LOCUS pET-dCas9-VP64-6 9544 bp DNA circular UNA 21-JAN-2015
DEFINITION .
ACCESSION urn.local...1n-47xzcrz
VERSION urn.local...1n-47xzcrz
KEYWORDS .
SOURCE
ORGANISM
.
FEATURES Location/Qualifiers
misc_feature 1..70
/Original_Bases
/label
/note="Geneious type: Editing History Insertion"
misc_feature 2..29
/Imported_from="Pla
sMapper"
/Transferred_From="lacO reg"
/Transferred_Similarity="100.00%"
/label="lacO reg"
/note="Geneious type: regulatory sequence"
misc_feature 30..35
/Recognition_pattern="T^CTAGA"
/annotation_group="TCTAGA"
/label="XbaI"
/note="Geneious type: restriction site"
gene 47..63
/Imported_from="Pla
sMapper"
/Transferred_From="T7_transl_en_RBS"
/Transferred_Similarity="100.00%"
/label="T7_transl_en_RBS"
gene 71..4174
/created_by="User"
/modified_by="User"
/label="dCas9 gene"
misc_feature 71..73
/created_by="User"
/label="Start ATG"
misc_feature 98..100
/created_by="User"
/modified_by="User"
/label="D10A mutation"
/note="Geneious type: sequence variant"
misc_feature 2588..2590
/created_by="User"
/label="H840A mutation"
/note="Geneious type: sequence variant"
misc_feature 4181..4201
/created_by="User"
/label="SV40 NLS"
sig_peptide 4208..4279
/created_by="User"
/label="3X FLAG tag"
repeat_unit 4295..4453
/created_by="User"
/label="VP64"
misc_feature 4454..9544
/Original_Bases
/label
/note="Geneious type: Editing History Insertion"
primer_bind 4454..4471
/Color="#ff9911"
/created_by="User"
/label="6xHis"
primer_bind 4472..4474
/created_by="User"
/Color="#ff3333"
/modified_by="User"
/label="STOP"
primer_bind 4475..4477
/created_by="User"
/Color="#ff3333"
/modified_by="User"
/label="STOP"
misc_feature 4478..4483
/Recognition_pattern="C^TCGAG"
/annotation_group="CTCGAG"
/label="XhoI"
/note="Geneious type: restriction site"
misc_feature 4795..4800
/Recognition_pattern="GCC^GGC"
/annotation_group="GCCGGC"
/label="NaeI"
/note="Geneious type: restriction site"
misc_feature 5177..5182
/Recognition_pattern="TTT^AAA"
/annotation_group="TTTAAA"
/label="DraI"
/note="Geneious type: restriction site"
misc_feature 5196..5201
/Recognition_pattern="TTT^AAA"
/annotation_group="TTTAAA"
/label="DraI"
/note="Geneious type: restriction site"
misc_feature 5427..5437
/Recognition_pattern="GCCNNNN^NGGC"
/annotation_group="GCCNNNNNGGC"
/label="BglI"
/note="Geneious type: restriction site"
misc_feature 5680..5685
/Recognition_pattern="CGAT^CG"
/annotation_group="CGATCG"
/label="PvuI"
/note="Geneious type: restriction site"
misc_feature 5791..5796
/Recognition_pattern="AGT^ACT"
/annotation_group="AGTACT"
/label="ScaI"
/note="Geneious type: restriction site"
misc_feature 5888..5893
/Recognition_pattern="TTT^AAA"
/annotation_group="TTTAAA"
/label="DraI"
/note="Geneious type: restriction site"
misc_feature 5980..5985
/Recognition_pattern="G^TGCAC"
/annotation_group="GTGCAC"
/label="ApaLI"
/note="Geneious type: restriction site"
promoter complement(6143..6171)
/Imported_from="Pla
sMapper"
/Transferred_From="amp prom"
/Transferred_Similarity="100.00%"
/label="amp prom"
misc_feature 6604..6609
/Recognition_pattern="G^TGCAC"
/annotation_group="GTGCAC"
/label="ApaLI"
/note="Geneious type: restriction site"
misc_feature 7104..7109
/Recognition_pattern="G^TGCAC"
/annotation_group="GTGCAC"
/label="ApaLI"
/note="Geneious type: restriction site"
gene complement(7292..7483)
/Imported_from="Pla
sMapper"
/Transferred_From="ROP"
/Transferred_Similarity="100.00%"
/label="ROP"
misc_feature 7329..7334
/Recognition_pattern="CAG^CTG"
/annotation_group="CAGCTG"
/label="PvuII"
/note="Geneious type: restriction site"
misc_feature 8091..8096
/Recognition_pattern="CAG^CTG"
/annotation_group="CAGCTG"
/label="PvuII"
/note="Geneious type: restriction site"
misc_feature 8184..8189
/Recognition_pattern="CAG^CTG"
/annotation_group="CAGCTG"
/label="PvuII"
/note="Geneious type: restriction site"
