| 质粒名称: | pHelper-T7IS1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 140631 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pUC19 |
| 完整质粒大小: | 9066bp |
| 载体类型: | Bacterial Expression, CRISPR ; Transposon |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | ShTnsB, ShTnsC, ShTniQ, ShCas12k |
| 物种来源: | Scytonema hofmanni |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | Unknown (unknown if destroyed) |
| 3’克隆位点: | Unknown (unknown if destroyed) |
|
文献引用方法 |
|
| 材料和方法部分: | pHelper-T7IS1 was a gift from Sheng Yang (Addgene plasmid # 140631 ; http://n2t.net/addgene:140631 ; RRID:Addgene_140631) |
| 参考文献部分: | Multicopy chromosomal integration using CRISPR-associated transposases. Zhang Y, Sun X, Wang Q, Xu J, Dong F, Yang S, Yang J, Zhang Z, Qian Y, Chen J, Zhang J, Liu Y, Tao R, Jiang Y, Yang J, Yang S. ACS Synth Biol. 2020 Jun 17. doi: 10.1021/acssynbio.0c00073. 10.1021/acssynbio.0c00073 PubMed 32551502 |
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 pHelper-T7IS1 9066 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Expresses ShCAST under the T7 promoter and an empty sgRNA.
ACCESSION .
VERSION .
KEYWORDS pHelper-T7IS1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9066)
AUTHORS Zhang Y, Sun X, Wang Q, Xu J, Dong F, Yang S, Yang J, Zhang Z, Qian
Y, Chen J, Zhang J, Liu Y, Tao R, Jiang Y, Yang J, Yang S
TITLE Multicopy chromosomal integration using CRISPR-associated
transposases.
JOURNAL ACS Synth Biol. 2020 Jun 17. doi: 10.1021/acssynbio.0c00073.
PUBMED 32551502
REFERENCE 2 (bases 1 to 9066)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..9066
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 86..105
/label=T7
/note="T7 promoter, forward primer"
promoter 86..104
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 105..129
/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)."
terminator 5584..5655
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
promoter 5670..5703
/label=J23119 promoter
/note="bacterial promoter (Registry of Standard Biological
Parts BBa_J23119)"
terminator 5958..5985
/label=T7Te terminator
/note="phage T7 early transcription terminator"
promoter 6012..6116
/gene="bla"
/label=AmpR promoter
CDS 6117..6977
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(6335..6354)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7148..7736
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7637..7656
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7946..9028)
/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 8990..9009
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(9029..40)
/gene="lacI"
/label=lacI promoter
ORIGIN
1 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
61 ttatgcgact cctgcattag gaaattaata cgactcacta taggggaatt gtgagcggat
121 aacaattccc ctgtagaaat aattttgttt aactttaata aggagatata ccatgaacag
181 tcagcaaaat cctgatttag ctgttcatcc cttggcaatt cctatggaag gcttactagg
241 agaaagtgct acaactcttg agaagaatgt aattgccaca caactctcag aggaagccca
301 agtaaagcta gaggtaatcc aaagtttact ggaaccctgc gatcgcacaa cttatgggca
361 aaagttgcgg gaagcagcag agaaactaaa tgtatcgttg cgaacggtac aaaggttggt
421 gaaaaactgg gaacaagatg gcttagtcgg actcactcaa acaagtaggg ctgataaagg
481 aaaacaccgc attggtgagt tttgggaaaa cttcattacc aaaacctaca aggagggtaa
541 caagggaagt aaacgtatga cccctaaaca agttgctctc agagtcgagg ctaaagcccg
601 tgaattaaaa gactctaagc cgcccaatta caaaaccgtg ttacgggtat tagcacccat
661 tttggaaaag caacaaaaag ccaagagtat ccgcagtcct ggttggagag gaactacgct
721 ttcggttaaa acccgtgaag gaaaagattt atcggttgat tacagtaacc atgtttggca
781 atgtgaccat acccgcgtgg atgtgttgct ggtagatcaa catggtgaaa ttttaagtcg
841 tccctggcta acaacagtaa ttgatactta ctctcgttgc attatgggta tcaacttggg
901 ctttgatgca cccagttctg gggtagtagc attagcgtta cgccatgcaa ttctaccaaa
961 gcgttacggt tccgagtaca aactgcattg tgagtgggga acctatggaa aaccagaaca
1021 tttttatact gatggcggta aagactttcg ctctaaccac ttgagtcaga ttggggcgca
1081 attgggattt gtctgtcatt tacgcgatcg cccttctgaa ggtggagtag tagaacgtcc
1141 cttcaaaaca ttaaatgacc aactattttc aacgcttcct gggtacaccg gatctaatgt
1201 gcaggaacgc ccagaagatg cagagaagga cgcaagactt actttgcgag aactagaaca
1261 gttacttgtg cgttacatcg tagatcgtta caaccaaagt attgatgcgc ggatgggcga
1321 ccaaacgcgc tttgagcgtt gggaagcagg attgcctaca gtgccagtac caataccaga
