質(zhì)粒序列
LOCUS Exported 4412 bp ds-DNA circular SYN 11-9-2015
DEFINITION .
ACCESSION .
VERSION .
KEYWORDS Untitled 4
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4412)
AUTHORS admin
TITLE Direct Submission
JOURNAL Exported 2015-9-11 from MLCC
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..4412
/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"
terminator 765..851
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 943..970
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 990..1081
/gene="bla"
/note="AmpR promoter"
CDS 1082..1942
/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 2113..2701
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 2887..3027
/note="bom"
/note="basis of mobility region from pBR322"
promoter 3213..3290
/gene="lacI"
/note="lacI promoter"
/note="
"
CDS 3291..4373
/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 atggatccga cctcgagatc tgcagatggt
541 accatatggg aattcgaagc ttggctgttt tggcggatga gagaagattt tcagcctgat
601 acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg gcggcagtag
661 cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg
721 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
781 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
841 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc
901 gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg
961 atggcctttt tgcgtttcta caaactcttt ttgtttattt ttctaaatac attcaaatat
1021 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
1081 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
1141 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
1201 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
1261 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
1321 ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
1381 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
1441 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
1501 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
1561 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
1621 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
1681 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
1741 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
1801 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
1861 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
1921 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
1981 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
2041 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
2101 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
2161 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
2221 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
2281 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
2341 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
2401 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
2461 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
2521 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
2581 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
2641 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
2701 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
2761 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
2821 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
2881 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
2941 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
3001 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
3061 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
3121 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagca gatcaattcg
3181 cgcgcgaagg cgaagcggca tgcatttacg ttgacaccat cgaatggcgc aaaacctttc
3241 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
3301 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
3361 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
3421 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
3481 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
3541 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
3601 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
3661 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
3721 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
3781 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
3841 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
3901 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
3961 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
4021 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
4081 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
4141 cgccgtcaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
4201 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
4261 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
4321 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
4381 gcaattaatg tgagttagcg cgaattgatc tg
//