質(zhì)粒序列
LOCUS Exported 4414 bp ds-DNA circular SYN 29-七月-2016
DEFINITION synthetic circular DNA
KEYWORDS pTrcHisA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4414)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2016年7月29日 from SnapGene Viewer 2.8.1
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..4414
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 193..222
/note="trc promoter"
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 230..246
/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)."
misc_feature 263..332
/note="rrnG antiterminator"
/note="antiterminator from the E. coli rrnG leader region
(Berg et al., 1989)"
CDS 383..409
/codon_start=1
/product="synthetic cistron containing a ribosome binding
site (Shine-Dalgarno sequence), for enhancing the bacterial
expression of a downstream cistron (Schoner, 1997)"
/note="minicistron"
/note="This first cistron should terminate 3 bp upstream of
the ATG for the second cistron."
/translation="MYRLNKEE"
CDS 425..442
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
CDS 446..478
/codon_start=1
/product="leader peptide from bacteriophage T7 gene 10"
/note="T7 tag (gene 10 leader)"
/note="promotes efficient translation in E. coli"
/translation="MASMTGGQQMG"
CDS 482..505
/codon_start=1
/product="Xpress(TM) epitope tag, including an enterokinase
recognition and cleavage site"
/note="Xpress(TM) tag"
/translation="DLYDDDDK"
misc_feature 515..564
/note="MCS"
terminator 767..853
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 945..972
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 992..1083
/gene="bla"
/note="AmpR promoter"
CDS 1084..1944
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 2115..2703
/direction=RIGHT
/note="ColE1 ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 2889..3029
/note="bom"
/note="basis of mobility region from pBR322"
promoter 3215..3292
/gene="lacI"
/note="lacI promoter"
CDS 3293..4375
/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
YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
ORIGIN
1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
241 taacaatttc acacaggaaa cagcgccgct gagaaaaagc gaagcggcac tgctctttaa
301 caatttatca gacaatctgt gtgggcactc gaccggaatt atcgattaac tttattatta
361 aaaattaaag aggtatatat taatgtatcg attaaataag gaggaataaa ccatgggggg
421 ttctcatcat catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg
481 ggatctgtac gacgatgacg ataaggatcg atggggatcc gagctcgaga tctgcagctg
541 gtaccatatg ggaattcgaa gcttggctgt tttggcggat gagagaagat tttcagcctg
601 atacagatta aatcagaacg cagaagcggt ctgataaaac agaatttgcc tggcggcagt
661 agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat
721 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa
781 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
841 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg
901 gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac
961 ggatggcctt tttgcgtttc tacaaactct ttttgtttat ttttctaaat acattcaaat
1021 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
1081 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
1141 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
1201 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
1261 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
1321 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
1381 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
1441 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
1501 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
1561 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
1621 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
1681 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
1741 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
1801 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
1861 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
1921 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
1981 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
2041 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
2101 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
2161 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
2221 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
2281 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
2341 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
2401 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
2461 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
2521 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
2581 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
2641 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
2701 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
2761 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
2821 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
2881 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
2941 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
3001 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
3061 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
3121 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag cagatcaatt
3181 cgcgcgcgaa ggcgaagcgg catgcattta cgttgacacc atcgaatggc gcaaaacctt
3241 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
3301 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
3361 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
3421 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
3481 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
3541 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
3601 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
3661 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
3721 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
3781 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
3841 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
3901 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
3961 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
4021 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
4081 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
4141 cccgccgtca accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
4201 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
4261 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
4321 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
4381 acgcaattaa tgtgagttag cgcgaattga tctg
//