质粒序列
LOCUS Exported 5598 bp ds-DNA circular SYN 20-七月-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pSUMO
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5598)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2016年9月18日 from SnapGene Viewer 2.8.1
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..5598
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 26..73
/note="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(140..157)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(203..493)
/codon_start=1
/gene="S. cerevisiae SMT3 (truncated)"
/product="cleavable ubiquitin-like protein tag"
/note="SUMO"
/translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
RLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
CDS complement(497..514)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
RBS 527..549
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 564..588
/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(589..607)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 916..993
/gene="lacI"
/note="lacI promoter"
CDS 994..2076
/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 2885..3076
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 3178..3320
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(3506..4094)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 4225..5040
/codon_start=1
/gene="aph(3')-Ia"
/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 complement(5132..5587)
/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 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
181 cgacggagct cgaattcgga tcacctccaa tctgttctct gtgagcctca ataatatcgt
241 tatcctccat gtccaaatct tcaggggtct gatcagcttg aattctaata ccgtcgtaca
301 agaatcttaa ggagtccatt tccttaccct gtcttttagc gaacgcttcc atcagccttc
361 ttaaaggagt ggtctttttg atcttgaaga agatctctga agatccatcg gacaccttta
421 aattgatgtg agtctcaggc ttgacttctg gcttgacctc tggcttagct tcttgattga
481 cttctgagtc cgacccgtga tgatgatggt gatgacccat ggtatatctc cttcttaaag
541 ttaaacaaaa ttatttctag aggggaattg ttatccgctc acaattcccc tatagtgagt
601 cgtattaatt tcgcgggatc gagatctcga tcctctacgc cggacgcatc gtggccggca
661 tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag
721 atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc
781 ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg
841 tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag
901 agcgtcgaga tcccggacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag
961 cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc
1021 gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac
1081 gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc
1141 aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc
1201 agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa
1261 ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg
1321 gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat
1381 gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat
1441 gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg
1501 ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta
1561 agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa
1621 attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc
1681 atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg
1741 gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg
1801 gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc
1861 aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag
1921 ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc
1981 ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
2041 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta
2101 gctcactcat taggcaccgg gatctcgacc gatgcccttg agagccttca acccagtcag
2161 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
2221 catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt
2281 tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct
2341 cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat
2401 cgccggcatg gcggccccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct
2461 aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg
2521 aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt
2581 ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat
2641 ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg
2701 ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt
2761 accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat
2821 cctctctcgt ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc
2881 atcagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac
2941 attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga
3001 atcgcttcac gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga
3061 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
3121 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
3181 agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca
3241 gagcagattg tactgagagt gcaccatata tgcggtgtga aataccgcac agatgcgtaa
3301 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
3361 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
3421 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
3481 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
3541 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
3601 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
3661 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
3721 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
3781 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
3841 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
3901 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
3961 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
4021 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
4081 aaaaaggatc tcaaagctgt atcgaagatc ctttgatctt ttctacgggg tctgacgctc
4141 agtggaacga aaactcacgt taagggattt tggtcatgaa caataaaact gtctgcttac
4201 ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc ttgctctagg
4261 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
4321 gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc gccagagttg
4381 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
4441 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
4501 gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt attagaagaa
4561 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
4621 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
4681 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
4741 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
4801 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
4861 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
4921 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
4981 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
5041 gaattaattc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
5101 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
5161 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
5221 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
5281 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
5341 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
5401 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
5461 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
5521 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
5581 ggcgcgtccc attcgcca
//