misc_feature 8278..8283
/Recognition_pattern="GTT^AAC"
/annotation_group="GTTAAC"
/label="HpaI"
/note="Geneious type: restriction site"
misc_feature 8600..8606
/Recognition_pattern="G^GTNACC"
/annotation_group="GGTNACC"
/label="BstEII"
/note="Geneious type: restriction site"
misc_feature 8802..8807
/Recognition_pattern="G^TGCAC"
/annotation_group="GTGCAC"
/label="ApaLI"
/note="Geneious type: restriction site"
misc_feature 9311..9316
/Recognition_pattern="GCATG^C"
/annotation_group="GCATGC"
/label="SphI"
/note="Geneious type: restriction site"
misc_feature 9472..9477
/Recognition_pattern="GCC^GGC"
/annotation_group="GCCGGC"
/label="NaeI"
/note="Geneious type: restriction site"
misc_feature 9506..9511
/Recognition_pattern="AT^CGAT"
/annotation_group="ATCGAT"
/label="ClaI"
/note="Geneious type: restriction site"
promoter 9528..2
/Imported_from="Pla
sMapper"
/Transferred_From="T7 prom"
/Transferred_Similarity="100.00%"
/label="T7 prom"
ORIGIN
1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
61 gagatatacc atggataaga aatactcaat aggcttagct atcggcacaa atagcgtcgg
121 atgggcggtg atcactgatg aatataaggt tccgtctaaa aagttcaagg ttctgggaaa
181 tacagaccgc cacagtatca aaaaaaatct tataggggct cttttatttg acagtggaga
241 gacagcggaa gcgactcgtc tcaaacggac agctcgtaga aggtatacac gtcggaagaa
301 tcgtatttgt tatctacagg agattttttc aaatgagatg gcgaaagtag atgatagttt
361 ctttcatcga cttgaagagt cttttttggt ggaagaagac aagaagcatg aacgtcatcc
421 tatttttgga aatatagtag atgaagttgc ttatcatgag aaatatccaa ctatctatca
481 tctgcgaaaa aaattggtag attctactga taaagcggat ttgcgcttaa tctatttggc
541 cttagcgcat atgattaagt ttcgtggtca ttttttgatt gagggagatt taaatcctga
601 taatagtgat gtggacaaac tatttatcca gttggtacaa acctacaatc aattatttga
661 agaaaaccct attaacgcaa gtggagtaga tgctaaagcg attctttctg cacgattgag
721 taaatcaaga cgattagaaa atctcattgc tcagctcccc ggtgagaaga aaaatggctt
781 atttgggaat ctcattgctt tgtcattggg tttgacccct aattttaaat caaattttga
841 tttggcagaa gatgctaaat tacagctttc aaaagatact tacgatgatg atttagataa
901 tttattggcg caaattggag atcaatatgc tgatttgttt ttggcagcta agaatttatc
961 agatgctatt ttactttcag atatcctaag agtaaatact gaaataacta aggctcccct
1021 atcagcttca atgattaaac gctacgatga acatcatcaa gacttgactc ttttaaaagc
1081 tttagttcga caacaacttc cagaaaagta taaagaaatc ttttttgatc aatcaaaaaa
1141 cggatatgca ggttatattg atgggggagc tagccaagaa gaattttata aatttatcaa
1201 accaatttta gaaaaaatgg atggtactga ggaattattg gtgaaactaa atcgtgaaga
1261 tttgctgcgc aagcaacgga cctttgacaa cggctctatt ccccatcaaa ttcacttggg
1321 tgagctgcat gctattttga gaagacaaga agacttttat ccatttttaa aagacaatcg
1381 tgagaagatt gaaaaaatct tgacttttcg aattccttat tatgttggtc cattggcgcg
1441 tggcaatagt cgttttgcat ggatgactcg gaagtctgaa gaaacaatta ccccatggaa
1501 ttttgaagaa gttgtcgata aaggtgcttc agctcaatca tttattgaac gcatgacaaa
1561 ctttgataaa aatcttccaa atgaaaaagt actaccaaaa catagtttgc tttatgagta
1621 ttttacggtt tataacgaat tgacaaaggt caaatatgtt actgaaggaa tgcgaaaacc
1681 agcatttctt tcaggtgaac agaagaaagc cattgttgat ttactcttca aaacaaatcg
1741 aaaagtaacc gttaagcaat taaaagaaga ttatttcaaa aaaatagaat gttttgatag
1801 tgttgaaatt tcaggagttg aagatagatt taatgcttca ttaggtacct accatgattt
1861 gctaaaaatt attaaagata aagatttttt ggataatgaa gaaaatgaag atatcttaga
1921 ggatattgtt ttaacattga ccttatttga agatagggag atgattgagg aaagacttaa
1981 aacatatgct cacctctttg atgataaggt gatgaaacag cttaaacgtc gccgttatac
2041 tggttgggga cgtttgtctc gaaaattgat taatggtatt agggataagc aatctggcaa
2101 aacaatatta gattttttga aatcagatgg ttttgccaat cgcaatttta tgcagctgat
2161 ccatgatgat agtttgacat ttaaagaaga cattcaaaaa gcacaagtgt ctggacaagg
2221 cgatagttta catgaacata ttgcaaattt agctggtagc cctgctatta aaaaaggtat
2281 tttacagact gtaaaagttg ttgatgaatt ggtcaaagta atggggcggc ataagccaga
2341 aaatatcgtt attgaaatgg cacgtgaaaa tcagacaact caaaagggcc agaaaaattc