1381 acgagatttg gatatttgtt taatgaagca gtcacggcgc actgtgcaaa gaggtggttg
1441 tttgcagttt cagaatttaa tgtatcgggg ggaatatttg gcaggttatg ccggagaaac
1501 tgtcaactta aggtttgacc ccagagacat tacaacaatt ttggtttatc gccaggaaaa
1561 caatcaggaa gtatttctga ctcgcgctca cgctcaaggt ttggagacag agcaactggc
1621 attagatgag gctgaggcag caagtcgcag actccgtacc gcagggaaaa ctatcagtaa
1681 ccaatcatta ttgcaagaag ttgttgaccg cgatgctctt gtcgctacca agaaaagccg
1741 taaggagcgt caaaaattgg aacagactgt tttgcgatct gctgctgttg atgaaagtaa
1801 tagagaatcc ttgccttctc aaatagttga accagatgaa gtggaatcta cagaaacggt
1861 tcactctcaa tacgaagaca ttgaggtgtg ggactatgaa caacttcgtg aagaatatgg
1921 gttttaaaca atgacagaag ctcaggcgat cgccaagcag ttgggtgggg taaaaccgga
1981 tgatgagtgg ttacaagctg aaattgctcg tctcaagggt aagagcattg tgcctttaca
2041 gcaggtaaaa actctccatg attggttaga tggcaagcgc aaggcaagaa aatcttgccg
2101 agtagttggg gaatcgagaa ctggcaagac agttgcttgt gatgcctaca gatacaggca
2161 caaacctcag caggaagctg gacgacctcc aactgtgcct gtcgtttata ttcgacctca
2221 ccaaaaatgt ggccccaagg atttgtttaa aaagattact gagtacctca agtatcgggt
2281 aacaaaaggg actgtatctg attttcgaga taggacgata gaagtactca agggttgtgg
2341 cgtagagatg ctaattattg atgaagctga ccgtctcaag cctgaaactt ttgctgatgt
2401 gcgagatatt gccgaagatt taggaattgc tgtggtactg gtaggaacag accgtttgga
2461 tgcggtaatt aagcgggatg agcaggttct cgaacgcttt cgggcgcatc ttcgctttgg
2521 taaattgtcg ggagaggatt ttaagaacac cgtagaaatg tgggaacaaa tggttttgaa
2581 actgccagta tcttctaatc taaagagcaa ggagatgcta cggattctca cgtcagcaac
2641 tgaaggctac attggtcgcc ttgatgagat tcttagggaa gctgcaattc gttccttatc
2701 aagaggattg aagaagattg acaaggctgt tttacaggaa gtagctaagg agtacaaatg
2761 atagaagcac cagatgttaa accttggcta ttcttgatta aaccctatga aggggaaagc
2821 ctgagccact ttcttggcag gttcagacgt gccaaccatt tatccgcaag tggattgggt
2881 actttggcag gaattggtgc tatagtggca cgttgggaaa gatttcattt taatcctcgc
2941 cctagtcagc aagaattgga agcgatcgca tctgtagtag aagtggatgc tcaaaggtta
3001 gcccagatgt taccgcctgc tggagtggga atgcagcatg agccaattcg cttgtgtggg
3061 gcttgttatg ccgagtcgcc ttgtcaccga attgaatggc agtacaagtc ggtgtggaag
3121 tgcgatcgcc atcaactcaa gattttagca aagtgtccaa actgtcaagc accttttaaa
3181 atgcctgcgc tgtgggagga tgggtgctgt cacagatgta ggatgccgtt tgcagaaatg
3241 gcaaagctac agaaggtttg atgataaaac cagaaaaagg tgtgaaatta actaagtccc
3301 tgaattgatc tggttgtcca aaaaatttgt gcgatcgcat ggcaagatta ttcctactat
3361 tgatgtggcg tgagtcggat aacttgctct aatgggatcc cggttaatac caatagatca
3421 gatatgagta ataaacatta cgacctttac ctgtggtatt cactgttggc acgacaacac
3481 ctttatcgcg ccaatactgt agctgacgac gcgaacaatt agtgatttga gccgcttctg
3541 tgcttgtgta gaattttcct tccatagaaa tcatcctaca gaagtagttt tttaaattta
3601 cattttttaa aagcacatta tgtattcatg tgatattatt tttttatgag tcaaataact
3661 attcaagctc gacttatttc ctttgaatca aaccgccaac aactctggaa gttgatggca
3721 gatttaaaca cgccgttaat taacgaactg ctttgccagt taggtcaaca ccccgacttc
3781 gagaagtggc aacaaaaggg taaactcccg tctaccgttg tgagccagtt atgtcaacct
3841 ctcaaaactg accctcgctt tgcaggtcag cccagccgtt tatatatgtc ggcaattcat
3901 attgtggact acatctacaa gtcctggctg gctatacaga aacggcttca acagcagcta
3961 gatggaaaga cgcgctggct agaaatgctc aatagcgatg ctgaattagt agaacttagt
4021 ggtgacactt tagaggctat tcgtgtcaaa gctgctgaaa ttttggcaat agctatgcca
4081 gcatctgagt cagatagcgc ttcacctaaa gggaaaaaag gtaaaaagga gaaaaaaccc
4141 tcatcttcta gccctaagcg tagtttatcc aagacattat ttgacgctta ccaagaaacg
4201 gaagatatca agagccgtag cgccatcagc tacctgttaa aaaatggctg caaacttact
4261 gacaaagaag aagattcaga aaaatttgct aaacgtcgtc gtcaagttga aatccaaatt
4321 caaaggctta ccgaaaagtt aataagtcgg atgcctaaag gtcgagattt gaccaatgct
4381 aaatggttgg agacactctt gactgctaca accactgttg ctgaagacaa cgcccaagcc
4441 aaacgctggc aggatattct gttaactcga tcaagttctc tcccattccc ccttgttttt
4501 gaaaccaacg aggatatggt ttggtcaaag aatcaaaagg gtaggctgtg tgttcacttc
4561 aatggcttaa gcgatttaat ttttgaggtg tactgcggca atcgtcaact tcactggttt
4621 caacgcttcc tagaagacca acagactaaa cgcaaaagca aaaatcagca ttctagcggc
4681 ttgttcacac tcagaaatgg tcatctagtt tggcttgaag gtgagggtaa aggggaacct
4741 tggaatcttc accacttgac cctttactgc tgtgttgaca atcgcttgtg gacagaggag
4801 ggaacagaaa tcgttcgcca agagaaagca gatgaaatta ctaaattcat cacaaacatg
4861 aagaagaaaa gcgatctaag cgatacacag caagctttga ttcaacgtaa acaatcaaca
4921 cttactcgaa taaacaattc ctttgagcgt cctagccaac ccctttatca aggtcaatca
