质粒载体
pEX18GM大肠自杀质粒
点击下载附件附件
| 复制子: | pBR322 |
|---|---|
| 原核抗性: | Gen |
| 筛选标记: | SacB |
| 克隆菌株: | DH5a |
| 培养条件: | 37度 |
| 质粒宿主： | 大肠杆菌 |
|---|---|
| 质粒用途： | 基因编辑 |
| 片段类型： | |
| 片段物种： | |
| 原核抗性： | Gen |
| 真核抗性： | |
| 荧光标记： |
pEX18GM是一个大肠杆菌SacB自杀质粒。
LOCUS Exported 5831 bp ds-DNA circular SYN 08-JAN-2018
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..5831
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(209..1630)
/codon_start=1
/gene="Bacillus subtilis sacB"
/product="secreted levansucrase that renders bacterial
growth sensitive to sucrose"
/label=SacB
/note="negative selection marker"
/translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
VKDSILEQGQLTVNK"
promoter complement(1631..2076)
/gene="sacR"
/label=sacB promoter
/note="sacB promoter and control region"
oriT 2450..2559
/note="incP origin of transfer"
terminator 2980..3007
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 3099..3185
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(3341..3625)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRAWLFWRMREDFQPDTD"
misc_feature 3555..3611
/label=MCS
/note="pUC18/19 multiple cloning site"
protein_bind 3645..3661
/label=lac operator
/bound_moiety="lac repressor encoded by 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)."
promoter complement(3669..3699)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3714..3735
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4023..4611)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4864..5397)
/codon_start=1
/gene="aacC1"
/product="gentamycin acetyltransferase"
/label=Gen
/note="confers resistance to gentamycin"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(5586..5614)
/gene="intI1 (promoter lies within the coding sequence)"
/label=Pc promoter
/note="class 1 integron promoter"
promoter complement(5632..5736)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
181 accataatcg gcattttctt ttgcgttttt atttgttaac tgttaattgt ccttgttcaa
241 ggatgctgtc tttgacaaca gatgttttct tgcctttgat gttcagcagg aagctaggcg
301 caaacgttga ttgtttgtct gcgtagaatc ctctgtttgt catatagctt gtaatcacga
361 cattgtttcc tttcgcttga ggtacagcga agtgtgagta agtaaaggtt acatcgttag
421 gatcaagatc catttttaac acaaggccag ttttgttcag cggcttgtat gggccagtta
481 aagaattaga aacataacca agcatgtaaa tatcgttaga cgtaatgccg tcaatcgtca
541 tttttgatcc gcgggagtca gtgaacagat accatttgcc gttcatttta aagacgttcg
601 cgcgttcaat ttcatctgtt actgtgttag atgcaatcag cggtttcatc acttttttca
661 gtgtgtaatc atcgtttagc tcaatcatac cgagagcgcc gtttgctaac tcagccgtgc
721 gttttttatc gctttgcaga agtttttgac tttcttgacg gaagaatgat gtgcttttgc
781 catagtatgc tttgttaaat aaagattctt cgccttggta gccatcttca gttccagtgt
841 ttgcttcaaa tactaagtat ttgtggcctt tatcttctac gtagtgagga tctctcagcg
901 tatggttgtc gcctgagctg tagttgcctt catcgatgaa ctgctgtaca ttttgatacg
961 tttttccgtc accgtcaaag attgatttat aatcctctac accgttgatg ttcaaagagc
1021 tgtctgatgc tgatacgtta acttgtgcag ttgtcagtgt ttgtttgccg taatgtttac
1081 cggagaaatc agtgtagaat aaacggattt ttccgtcaga tgtaaatgtg gctgaacctg
1141 accattcttg tgtttggtct tttaggatag aatcatttgc atcgaatttg tcgctgtctt
1201 taaagacgcg gccagcgttt ttccagctgt caatagaagt ttcgccgact ttttgataga
1261 acatgtaaat cgatgtgtca tccgcatttt taggatctcc ggctaatgca aagacgatgt
1321 ggtagccgtg atagtttgcg acagtgccgt cagcgttttg taatggccag ctgtcccaaa
1381 cgtccaggcc ttttgcagaa gagatatttt taattgtgga cgaatcgaac tcaggaactt
1441 gatatttttc atttttttgc tgttcaggga tttgcagcat atcatggcgt gtaatatggg
1501 aaatgccgta tgtttcctta tatggctttt ggttcgtttc tttcgcaaac gcttgagttg
1561 cgcctcctgc cagcagtgcg gtagtaaagg ttaatactgt tgcttgtttt gcaaactttt
1621 tgatgttcat cgttcatgtc tcctttttta tgtactgtgt tagcggtctg cttcttccag
1681 ccctcctgtt tgaagatggc aagttagtta cgcacaataa aaaaagacct aaaatatgta
1741 aggggtgacg ccaaagtata cactttgccc tttacacatt ttaggtcttg cctgctttat
1801 cagtaacaaa cccgcgcgat ttacttttcg acctcattct attagactct cgtttggatt
1861 gcaactggtc tattttcctc ttttgtttga tagaaaatca taaaaggatt tgcagactac
1921 gggcctaaag aactaaaaaa tctatctgtt tcttttcatt ctctgtattt tttatagttt
1981 ctgttgcatg ggcataaagt tgccttttta atcacaattc agaaaatatc ataatatctc
2041 atttcactaa ataatagtga acggcaggta tatgtgatgg gttaaaaagg atcgatcctc
