質(zhì)粒序列
LOCUS Exported 6346 bp ds-DNA circular SYN 13-8-2015
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6346)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-8-13 from MLCC
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..6346
/organism="synthetic DNA construct"
/mol_type="other DNA"
source 1283..1305
/organism="Enterobacteria phage T7"
/mol_type="genomic DNA"
/db_xref="taxon:10760"
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(527..547)
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/note="TEV site"
/translation="ENLYFQG"
CDS complement(584..1234)
/codon_start=1
/product="glutathione S-transferase from Schistosoma
japonicum"
/note="GST"
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPP"
CDS complement(1253..1270)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
RBS 1283..1305
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 1320..1344
/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(1345..1363)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 1672..1749
/gene="lacI"
/note="lacI promoter"
CDS 1750..2832
/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 3641..3832
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 3934..4076
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(4262..4850)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 4972..5787
/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 complement(5880..6335)
/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 tccggtacct atgtcataga cacggcaact tgttcgatga
241 agctggccag gtttatatcc cgttcagaaa gaccggcaca ttcgtgggtg ctcaaggtga
301 tatggacctt gttgtacacg ttaaaccact cgggatggtg gtccagcttt tcagcctgca
361 gggcgactct tgtcatgaag ccaaaagccc tgttgaagtc tttaaaatgg aactgtttga
421 agatggcatc tcggccttcc agttcattcc accccacggc ccgcaggttt ggcagcagct
481 ggtcccgttc ctcagcactc agcctgtgtg ccttgccagc catggcgccc tgaaaataaa
541 gattctcgct catccatccg ccaccaccac cagatccact agttggagga tggtcgccac
601 caccaaacgt ggcttgccag ccctgcaaag gccatgctat atacttgctg gatttcaagt
661 acttatcaat ttgtgggata gcttcaatac gttttttaaa acaaactaat tttgggaacg
721 catccaggca cattgggtcc atgtataaaa caacatcaag agcgtcatac aacatgaagt
781 caggatgggt tacatgatca ccatttaaat atgttttatg acataaacga tcttcgaaca
841 ttttcagcat ttcaggtagc ttgctaagaa aatcaacttt gagagtttca aagtctttac
901 tatatgcaat tctcgaaaca ccgtatctaa tatccaaaac cgctccttca agcattgaaa
961 tctctgcacg ctcttttgga caaccaccca acatgttgtg cttgtcagct atataacgta
1021 tgatggccat agactgtgtt aatttaacat caccatcaat ataataagga agattgggaa
1081 actccaaacc caattcaaac tttttgtttc gccatttatc accttcatcg cgctcataca
1141 aatgctcttc atatttttct tcaagatatt ccaaaagaag tcgagtgggt tgcacaaggc
1201 ccttaatttt ccaataacct agtatagggg acatggaatt gctactagtg ttgtgatggt
1261 gatggtgatg tttcatggta tatctccttc ttaaagttaa acaaaattat ttctagaggg
1321 gaattgttat ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc gggatcgaga
1381 tctcgatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg
1441 ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca
1501 tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg
1561 ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac
1621 tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgagatccc ggacaccatc
1681 gaatggcgca aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg
1741 gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat
1801 cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa
1861 gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg
1921 ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg
1981 caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg
2041 atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa
2101 cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa
2161 gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac
2221 agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg
2281 ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt
2341 ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa
2401 ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc
2461 gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt
2521 accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa
2581 gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg
2641 caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag
2701 ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc
2761 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
2821 agcgggcagt gagcgcaacg caattaatgt aagttagctc actcattagg caccgggatc
2881 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
2941 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
3001 ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg
3061 cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc
3121 cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccccacgggt
3181 gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg
3241 ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc
3301 tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac
3361 gcggaagtca gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct
3421 accctgtgga acacctacat ctgtattaac gaagcgctgg cattgaccct gagtgatttt
3481 tctctggtcc cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg
3541 ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat
3601 tacccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac
3661 cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa
3721 cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga
3781 gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
3841 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
3901 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
3961 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
4021 catatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
4081 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
4141 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
4201 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
4261 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
4321 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
4381 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
4441 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
4501 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
4561 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
4621 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
4681 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
4741 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
4801 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
4861 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
4921 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
4981 attcaacggg aaacgtcttg ctctaggccg cgattaaatt ccaacatgga tgctgattta
5041 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg
5101 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
5161 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
5221 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccggg
5281 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
5341 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
5401 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
5461 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
5521 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
5581 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
5641 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
5701 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
5761 catttgatgc tcgatgagtt tttctaagaa ttaattcatg agcggataca tatttgaatg
5821 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
5881 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
5941 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
6001 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
6061 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
6121 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
6181 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
6241 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
6301 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgcca
//