質(zhì)粒序列
LOCUS Exported 5372 bp ds-DNA circular SYN 05-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 5
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5372)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Monday, September 5, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..5372
/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(217..234)
/codon_start=1
/product="thrombin recognition and cleavage site"
/note="thrombin site"
/translation="LVPRGS"
CDS complement(250..294)
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/note="S-Tag"
/translation="KETAAAKFERQHMDS"
RBS 305..327
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 342..366
/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(367..385)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 698..775
/gene="lacI"
/note="lacI promoter"
CDS 776..1858
/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 2667..2858
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 2960..3102
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(3288..3876)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 3998..4813
/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(4906..5361)
/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 tccacagata tcccatggaa ccgcgtggca ccagggtacc
241 cagatctggg ctgtccatgt gctggcgttc gaatttagca gcagcggttt ctttcatatg
301 tatatctcct tcttaaagtt aaacaaaatt atttctagag gggaattgtt atccgctcac
361 aattccccta tagtgagtcg tattaatttc gcgggatcga gatcgatctc gatcctctac
421 gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc
481 gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc
541 ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat
601 gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta
661 atgcaggagt cgcataaggg agagcgtcga gatcccggac accatcgaat ggcgcaaaac
721 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa
781 accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg
841 cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat
901 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt
961 gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc
1021 gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag
1081 cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct
1141 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa
1201 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc
1261 ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat
1321 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca
1381 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
1441 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
1501 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
1561 gcgcgttggt gcggacatct cggtagtggg atacgacgat accgaagaca gctcatgtta
1621 tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
1681 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
1741 actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt
1801 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
1861 gcaacgcaat taatgtaagt tagctcactc attaggcacc gggatctcga ccgatgccct
1921 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg
1981 cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg
2041 tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg
2101 tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt
2161 tcggcgagaa gcaggccatt atcgccggca tggcggcccc acgggtgcgc atgatcgtgc
2221 tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct tactggttag cagaatgaat
2281 caccgatacg cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa
2341 caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc
2401 cctgcaccat tatgttccgg atctgcatcg caggatgctg ctggctaccc tgtggaacac
2461 ctacatctgt attaacgaag cgctggcatt gaccctgagt gatttttctc tggtcccgcc
2521 gcatccatac cgccagttgt ttaccctcac aacgttccag taaccgggca tgttcatcat
2581 cagtaacccg tatcgtgagc atcctctctc gtttcatcgg tatcattacc cccatgaaca
2641 gaaatccccc ttacacggag gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg
2701 cccgctttat cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg
2761 atgaacaggc agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct
2821 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
2881 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
2941 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata
3001 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tatgcggtgt
3061 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg
3121 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
3181 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
3241 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
3301 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
3361 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
3421 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
3481 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
3541 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
3601 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
3661 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
3721 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
3781 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
3841 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
3901 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gaacaataaa
3961 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
4021 gtcttgctct aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
4081 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga
4141 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
4201 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
4261 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccgggaaaa cagcattcca
4321 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
4381 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
4441 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
4501 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
4561 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
4621 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
4681 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
4741 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
4801 tgagtttttc taagaattaa ttcatgagcg gatacatatt tgaatgtatt tagaaaaata
4861 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgaaatt gtaaacgtta
4921 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
4981 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
5041 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
5101 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
5161 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
5221 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
5281 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
5341 atgcgccgct acagggcgcg tcccattcgc ca
//