2101 tagctagagt cgatcttcgc cagcagggcg aggatcgtgg catcaccgaa ccgcgccgtg
2161 cgcgggtcgt cggtgagcca gagtttcagc aggccgccca ggcggcccag gtcgccattg
2221 atgcgggcca gctcgcggac gtgctcatag tccacgacgc ccgtgatttt gtagccctgg
2281 ccgacggcca gcaggtaggc cgacaggctc atgccggccg ccgccgcctt ttcctcaatc
2341 gctcttcgtt cgtctggaag gcagtacacc ttgataggtg ggctgccctt cctggttggc
2401 ttggtttcat cagccatccg cttgccctca tctgttacgc cggcggtagc cggccagcct
2461 cgcagagcag gattcccgtt gagcaccgcc aggtgcgaat aagggacagt gaagaaggaa
2521 cacccgctcg cgggtgggcc tacttcacct atcctgcccg gctgacgccg ttggatacac
2581 caaggaaagt ctacacgaac cctttggcaa aatcctgtat atcgtgcgaa aaaggatgga
2641 tataccgaaa aaatcgctat aatgaccccg aagcagggtt atgcagcgga aaagcgctgc
2701 ttccctgctg ttttgtggaa tatctaccga ctggaaacag gcaaatgcag gaaattactg
2761 aactgagggg acaggcgaga gacgatgcca aagagctaca ccgacgagct ggccgagtgg
2821 gttgaatccc gcgcggccaa gaagcgccgg cgtgatgagg ctgcggttgc gttcctggcg
2881 gtgagggcgg atgtcgatat gcgtaaggag aaaataccgc atcaggcgca tatttgaatg
2941 tatttagaaa aataaacaaa aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg
3001 gccttctgct taatttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg
3061 ggccgttgct tcgcaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg
3121 ttcaccgaca aacaacagat aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt
3181 atttgatgcc tggcagttcc ctactctcgc atggggagac cccacactac catcggcgct
3241 acggcgtttc acttctgagt tcggcatggg gtcaggtggg accaccgcgc tactgccgcc
3301 aggcaaattc tgttttatca gaccgcttct gcgttctgat ttaatctgta tcaggctgaa
3361 aatcttctct catccgccaa aacagccaag ctcgccattc gccattcagg ctgcgcaact
3421 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
3481 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
3541 cgacggccag tgccaagctt gcatgcctgc aggtcgactc tagaggatcc ccgggtaccg
3601 agctcgaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
3661 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
3721 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
3781 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
3841 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
3901 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
3961 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
4021 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
4081 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
4141 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
4201 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
4261 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
4321 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
4381 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
4441 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
4501 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
4561 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
4621 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
4681 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
4741 aaatcaatct aaagtatata tgagtaaact tggtctgaca atcgatgcga attggccgcg
4801 gcgttgtgac aatttaccga acaactccgc ggccgggaag ccgatctcgg cttgaacgaa
4861 ttgttaggtg gcggtacttg ggtcgatatc aaagtgcatc acttcttccc gtatgcccaa
4921 ctttgtatag agagccactg cgggatcgtc accgtaatct gcttgcacgt agatcacata
4981 agcaccaagc gcgttggcct catgcttgag gagattgatg agcgcggtgg caatgccctg
5041 cctccggtgc tcgccggaga ctgcgagatc atagatatag atctcactac gcggctgctc
5101 aaacctgggc agaacgtaag ccgcgagagc gccaacaacc gcttcttggt cgaaggcagc
5161 aagcgcgatg aatgtcttac tacggagcaa gttcccgagg taatcggagt ccggctgatg
5221 ttgggagtag gtggctacgt ctccgaactc acgaccgaaa agatcaagag cagcccgcat
5281 ggatttgact tggtcagggc cgagcctaca tgtgcgaatg atgcccatac ttgagccacc
5341 taactttgtt ttagggcgac tgccctgctg cgtaacatcg ttgctgctgc gtaacatcgt
5401 tgctgctcca taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc
5461 gaggcataga ctgtacaaaa aaacagtcat aacaagccat gaaaaccgcc actgcgccgt
5521 taccaccgct gcgttcggtc aaggttctgg accagttgcg tgagcgcata cgctacttgc
5581 attacagttt acgaaccgaa caggcttatg tcaattcggt tgaatactca tactcttcct
5641 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
5701 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
5761 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
5821 gccctttcgt c