467
/direction=RIGHT
/note=”f1 ori”
/note=”f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis”
CDS             complement(560..1375)
/codon_start=1
/product=”aminoglycoside phosphotransferase”
/note=”KanR”
/note=”confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes”
/translation=”MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF”
rep_origin      1497..2085
/direction=RIGHT
/note=”ori”
/note=”high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication”
misc_feature    2271..2413
/note=”bom”
/note=”basis of mobility region from pBR322″
CDS             complement(2515..2706)
/codon_start=1
/gene=”rop”
/product=”Rop protein, which maintains plasmids at low copy
number”
/note=”rop”
/translation=”MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL”
CDS             complement(3515..4597)
/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”
promoter        complement(4598..4675)
/gene=”lacI”
/note=”lacI promoter”
promoter        4984..5002
/note=”T7 promoter”
/note=”promoter for bacteriophage T7 RNA polymerase”
protein_bind    5003..5027
/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).”
RBS             5042..5064
/note=”efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)”
CDS             5083..5100
/codon_start=1
/product=”6xHis affinity tag”
/note=”6xHis”
/translation=”HHHHHH”
CDS             5110..5127
/codon_start=1
/product=”thrombin recognition and cleavage site”
/note=”thrombin site”
/translation=”LVPRGS”
CDS             5140..5430
/codon_start=1
/gene=”S. cerevisiae SMT3 (truncated)”
/product=”cleavable ubiquitin-like protein tag”
/note=”SUMO”
/translation=”MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIG”
CDS             5477..5494
/codon_start=1
/product=”6xHis affinity tag”
/note=”6xHis”
/translation=”HHHHHH”
terminator      5561..5608
/note=”T7 terminator”
/note=”transcription terminator for bacteriophage T7 RNA
polymerase”
ORIGIN
1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
541 tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat
601 tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa
661 actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc
721 gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga
781 aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc
841 agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac
901 cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac
961 aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat
1021 tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag
1081 tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca
1141 taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac
1201 ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg
1261 tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca
1321 tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac
1381 cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa
1441 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
1501 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
1561 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
1621 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
1681 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
1741 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
1801 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
1861 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
1921 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
1981 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
2041 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
2101 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
2161 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
2221 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
2281 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta
2341 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
2401 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
2461 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
2521 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
2581 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
2641 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
2701 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
2761 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
2821 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
2881 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
2941 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
3001 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
3061 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
3121 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
3181 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
3241 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
3301 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
3361 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
3421 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
3481 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
3541 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
3601 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
3661 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
3721 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
3781 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
3841 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
3901 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
3961 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
4021 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
4081 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
4141 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
4201 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
4261 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
4321 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
4381 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
4441 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
4501 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
4561 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
4621 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
4681 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
4741 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
4801 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
4861 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
4921 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
4981 aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa
5041 ttttgtttaa ctttaagaag gagatatacc atgggcagca gccatcatca tcatcatcac
5101 agcagcggcc tggtgccgcg cggcagccat atggctagca tgtcggactc agaagtcaat
5161 caagaagcta agccagaggt caagccagaa gtcaagcctg agactcacat caatttaaag
5221 gtgtccgatg gatcttcaga gatcttcttc aagatcaaaa agaccactcc tttaagaagg
5281 ctgatggaag cgttcgctaa aagacagggt aaggaaatgg actccttaag attcttgtac
5341 gacggtatta gaattcaagc tgatcagacc cctgaagatt tggacatgga ggataacgat
5401 attattgagg ctcacagaga acagattggt ggatccgaat tcgagctccg tcgacaagct
5461 tgcggccgca ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg
5521 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
5581 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gat