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pAC1453_pX-sgTelomere-25xPBSa

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pAC1453_pX-sgTelomere-25xPBSa載體質粒基本信息

質粒名稱: pAC1453_pX-sgTelomere-25xPBSa
出品公司: Addgene
目錄編號: 73695
存儲實驗室: Albert Cheng

載體骨架基本信息
載體名稱: pX330; pUC ori vector
載體類型: 哺乳細胞表達,CRISPR
載體抗性: 氨芐霉素100 μg/mL
生長溫度: 37 ℃
宿主菌: DH5a

插入基因信息
插入基因名稱: sgTelomere-25xPBSa
啟動子: U6
5’測序引物 gagggcctatttcccatgattcc
3’測序引物 gccatttgtctgcagaattggc

文獻引用方法
材料和方法部分: pAC1453_pX-sgTelomere-25xPBSa was a gift from Albert Cheng (Addgene plasmid # 73695 ; //n2t.net/addgene:73695 ; RRID:Addgene_73695)
參考文獻部分: Casilio: a versatile CRISPR-Cas9-Pumilio hybrid for gene regulation and genomic labeling. Cheng AW, Jillette N, Lee P, Plaskon D, Fujiwara Y, Wang W, Taghbalout A, Wang H. Cell Res. 2016 Feb;26(2):254-7. doi: 10.1038/cr.2016.3. Epub 2016 Jan 15. 10.1038/cr.2016.3 PubMed 26768771

pAC1453_pX-sgTelomere-25xPBSa質粒圖譜載體圖譜和pAC1453_pX-sgTelomere-25xPBSa載體序列質粒序列多克隆位點信息

pAC1453_pX-sgTelomere-25xPBSa載體圖譜

pAC1453_pX-sgTelomere-25xPBSa質粒載體簡介

pAC1453_pX-sgTelomere-25xPBSa質粒序列載體序列

LOCUS       pAC1453_pX-sgTelomere-25xPBSa               3498 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Vector expressing sgTelomere-25xPBSa.
ACCESSION   .
VERSION     .
KEYWORDS    pAC1453_pX-sgTelomere-25xPBSa
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3498)
  AUTHORS   Cheng AW, Jillette N, Lee P, Plaskon D, Fujiwara Y, Wang W, 
            Taghbalout A, Wang H
  TITLE     Casilio: a versatile CRISPR-Cas9-Pumilio hybrid for gene regulation 
            and genomic labeling.
  JOURNAL   Cell Res. 2016 Feb;26(2):254-7. doi: 10.1038/cr.2016.3. Epub 2016 
            Jan 15.
  PUBMED    26768771
REFERENCE   2  (bases 1 to 3498)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            //www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3498
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        9..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     9..29
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     180..199
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     repeat_region   761..901
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   761..890
                     /label=AAV2 ITR
     rep_origin      976..1431
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1063..1082)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1273..1294
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1448..1467)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1567..1589
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1627..1645)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1713..1817
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1818..2678
                     /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(2036..2055)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2849..3437
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3338..3357
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggcgcgccga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
       61 ttagagagat aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt
      121 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
      181 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt
      241 ggaaaggacg aaacaccgtt agggttaggg ttagggttag tttaagagct atgctggaaa
      301 cagcatagca agtttaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
      361 ggtgccaatt gggtctccag attgtatgta gcctgtatgt agcctgtatg tagcctgtat
      421 gtagcctgta tgtaagattg tatgtagcct gtatgtagcc tgtatgtagc ctgtatgtag
      481 cctgtatgta agattgtatg tagcctgtat gtagcctgta tgtagcctgt atgtagcctg
      541 tatgtaagat tgtatgtagc ctgtatgtag cctgtatgta gcctgtatgt agcctgtatg
      601 taagattgta tgtagcctgt atgtagcctg tatgtagcct gtatgtagcc tgtatgtaag
      661 atcttttttt gttttagagc tagaaatagc aagttaaaat aaggctagtc cgtagcgcgt
      721 gcgccaattc tgcagacaaa tggcggcgcg ccgcggccgc aggaacccct agtgatggag
      781 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
      841 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
      901 gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt
      961 caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     1021 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     1081 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     1141 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     1201 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     1261 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct
     1321 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     1381 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca
     1441 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     1501 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     1561 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     1621 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     1681 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1741 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     1801 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     1861 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     1921 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     1981 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2041 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     2101 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2161 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2221 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2281 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2341 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2401 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2461 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2521 ccggtgagcg tggaagccgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2581 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     2641 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     2701 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     2761 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     2821 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     2881 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     2941 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3001 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3061 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3121 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3181 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3241 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3301 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3361 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3421 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3481 ggccttttgc tcacatgt
//

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