4981 cacattttgg ttggagtaag cctgggacta gaaaaacctg ccacagtagc agtagtagat
5041 gcgatcgcca acaaagtctt ggcttaccgg agtattaaac aattacttgg cgacaattac
5101 gaactgctaa atcgccagag acgacaacag cagtacctat ctcacgaacg ccacaaagca
5161 caaaaaaact tctctcccaa tcaatttgga gcatctgagt tagggcaaca tatagacaga
5221 ttattagcta aagcaattgt agcgttagcg agaacctaca aagctggcag tattgtcttg
5281 cccaagttag gggatatgcg ggaggttgtc caaagtgaaa ttcaagctat agcagaacaa
5341 aaatttcccg gttatattga aggtcagcaa aaatatgcca aacagtaccg ggttaatgtt
5401 catcggtgga gctacggcag attaattcaa agcattcaaa gtaaagcagc tcaaacagga
5461 attgtgattg aggagggaaa acaacctatt cgaggtagtc cccacgacaa agcaaaggaa
5521 ttagcacttt ctgcttacaa tctccgccta actaggcgaa gttaagtaag gatctccagg
5581 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
5641 tcggtgaacg ctctctacta gagagaccct gacagctagc tcagtcctag gtataatgct
5701 agcatattaa tagcgccgca attcatgctg cttgcagcct ctgaattttg ttaaatgagg
5761 gttagtttga ctgtataaat acagtcttgc tttctgaccc tggtagctgc tcaccctgat
5821 gctgctgtca atagacagga taggtgcgct cccagcaata agggcgcgga tgtactgctg
5881 tagtggctac tgaatcaccc ccgatcaagg gggaacccta aatgggttga aagttttatg
5941 ttcagataat gcccgatggc tcaccttcgg gtgggccttt ctgcgcaggt ggcacttttc
6001 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
6061 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
6121 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
6181 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
6241 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
6301 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
6361 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
6421 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
6481 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
6541 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
6601 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
6661 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
6721 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
6781 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
6841 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
6901 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
6961 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
7021 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
7081 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
7141 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
7201 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
7261 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
7321 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
7381 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
7441 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
7501 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
7561 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
7621 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
7681 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
7741 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct caacgccaag
7801 ctccaaaacg atctcaagaa gatcatctta ttaattcatg ccccgcgccc accggaagga
7861 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
7921 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
7981 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
8041 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
8101 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
8161 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
8221 ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
8281 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
8341 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
8401 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
8461 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
8521 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
8581 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
8641 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
8701 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
8761 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
8821 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
8881 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
8941 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
9001 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
9061 gctatc
//
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