2401 gcgagagcgt atgaaacgaa tcgaagaagg tatcaaagaa ttaggaagtc agattcttaa
2461 agagcatcct gttgaaaata ctcaattgca aaatgaaaag ctctatctct attatctcca
2521 aaatggaaga gacatgtatg tggaccaaga attagatatt aatcgtttaa gtgattatga
2581 tgtcgatgcc attgttccac aaagtttcct taaagacgat tcaatagaca ataaggtctt
2641 aacgcgttct gataaaaatc gtggtaaatc ggataacgtt ccaagtgaag aagtagtcaa
2701 aaagatgaaa aactattgga gacaacttct aaacgccaag ttaatcactc aacgtaagtt
2761 tgataattta acgaaagctg aacgtggagg tttgagtgaa cttgataaag ctggttttat
2821 caaacgccaa ttggttgaaa ctcgccaaat cactaagcat gtggcacaaa ttttggatag
2881 tcgcatgaat actaaatacg atgaaaatga taaacttatt cgagaggtta aagtgattac
2941 cttaaaatct aaattagttt ctgacttccg aaaagatttc caattctata aagtacgtga
3001 gattaacaat taccatcatg cccatgatgc gtatctaaat gccgtcgttg gaactgcttt
3061 gattaagaaa tatccaaaac ttgaatcgga gtttgtctat ggtgattata aagtttatga
3121 tgttcgtaaa atgattgcta agtctgagca agaaataggc aaagcaaccg caaaatattt
3181 cttttactct aatatcatga acttcttcaa aacagaaatt acacttgcaa atggagagat
3241 tcgcaaacgc cctctaatcg aaactaatgg ggaaactgga gaaattgtct gggataaagg
3301 gcgagatttt gccacagtgc gcaaagtatt gtccatgccc caagtcaata ttgtcaagaa
3361 aacagaagta cagacaggcg gattctccaa ggagtcaatt ttaccaaaaa gaaattcgga
3421 caagcttatt gctcgtaaaa aagactggga tccaaaaaaa tatggtggtt ttgatagtcc
3481 aacggtagct tattcagtcc tagtggttgc taaggtggaa aaagggaaat cgaagaagtt
3541 aaaatccgtt aaagagttac tagggatcac aattatggaa agaagttcct ttgaaaaaaa
3601 tccgattgac tttttagaag ctaaaggata taaggaagtt aaaaaagact taatcattaa
3661 actacctaaa tatagtcttt ttgagttaga aaacggtcgt aaacggatgc tggctagtgc
3721 cggagaatta caaaaaggaa atgagctggc tctgccaagc aaatatgtga attttttata
3781 tttagctagt cattatgaaa agttgaaggg tagtccagaa gataacgaac aaaaacaatt
3841 gtttgtggag cagcataagc attatttaga tgagattatt gagcaaatca gtgaattttc
3901 taagcgtgtt attttagcag atgccaattt agataaagtt cttagtgcat ataacaaaca
3961 tagagacaaa ccaatacgtg aacaagcaga aaatattatt catttattta cgttgacgaa
4021 tcttggagct cccgctgctt ttaaatattt tgatacaaca attgatcgta aacgatatac
4081 gtctacaaaa gaagttttag atgccactct tatccatcaa tccatcactg gtctttatga
4141 aacacgcatt gatttgagtc agctaggagg tgacggttct cccaagaaga agaggaaagt
4201 ctcgagcgac tacaaagacc atgacggtga ttataaagat catgacatcg attacaagga
4261 tgacgatgac aaggctgcag gaggcggtgg aagcgggcgc gccgacgcgc tggacgattt
4321 cgatctcgac atgctgggtt ctgatgccct cgatgacttt gacctggata tgttgggaag
4381 cgacgcattg gatgactttg atctggacat gctcggctcc gatgctctgg acgatttcga
4441 tctcgatatg ttacatcacc accaccatca ctaatgactc gagtctggta aagaaaccgc
4501 tgctgcgaaa tttgaacgcc agcacatgga ctcgtctact agcgcagctt aattaaccta
4561 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
4621 gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt
4681 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
4741 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
4801 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
4861 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
4921 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
4981 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
5041 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
5101 aacaaaatat taacgtttac aatttctggc ggcacgatgg catgagatta tcaaaaagga
5161 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
5221 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
5281 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
5341 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
5401 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
5461 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
5521 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
5581 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
5641 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
5701 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
5761 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
