質(zhì)粒序列
LOCUS Exported 7311 bp ds-DNA circular SYN 11-8-2015
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7311)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-8-11 from MLCC
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..7311
/organism="synthetic DNA construct"
/mol_type="other DNA"
source 2362..2384
/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"
terminator 141..168
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 260..346
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(512..529)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(536..568)
/codon_start=1
/product="HSV (herpes simplex virus) epitope tag"
/note="HSV tag"
/translation="QPELAPEDPED"
CDS complement(681..695)
/codon_start=1
/product="enterokinase recognition and cleavage site"
/note="enterokinase site"
/translation="DDDDK"
CDS complement(714..731)
/codon_start=1
/product="thrombin recognition and cleavage site"
/note="thrombin site"
/translation="LVPRGS"
CDS complement(747..791)
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/note="S-Tag"
/translation="KETAAAKFERQHMDS"
CDS complement(801..818)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(834..2318)
/codon_start=1
/gene="nusA"
/product="transcription elongation protein (N utilization
substance protein A)"
/note="NusA"
/note="highly soluble in E. coli"
/translation="MNKEILAVVEAVSNEKALPREKIFEALESALATATKKKYEQEIDV
RVQIDRKSGDFDTFRRWLVVDEVTQPTKEITLEAARYEDESLNLGDYVEDQIESVTFDR
ITTQTAKQVIVQKVREAERAMVVDQFREHEGEIITGVVKKVNRDNISLDLGNNAEAVIL
REDMLPRENFRPGDRVRGVLYSVRPEARGAQLFVTRSKPEMLIELFRIEVPEIGEEVIE
IKAAARDPGSRAKIAVKTNDKRIDPVGACVGMRGARVQAVSTELGGERIDIVLWDDNPA
QFVINAMAPADVASIVVDEDKHTMDIAVEAGNLAQAIGRNGQNVRLASQLSGWELNVMT
VDDLQAKHQAEAHAAIDTFTKYLDIDEDFATVLVEEGFSTLEELAYVPMKELLEIEGLD
EPTVEALRERAKNALATIAQAQEESLGDNKPADDLLNLEGVDRDLAFKLAARGVCTLED
LAEQGIDDLADIEGLTDEKAGALIMAARNICWFGDEA"
CDS complement(2325..2342)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
RBS 2362..2384
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 2399..2423
/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(2424..2442)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 2755..2832
/gene="lacI"
/note="lacI promoter"
CDS 2833..3915
/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 4487..4678
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 4780..4922
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(5108..5696)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 5777..5869
/gene="bla"
/note="AmpR promoter"
CDS 5870..6730
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin complement(6845..7300)
/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 tcagcggata catatttgaa tgtatttaga aaaataaaca
121 aaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc ttaatttgat
181 gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt
241 tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga
301 taaaacgaaa ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggcagttc
361 cctactctcg catggggaga ccccacacta ccatcggcgc tacggcgttt cacttctgag
421 ttcggcatgg ggtcaggtgg gaccaccgcg ctactgccgc caggcaaatt ctgttttatc
481 agcctaggaa cgcccaactt aattaacatt agtggtggtg gtggtggtgc tcgagatcct
541 ctgggtcttc aggagcgagt tctggctggc ttgcacgtgt atacagctgt gcggccgcaa
601 gcttcgaacg cgtatcgatg gtaccgtcga cgtcctgcag gcgcgcctgt acagaattcg
661 gatccacgag ctcccggact cttgtcgtcg tcatcaatcg taccagaacc cgcgctgccc
721 cgggggacca gaccagttgg cggtggcgag tccatgtgct ggcgttcaaa tttcgcagca
781 gcggtttctt tacccgcgga gtgatggtga tggtgatgac cagaaccact agtcgcttcg
841 tcaccgaacc agcaaatatt acgggcagcc ataatcagtg ctccggcttt ttcgtcggtc
901 aacccttcga tatcagccag atcatcaatg ccctgttcgg cgagatcttc cagcgtacaa
961 acgccacggg cggccagttt gaatgccaaa tcacgatcta ccccttcaag gttcagcaga
1021 tcgtcagccg gtttgttatc accgaggctt tcttcctggg cctgtgcaat ggtggccagt
1081 gcatttttag cacgctcgcg cagtgcttca acggtcggct catcaaggcc ttcgatttcc
1141 aacagctctt tcatcggcac ataggccaat tcttccagcg tcgagaagcc ttcttctacc
1201 agaacagtcg cgaagtcttc gtcgatgtcg agatatttgg tgaaggtgtc gatcgctgcg
1261 tgcgcttccg cctgatgctt agcttgcagg tcgtcaacgg tcatcacgtt gagttcccaa
1321 ccgctcagtt gcgaagccag acgcacgttc tgaccgttac ggccaatcgc ctgcgccaga
1381 ttaccggctt caacggcgat gtccatggtg tgtttatctt catccaccac gatagaagca
1441 acgtctgccg gtgccattgc gttaatcacg aactgcgccg ggttatcatc ccacaggacg
1501 atatcgatac gctcgccacc cagttcagta gacaccgcct gaacacgcgc gccacgcata
1561 cctacgcaag cacctaccgg atcgatacgt ttatcgttgg ttttcaccgc gattttcgca
1621 cgagaacccg gatcgcgagc cgctgcttta atttcaatca cttcttcgcc gatttctggc
1681 acttcaatac ggaacagttc gatcagcatt tccggcttgg aacgagtgac gaacagttgc
1741 gcgccacgcg cttccgggcg aacggaatag agcacgccac gaacgcggtc gccagggcgg
1801 aagttttcac gcggcagcat atcttcgcgc aggatcacgg cttcagcgtt gttgcccaga
1861 tccagagaga tgttgtcgcg gtttactttt ttcaccacgc cggtgatgat ttcaccttcg
1921 tgttcacgga actgatcaac caccatcgca cgttcggctt cacgcacttt ctgcacgata
1981 acctgttttg ccgtctgggt agtgatacgg tcaaaggtaa cagactcaat ctgatcttca
2041 acgtaatcgc ccaggttcag gctttcatct tcataacgtg cggcttcaag ggtgatttcc
2101 ttggtcggct gggtgacttc atcaacaact aaccagcgac ggaaagtgtc aaaatcaccg
2161 cttttgcgat cgatctgtac gcggacgtcg atctcttgtt catatttttt ctttgttgct
2221 gtcgccagcg cgctttccaa tgcttcgaaa atcttctcgc gaggtagcgc cttttcattg
