| 质粒名称: | pACM4 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 49797 |
| 存储实验室: | Mattheos Koffas |
|
载体骨架基本信息 |
|
| 载体名称: | pACYC-Duet |
| 载体出品公司: | Novagen |
| 载体抗性: | 氯霉素 |
| 空载体大小: | 5000 bp |
| 载体修饰: | AvrII was inserted upstream of the T7 promoter, XbaI was inserted between the operator (lacO) and ribosome binding site (RBS), SpeI was between the multiple-cloning site (MCS) and T7 terminator, and NheI was placed downstream of the T7 terminator |
| 载体类型: | 细菌表达 |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 启动子 | T7 |
| 标签/融合蛋白: | S-tag (C terminal on insert) |
| 克隆方法: | 限制性内切酶 |
|
文献引用方法 |
|
| 材料和方法部分: | pACM4 was a gift from Mattheos Koffas (爱迪基因 plasmid # 49797) |
| 参考文献部分: | ePathBrick: a synthetic biology platform for engineering metabolic pathways in E. coli. Xu P, Vansiri A, Bhan N, Koffas MA. ACS Synth Biol. 2012 Jul 20;1(7):256-66. doi: 10.1021/sb300016b. Epub 2012 May 4. 10.1021/sb300016b PubMed 23651248 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pACM4 3957 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION ePathBrick vector for pathway engineering in E. coli..
ACCESSION .
VERSION .
KEYWORDS pACM4
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3957)
AUTHORS Xu P, Vansiri A, Bhan N, Koffas MA
TITLE ePathBrick: a synthetic biology platform for engineering metabolic
pathways in E. coli.
JOURNAL ACS Synth Biol. 2012 Jul 20;1(7):256-66. doi: 10.1021/sb300016b.
Epub 2012 May 4.
PUBMED 23651248
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pACM4|
FEATURES Location/Qualifiers
source 1..3957
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 520..539
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(559..636)
/gene="lacI"
/label=lacI promoter
primer_bind 842..861
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 952..971
/label=T7
/note="T7 promoter, forward primer"
promoter 952..970
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 971..995
/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)."
RBS 1010..1032
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 1106..1150
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/label=S-Tag
/translation="KETAAAKFERQHMDS"
primer_bind complement(1189..1207)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 1203..1250
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(1376..2035)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 1950..1969
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(2036..2138)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
primer_bind complement(2530..2547)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2664..3208)
/direction=LEFT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
"
primer_bind complement(2744..2763)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3433..558)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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"
ORIGIN
1 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
61 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
121 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
181 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
241 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
301 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
361 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
421 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
481 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
541 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
601 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
661 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
721 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
781 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
841 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
901 cgatgcgtcc ggcgtagcct aggatcgaga tcgatctcga tcccgcgaaa ttaatacgac
961 tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact
1021 ttaagaagga gatatacata tggcagatct caattggata tcggccggcc acgcgatcgc
1081 tgacgtcggt accctcgagt ctggtaaaga aaccgctgct gcgaaatttg aacgccagca
1141 catggactcg tctactagtc gcagcttaat taacctaaac tgctgccacc gctgagcaat
1201 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctagcgaaag
1261 gaggagtcga ccgggtcgaa tttgctttcg aatttctgcc attcatccgc ttattatcac
1321 ttattcaggc gtagcaacca ggcgtttaag ggcaccaata actgccttaa aaaaattacg
1381 ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga
1441 agccatcaca gacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt
1501 gcgtataata tttgcccata gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt
1561 ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa
1621 taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata
1681 tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag
1741 tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt
1801 ctttcattgc catacggaac tccggatgag cattcatcag gcgggcaaga atgtgaataa
1861 aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca
1921 gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt
1981 tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag
2041 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
2101 cattatggtg aaagttggaa cctcttacgt gccgatcaac gtctcatttt cgccaaaagt
2161 tggcccaggg cttcccggta tcaacaggga caccaggatt tatttattct gcgaagtgat
2221 cttccgtcac aggtatttat tcggcgcaaa gtgcgtcggg tgatgctgcc aacttactga
2281 tttagtgtat gatggtgttt ttgaggtgct ccagtggctt ctgtttctat cagctgtccc
2341 tcctgttcag ctactgacgg ggtggtgcgt aacggcaaaa gcaccgccgg acatcagcgc
2401 aagcggagtg tatactggct tactatgttg gcactgatga gggtgtcagt gaagtgcttc
2461 atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag aatatgtgat acaggatata
2521 ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt cgactgcggc gagcggaaat
2581 ggcttacgaa cggggcggag atttcctgga agatgccagg aagatactta acagggaagt
2641 gagagggccg cggcaaagcc gtttttccat aggctccgcc cccctgacaa gcatcacgaa
2701 atctgacgct caaatcagtg gtggcgaaac ccgacaggac tataaagata ccaggcgttt
2761 cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc
2821 cgctgttatg gccgcgtttg tctcattcca cgcctgacac tcagttccgg gtaggcagtt
2881 cgctccaagc tggactgtat gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc
2941 ggtaactatc gtcttgagtc caacccggaa agacatgcaa aagcaccact ggcagcagcc
3001 actggtaatt gatttagagg agttagtctt gaagtcatgc gccggttaag gctaaactga
3061 aaggacaagt tttggtgact gcgctcctcc aagccagtta cctcggttca aagagttggt
3121 agctcagaga accttcgaaa aaccgccctg caaggcggtt ttttcgtttt cagagcaaga
3181 gattacgcgc agaccaaaac gatctcaaga agatcatctt attaatcaga taaaatattt
3241 ctagatttca gtgcaattta tctcttcaaa tgtagcacct gaagtcagcc ccatacgata
3301 taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc ggaaggagct gactgggttg
3361 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
3421 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
3481 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
3541 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
3601 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
3661 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatgtccgc
3721 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
3781 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
3841 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
3901 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcc
//
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