| 质粒名称: | pACYCT2 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 45799 |
| 存储实验室: | Luc Ponchon |
|
载体骨架基本信息 |
|
| 载体名称: | pACYCT2 |
| 空载体大小: | 3975 bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | 氯霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | XL1 Blue |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 克隆方法: | 限制性内切酶 |
|
文献引用方法 |
|
| 材料和方法部分: | pACYCT2 was a gift from Luc Ponchon (Addgene plasmid # 45799 ; http://n2t.net/addgene:45799 ; RRID:Addgene_45799) |
| 参考文献部分: | Co-expression of RNA-protein complexes in Escherichia coli and applications to RNA biology. Ponchon L, Catala M, Seijo B, El Khouri M, Dardel F, Nonin-Lecomte S, Tisne C. Nucleic Acids Res. 2013 Aug;41(15):e150. doi: 10.1093/nar/gkt576. Epub 2013 Jun 26. 10.1093/nar/gkt576 PubMed 23804766 |
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载体基本信息
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载体序列
LOCUS pACYCT2 3975 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pACYCT2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3975)
AUTHORS Ponchon L, Catala M, Seijo B, El Khouri M, Dardel F, Nonin-Lecomte
S, Tisne C
TITLE Co-expression of RNA-protein complexes in Escherichia coli and
applications to RNA biology.
JOURNAL Nucleic Acids Res. 2013 Aug;41(15):e150. doi: 10.1093/nar/gkt576.
Epub 2013 Jun 26.
PUBMED 23804766
REFERENCE 2 (bases 1 to 3975)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..3975
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 3..27
/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)."
CDS 83..100
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
promoter 197..225
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 233..249
/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)."
CDS 334..378
/codon_start=1
/product="affinity and epitope tag derived from pancreatic
ribonuclease A"
/label=S-Tag
/translation="KETAAAKFERQHMDS"
primer_bind complement(416..434)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 430..477
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(697..1356)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 1271..1290
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(1357..1459)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
primer_bind complement(1851..1868)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1985..2529)
/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(2065..2084)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2754..3836)
/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"
primer_bind 3798..3817
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(3837..3914)
/gene="lacI"
/label=lacI promoter
ORIGIN
1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag
121 ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc gaacagaaag
181 taatcgtatt gtacagttga caattaatca tcggctcgta taatgtgtgg aattgtgagc
241 ggataacaat tttaaggaga tatacatatg gcagatctca attggatatc ggccggccac
301 gcgatcgctg acgtcggtac cctcgagtct ggtaaagaaa ccgctgctgc gaaatttgaa
361 cgccagcaca tggactcgtc tactagcgca gcttaattaa cctaggctgc tgccaccgct
421 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
481 aaacctcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta
541 aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtcg
601 aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag gcgtagcacc
661 aggcgtttaa gggcaccaat aactgcctta aaaaaattac gccccgccct gccactcatc
721 gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac agacggcatg
781 atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat
841 agtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa
901 actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata
961 ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa
1021 atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt
1081 gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg ccatacggaa
1141 ctccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg
1201 cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata
1261 ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat
1321 atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa
1381 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga
1441 acctcttacg tgccgatcaa cgtctcattt tcgccaaaag ttggcccagg gcttcccggt
1501 atcaacaggg acaccaggat ttatttattc tgcgaagtga tcttccgtca caggtattta
1561 ttcggcgcaa agtgcgtcgg gtgatgctgc caacttactg atttagtgta tgatggtgtt
1621 tttgaggtgc tccagtggct tctgtttcta tcagctgtcc ctcctgttca gctactgacg
1681 gggtggtgcg taacggcaaa agcaccgccg gacatcagcg ctagcggagt gtatactggc
1741 ttactatgtt ggcactgatg agggtgtcag tgaagtgctt catgtggcag gagaaaaaag
1801 gctgcaccgg tgcgtcagca gaatatgtga tacaggatat attccgcttc ctcgctcact
1861 gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga acggggcgga
1921 gatttcctgg aagatgccag gaagatactt aacagggaag tgagagggcc gcggcaaagc
1981 cgtttttcca taggctccgc ccccctgaca agcatcacga aatctgacgc tcaaatcagt
2041 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tcccctggcg gctccctcgt
2101 gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt
2161 gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta
2221 tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt
2281 ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag
2341 gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac
2401 tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa
2461 aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg cagaccaaaa
2521 cgatctcaag aagatcatct tattaatcag ataaaatatt tctagatttc agtgcaattt
2581 atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgta attctcatgt
2641 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg
2701 gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
2761 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
2821 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
2881 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
2941 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
3001 ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg cagcccggac
3061 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
3121 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
3181 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
3241 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
3301 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
3361 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
3421 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
3481 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
3541 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
3601 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
3661 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
3721 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
3781 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
3841 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
3901 cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaaatt
3961 aatacgactc actat
//