5821 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
5881 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
5941 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
6001 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
6061 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatcat
6121 gattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
6181 aaaaataaac aaataggtca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
6241 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
6301 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
6361 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
6421 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
6481 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
6541 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
6601 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
6661 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
6721 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
6781 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
6841 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
6901 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
6961 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
7021 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
7081 cggtatttca caccgcatat atggtgcact ctcagtacaa tctgctctga tgccgcatag
7141 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
7201 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
7261 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
7321 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
7381 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
7441 taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag
7501 ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac
7561 gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg
7621 tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata
7681 cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa
7741 tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca
7801 ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc
7861 gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca
7921 acgacaggag cacgatcatg ctagtcatgc cccgcgccca ccggaaggag ctgactgggt
7981 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
8041 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
8101 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt
8161 cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt
8221 tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt
8281 aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc
8341 gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg
8401 ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga
8461 aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga
8521 gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc
8581 gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta
8641 ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat
8701 aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga
8761 tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag
8821 gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg
8881 cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca
8941 acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa
9001 ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc
9061 tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat
9121 aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc
9181 ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg
9241 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
9301 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
9361 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
9421 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
9481 gatgcgtccg gcgtagagga tcgagatcga tctcgatccc gcgaaattaa tacgactcac
9541 tata
//