2281 gatacggctt caactacagc caaaatttct ttgttcatac tagaatggtg atggtggtga
2341 tggctgctgc ccatatgtat atctccttct taaagttaaa caaaattatt tctagagggg
2401 aattgttatc cgctcacaat tcccctatag tgagtcgtat taatttcgcg ggatcgagat
2461 cgatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg
2521 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
2581 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
2641 gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac
2701 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat cccggacacc
2761 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
2821 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
2881 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
2941 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
3001 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
3061 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
3121 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
3181 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
3241 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
3301 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
3361 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
3421 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
3481 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
3541 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
3601 attaccgagt ccgggctgcg cgttggtgcg gacatctcgg tagtgggata cgacgatacc
3661 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
3721 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
3781 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
3841 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
3901 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcaccggg
3961 atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
4021 catgactagc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct
4081 tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa
4141 aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct
4201 ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg atctgcatcg caggatgctg
4261 ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt
4321 gatttttctc tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag
4381 taaccgggca tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg
4441 tatcattacc cccatgaaca gaaatccccc ttacacggag gcatcagtga ccaaacagga
4501 aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa
4561 actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc
4621 tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
4681 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
4741 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
4801 tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga
4861 gtgcaccata tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
4921 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
4981 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
5041 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
5101 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
5161 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
5221 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
5281 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
5341 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
5401 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
5461 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
5521 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
5581 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
5641 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
5701 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
5761 ttttggtcat gacctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
5821 agacaataac cctgataaat gcttcaatca tgattgaaaa aggaagagta tgagtattca
5881 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
5941 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
6001 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
6061 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
6121 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
6181 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
6241 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
6301 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
6361 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
6421 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
6481 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
6541 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
6601 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
6661 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
6721 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
6781 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatgc catcgtgccg
6841 ccagaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
6901 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
6961 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
7021 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
7081 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
7141 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
7201 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
7261 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc a
//