• 规格：20μl质粒
• 启动子：U6
• 原核抗性：氨苄青霉素Amp
• 克隆菌株：大肠杆菌Stbl3
• 质粒简介：           AAV-U6gRNA1-U6gRNA2-TnT-Cre是一个哺乳细胞腺相关病毒Cre和2×gRNA表达质粒。
• 质粒图谱1： png/1-1G226103939336.png

质粒序列： 点击查看序列

LOCUS       Exported                5958 bp ds-DNA     circular SYN 26-DEC-2017

DEFINITION  synthetic circular DNA

FEATURES             Location/Qualifiers

     source          1..5958

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     promoter        1..241

                     /label=U6 promoter

                     /note="RNA polymerase III promoter for human U6 snRNA"

     misc_RNA        278..353

                     /label=gRNA scaffold

                     /note="guide RNA scaffold for the Streptococcus pyogenes 

                     CRISPR/Cas9 system"

     promoter        366..606

                     /label=U6 promoter

                     /note="RNA polymerase III promoter for human U6 snRNA"

     misc_RNA        633..708

                     /label=gRNA scaffold

                     /note="guide RNA scaffold for the Streptococcus pyogenes 

                     CRISPR/Cas9 system"

     misc_feature    721..1134

                     /label=gallus TnT promoter

     intron          1260..1392

                     /label=chimeric intron

                     /note="chimera between introns from human beta-globin and 

                     immunoglobulin heavy chain genes"

     promoter        1437..1455

                     /label=T7 promoter

                     /note="promoter for bacteriophage T7 RNA polymerase"

     CDS             1467..1487

                     /codon_start=1

                     /product="nuclear localization signal of SV40 (simian virus

                     40) large T antigen"

                     /label=SV40 NLS

                     /translation="PKKKRKV"

     misc_feature    1488..2516

                     /label=Cre

     polyA_signal    2554..3030

                     /label=hGH poly(A) signal

                     /note="human growth hormone polyadenylation signal"

     repeat_region   3070..3210

                     /label=AAV2 ITR

                     /note="inverted terminal repeat of adeno-associated virus 

                     serotype 2"

     rep_origin      3285..3740

                     /direction=RIGHT

                     /label=f1 ori

                     /note="f1 bacteriophage origin of replication; arrow 

                     indicates direction of (+) strand synthesis"

     promoter        4022..4126

                     /gene="bla"

                     /label=AmpR promoter

     CDS             4127..4987

                     /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"

     rep_origin      5158..5746

                     /direction=RIGHT

                     /label=ori

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 

                     replication"

ORIGIN

        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag

       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga

      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat

      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga

      241 cgaaacaccg actgcaggtg gaattccacc tgctcaggtt ttagagctag aaatagcaag

      301 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgctttttta

      361 cgcgtgaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta

      421 gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac

      481 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact

      541 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga

      601 aaggacgaaa caccggaaga gcgagctctt ctgttttaga gctagaaata gcaagttaaa

      661 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttctagt

      721 gggataaaag cagtctgggc tttcacatga cagcatctgg ggctgcggca gagggtcggg

      781 tccgaagcgc tgccttatca gcgtccccag ccctgggagg tgacagctgg ctggcttgtg

      841 tcagcccctc gggcactcac gtatctccgt ccgacgggtt taaaatagca aaactctgag

      901 gccacacaat agcttgggct tatatgggct cctgtggggg aagggggagc acggaggggg

      961 ccggggccgc tgctgccaaa atagcagctc acaagtgttg cattcctctc tgggcgccgg

     1021 gcacattcct gctggctctg cccgccccgg ggtgggcgcc ggggggacct taaagcctct

     1081 gccccccaag gagcccttcc cagacagccg ccggcaccca ccgctccgtg ggacgatccc

     1141 cgaagctcta gagctttatt gcggtagttt atcacagtta aattgctaac gcagtcagtg

     1201 cttctgacac aacagtctcg aacttaagct gcagaagttg gtcgtgaggc actgggcagg

     1261 taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac

     1321 agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct

     1381 ttctctccac aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat

     1441 acgactcact ataggctagc atggtgccca agaagaagag gaaagtctcc aacctgctga

     1501 ctgtgcacca aaacctgcct gccctccctg tggatgccac ctctgatgaa gtcaggaaga

     1561 acctgatgga catgttcagg gacaggcagg ccttctctga acacacctgg aagatgctcc

     1621 tgtctgtgtg cagatcctgg gctgcctggt gcaagctgaa caacaggaaa tggttccctg

     1681 ctgaacctga ggatgtgagg gactacctcc tgtacctgca agccagaggc ctggctgtga

     1741 agaccatcca acagcacctg ggccagctca acatgctgca caggagatct ggcctgcctc

     1801 gcccttctga ctccaatgct gtgtccctgg tgatgaggag aatcagaaag gagaatgtgg

     1861 atgctgggga gagagccaag caggccctgg cctttgaacg cactgacttt gaccaagtca

     1921 gatccctgat ggagaactct gacagatgcc aggacatcag gaacctggcc ttcctgggca

     1981 ttgcctacaa caccctgctg cgcattgccg aaattgccag aatcagagtg aaggacatct

     2041 cccgcaccga tggtgggaga atgctgatcc acattggcag gaccaagacc ctggtgtcca

     2101 cagctggtgt ggagaaggcc ctgtccctgg gggttaccaa gctggtggag agatggatct

     2161 ctgtgtctgg tgtggctgat gaccccaaca actacctgtt ctgccgggtc agaaagaatg

     2221 gtgtggctgc cccttctgcc acctcccaac tgtccacccg ggccctggaa gggatctttg

     2281 aggccaccca ccgcctgatc tatggtgcca aggatgactc tgggcagaga tacctggcct

     2341 ggtctggcca ctctgccaga gtgggtgctg ccagggacat ggccagggct ggtgtgtcca

     2401 tccctgaaat catgcaggct ggtggctgga ccaatgtgaa cattgtgatg aactacatca

     2461 gaaacctgga ctctgagact ggggccatgg tgaggctgct cgaggatggg gactgaaagc

     2521 ttatcgatac cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca

     2581 gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa

     2641 ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg

     2701 gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg

     2761 gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt

     2821 caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg

     2881 ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc

     2941 tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag

     3001 gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg

     3061 agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc

     3121 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag

     3181 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc

     3241 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg

     3301 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc

     3361 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc

     3421 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct

     3481 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac

     3541 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac

     3601 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat

     3661 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa

     3721 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata

     3781 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct

     3841 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt

     3901 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata

     3961 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt

     4021 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag

     4081 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca

     4141 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc

     4201 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat

     4261 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc

     4321 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg

     4381 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc

     4441 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat

     4501 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga

     4561 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc

     4621 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc

     4681 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt

     4741 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc

     4801 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc

     4861 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca

     4921 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca

     4981 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt

     5041 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta

&nbs