質(zhì)粒序列
LOCUS Exported File 4968 bp ds-DNA circular SYN 28-1-2015
DEFINITION .
ACCESSION .
VERSION .
KEYWORDS pGEX4T-3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4968)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported 2015-1-28 from SnapGene 2.0.1
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..4968
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 183..211
/note="tac promoter"
/note="strong E. coli promoter; hybrid between the trp and
lac?UV5 promoters"
protein_bind 219..235
/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)."
CDS 258..911
/codon_start=1
/gene="
"
/product="glutathione S-transferase from Schistosoma
japonicum"
/note="GST"
/translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
CDS 918..935
/codon_start=1
/product="thrombin recognition and cleavage site"
/note="thrombin site"
/translation="LVPRGS"
promoter 1271..1375
/gene="bla"
/note="AmpR promoter"
CDS 1376..2236
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 2407..2995
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 3239..3316
/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."
CDS 3317..4399
/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"
protein_bind 4412..4433
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4448..4478
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 4486..4502
/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)."
primer_bind 4510..4526
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4542..4558)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
901 atcctccaaa atcggatctg gttccgcgtg gatccccgaa ttcccgggtc gactcgagcg
961 gccgcatcgt gactgactga cgatctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
1021 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
1081 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
1141 gtcacgtagc gatagcggag tgtataattc ttgaagacga aagggcctcg tgatacgcct
1201 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
1261 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
1321 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
1381 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
1441 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
1501 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
1561 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
1621 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
1681 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
1741 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
1801 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
1861 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
1921 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
1981 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
2041 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
2101 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
2161 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
2221 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
2281 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
2341 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
2401 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
2461 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
2521 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
2581 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
2641 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
2701 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
2761 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
2821 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
2881 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
2941 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
3001 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
3061 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
3121 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
3181 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca taaattccga
3241 caccatcgaa tggtgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
3301 attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt
3361 ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg
3421 ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca
3481 actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc
3541 gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt
3601 ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct
3661 cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc
3721 tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc
3781 catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt
3841 cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg
3901 tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga
3961 acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga
4021 gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg
4081 cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga
4141 taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct
4201 gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg
4261 caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca
4321 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
4381 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
4441 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
4501 aatttcacac aggaaacagc tatgaccatg attacggatt cactggccgt cgttttacaa
4561 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
4621 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
4681 agcctgaatg gcgaatggcg ctttgcctgg tttccggcac cagaagcggt gccggaaagc
4741 tggctggagt gcgatcttcc tgaggccgat actgtcgtcg tcccctcaaa ctggcagatg
4801 cacggttacg atgcgcccat ctacaccaac gtaacctatc ccattacggt caatccgccg
4861 tttgttccca cggagaatcc gacgggttgt tactcgctca catttaatgt tgatgaaagc
4921 tggctacagg aaggccagac gcgaattatt tttgatggcg ttggaatt
//