质粒序列
LOCUS Exported 7273 bp ds-DNA circular SYN 06-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7273)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Tuesday, September 6, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..7273
/organism="synthetic DNA construct"
/mol_type="other DNA"
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(679..693)
/codon_start=1
/product="enterokinase recognition and cleavage site"
/note="enterokinase site"
/translation="DDDDK"
CDS complement(712..729)
/codon_start=1
/product="thrombin recognition and cleavage site"
/note="thrombin site"
/translation="LVPRGS"
CDS complement(745..789)
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/note="S-Tag"
/translation="KETAAAKFERQHMDS"
CDS complement(799..816)
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS complement(832..2316)
/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"
RBS 2324..2346
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
protein_bind 2361..2385
/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(2386..2404)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter 2717..2794
/gene="lacI"
/note="lacI promoter"
CDS 2795..3877
/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 4449..4640
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/note="rop"
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
misc_feature 4742..4884
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(5070..5658)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 5739..5831
/gene="bla"
/note="AmpR promoter"
CDS 5832..6692
/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(6807..7262)
/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 ctcggactct tgtcgtcgtc atcaatcgta ccagaacccg cgctgccccg
721 ggggaccaga ccagttggcg gtggcgagtc catgtgctgg cgttcaaatt tcgcagcagc
781 ggtttcttta cccgcggagt gatggtgatg gtgatgacca gaaccactag tcgcttcgtc
841 accgaaccag caaatattac gggcagccat aatcagtgct ccggcttttt cgtcggtcaa
901 cccttcgata tcagccagat catcaatgcc ctgttcggcg agatcttcca gcgtacaaac
961 gccacgggcg gccagtttga atgccaaatc acgatctacc ccttcaaggt tcagcagatc
1021 gtcagccggt ttgttatcac cgaggctttc ttcctgggcc tgtgcaatgg tggccagtgc
1081 atttttagca cgctcgcgca gtgcttcaac ggtcggctca tcaaggcctt cgatttccaa
1141 cagctctttc atcggcacat aggccaattc ttccagcgtc gagaagcctt cttctaccag
1201 aacagtcgcg aagtcttcgt cgatgtcgag atatttggtg aaggtgtcga tcgctgcgtg
1261 cgcttccgcc tgatgcttag cttgcaggtc gtcaacggtc atcacgttga gttcccaacc
1321 gctcagttgc gaagccagac gcacgttctg accgttacgg ccaatcgcct gcgccagatt
1381 accggcttca acggcgatgt ccatggtgtg tttatcttca tccaccacga tagaagcaac
1441 gtctgccggt gccattgcgt taatcacgaa ctgcgccggg ttatcatccc acaggacgat
1501 atcgatacgc tcgccaccca gttcagtaga caccgcctga acacgcgcgc cacgcatacc
1561 tacgcaagca cctaccggat cgatacgttt atcgttggtt ttcaccgcga ttttcgcacg
1621 agaacccgga tcgcgagccg ctgctttaat ttcaatcact tcttcgccga tttctggcac
1681 ttcaatacgg aacagttcga tcagcatttc cggcttggaa cgagtgacga acagttgcgc
1741 gccacgcgct tccgggcgaa cggaatagag cacgccacga acgcggtcgc cagggcggaa
1801 gttttcacgc ggcagcatat cttcgcgcag gatcacggct tcagcgttgt tgcccagatc
1861 cagagagatg ttgtcgcggt ttactttttt caccacgccg gtgatgattt caccttcgtg
1921 ttcacggaac tgatcaacca ccatcgcacg ttcggcttca cgcactttct gcacgataac
1981 ctgttttgcc gtctgggtag tgatacggtc aaaggtaaca gactcaatct gatcttcaac
2041 gtaatcgccc aggttcaggc tttcatcttc ataacgtgcg gcttcaaggg tgatttcctt
2101 ggtcggctgg gtgacttcat caacaactaa ccagcgacgg aaagtgtcaa aatcaccgct
2161 tttgcgatcg atctgtacgc ggacgtcgat ctcttgttca tattttttct ttgttgctgt
2221 cgccagcgcg ctttccaatg cttcgaaaat cttctcgcga ggtagcgcct tttcattgga
2281 tacggcttca actacagcca aaatttcttt gttcatatgt atatctcctt cttaaagtta
2341 aacaaaatta tttctagagg ggaattgtta tccgctcaca attcccctat agtgagtcgt
2401 attaatttcg cgggatcgag atcgatctcg atcctctacg ccggacgcat cgtggccggc
2461 atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa
2521 gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc
2581 cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct tgcggcggcg
2641 gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga
2701 gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata
2761 gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt
2821 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
2881 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
2941 caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc
3001 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
3061 actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc
3121 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
3181 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
3241 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact
3301 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
3361 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca
3421 aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac
3481 catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat
3541 ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggacatctc
3601 ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat
3661 caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca
3721 gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac
3781 cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
3841 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt
3901 agctcactca ttaggcaccg ggatctcgac cgatgccctt gagagccttc aacccagtca
3961 gctccttccg gtgggcgcgg ggcatgacta gcatgatcgt gctcctgtcg ttgaggaccc
4021 ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata cgcgagcgaa
4081 cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga atggtcttcg
4141 gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc attatgttcc
4201 ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct gtattaacga
4261 agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat accgccagtt
4321 gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc cgtatcgtga
4381 gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc ccttacacgg
4441 aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc
4501 agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag gcagacatct
4561 gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg
4621 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
4681 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
4741 gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc
4801 atcagagcag attgtactga gagtgcacca tatatgcggt gtgaaatacc gcacagatgc
4861 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
4921 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
4981 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
5041 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
5101 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
5161 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
5221 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
5281 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
5341 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
5401 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
5461 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
5521 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
5581 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
5641 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
5701 aacgaaaact cacgttaagg gattttggtc atgacctatt tgtttatttt tctaaataca
5761 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat catgattgaa
5821 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
5881 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
5941 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
6001 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
6061 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
6121 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
6181 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
6241 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
6301 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
6361 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
6421 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
6481 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
6541 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
6601 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
6661 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
6721 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
6781 ataatctcat gccatcgtgc cgccagaaat tgtaaacgtt aatattttgt taaaattcgc
6841 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
6901 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
6961 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
7021 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
7081 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
7141 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
7201 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
7261 gtcccattcg cca
//