質(zhì)粒序列
LOCUS Exported 4753 bp ds-DNA circular SYN 08-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS PQE-81L
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4753)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Thursday, September 8, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..4753
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 10..54
/note="T5 promoter"
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 30..46
/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)."
protein_bind 62..78
/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)."
RBS 97..108
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 127..144
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
terminator 210..304
/note="lambda t0 terminator"
/note="transcription terminator from phage lambda"
CDS 348..1007
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/note="CmR"
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
terminator 1072..1158
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(1251..2333)
/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"
promoter complement(2334..2411)
/gene="lacI (mutant)"
/note="lacIq promoter"
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
misc_feature 2603..2743
/note="bom"
/note="basis of mobility region from pBR322"
rep_origin complement(2929..3517)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3688..4548)
/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"
promoter complement(4549..4653)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
121 ggatctcacc atcaccatca ccatacggat ccgcatgcga gctcggtacc ccgggtcgac
181 ctgcagccaa gcttaattag ctgagcttgg actcctgttg atagatccag taatgacctc
241 agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga
301 gaatccaagc tagcttggcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
361 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
421 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
481 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
541 gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag ctggtgatat
601 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc
661 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg
721 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg
781 tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca
841 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga
901 tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc
961 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca
1021 gttattggtg cccttaaacg cctggggtaa tgactctcta gcttgaggca tcaaataaaa
1081 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
1141 ctcctgagta ggacaaatcc gccctctaga ttacgtgcag tcgatgataa gctgtcaaac
1201 atgagaattg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc tcactgcccg
1261 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
1321 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
1381 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
1441 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
1501 tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag cccggactcg
1561 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
1621 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
1681 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
1741 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
1801 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
1861 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
1921 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
1981 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
2041 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
2101 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
2161 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
2221 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
2281 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
2341 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcaccat
2401 tcgatggtgt cggaatttcg ggcagcgttg ggtcctggcc acgggtgcgc atgatctaga
2461 gctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
2521 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
2581 cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt
2641 atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
2701 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc
2761 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
2821 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
2881 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
2941 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
3001 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
3061 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
3121 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
3181 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
3241 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
3301 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
3361 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
3421 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
3481 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
3541 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
3601 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
3661 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
3721 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
3781 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
3841 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
3901 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
3961 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
4021 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
4081 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
4141 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
4201 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
4261 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
4321 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
4381 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
4441 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
4501 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
4561 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
4621 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
4681 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
4741 ctttcgtctt cac
//