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                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                     	                                                                                        
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   #   	               	      	                                    2       	   -            ~   H            !         r                                                                                                                                                                                                                                                                                                                                                                                                                                  	                                
 
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                
        
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 # 	     	  	            2   	 -    ~ H    !   r   yä  zÔ  zî  {B  ~  „n  Šœh `QžŽÅ< ÞÙ™š‹aóŽÅMû³ÊrcÖ
�A	ð†`±·³»ãlã
²~SXø=~#Úúæï¾¾ceBXÀ ÙóÀ ò ù  Êu<�e’n
M<>\¥äÈ#åe¨VÚ”
7Ì‚1€±c­€f±úc€±c�BiM	`ÿfžé7èbBŽ{7�
ë7jøE	º�
Þû);àvpž9¾9©	B£ ì7JßÕ¾©	uMÎ@; 6û
Öq6û
Ö9¾9©	ìŒØþÙÓE@ŽkDDÝ*j
d	þ.jáÐå?åFø Jo_¢.ë4µ¦¯I¾‘oÈƒ�’
ëCJì·%’�™Ú	•€	ú¸ø 2Jt…_€þ(8sž	ñ
R	ë#	¾ Á|
Ñ%®¾ %Èý	
™¹@0¦¾ 	%oƒ�’
ÍÁÑÚ3
Ÿ˜ ø J_„
P´·ò¼³€#.	“	9y®	•¬ßv¦žb	L
ürô³Yø J(8s›_mßLõø Í¾¿©	ª	JC£oí_¢ .ë	04’»ŽÜšÃ½�Q[ºUpÔ!~×mTª«M	oÈÉû�
Êa˜™k@ALñ$æ‹¤½Y@Û
mŽ¹L‹žT¸|}Ìi®ÀÈCO¾…	b
ùH	 !ÈC~”<×lG@@ vñ$I¾ Á	|
ÑÚ%®\»«R	ëûÊñ$/Í¿	É	p£q}lz• )=AÃx|¨m•«¯¹Ï<¨¥®d7¤úOÖñ$2 R	ëhÍ¥áLõlwÍÙÙêw
MØ�JñUÈ™þàó§®«3ÅÁÑÔ€	ú¸šÃ½àQ�ïck°ƒT÷÷Ä ÙèˆÓë41ZÝKf¾ ÙK��&èƒVˆÓE�%~ÓŒPì@I¯Îþ­E	
€
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-P\»òúv›ŽºxŽ'ÔZ¼q»¾«2F©a,	Ã} ?:“$>9?4á³ãŸßv¦b	L
ü?:“9ÙÓ�ž	ñ
QºU[pÔ¿ŠÑm «“Î‡	•	–1´ jáŽÐj	åã?©åF ÁãAÓÍñáÉ
lÍÙÎ¦	I	"xÚªëo7�
ü
×a¡îAæŒ í"nHl¨A@ zTª«v6 Tßu<¨,’•eö;ŒžA‚…S›	l+:¿:ª	 
‚C¤¬í¼!’áòÐ8< _ ÈÉ™šaMf}r•# ¥¸
«ÎJB3OX(‡ÉÓ¶$:¿:ª	‚íòÐ6 ß¨’e‚…S	lTu<,•ö;ŒžA›
é
<æŒ'³$j„�ºÑÊ±\	r®1%E6 £ Tßu<¨.,’•˜5ùe‡]7ö;Œž¡²A‚…S›î	hMl< _ oÈÉ7�
ü
×a™š¡îAaMæŒf}r•¦ ò eäC_[ Ý K'’1‘	“	9�~‹?z”Œ @	¯	À# –7îZ N¨
&‘�Î	9
=³(‚ÚT	(ïKê<iÕwUþ—=¸ ù¨¼þMYOÁ©	üÀ\V	å	‹
î! C “�EÚ	G
ˆ
cÞBÅâVy¸ vÆØùÔï¨²Kc¼þê<iÕwUáMþ—=Y•,‘¡K™.—ò’Uƒ}Šæ]ÅQ“y 3çu†�¯° ‰ )T
 ×|(¦«‘,Òûý¿
3~‹¾ÖQyF‡Ò^c �Žè%	hÄPjq , © ‹Žð) §ºLh»ß-		ì
t·ùxºV
L[©
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  l<?xml version="1.0"?><Features nextValidID="11"><Feature recentID="0" name="Cas9" directionality="2" translationMW="158444.81" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="699" maxFusedRunOn="699" detectionMode="exactProteinMatch"><Segment range="750-4853" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="locus_tag"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;generates RNA-guided double strand breaks in DNA&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;Cas9 (Csn1) endonuclease from the&amp;nbsp;&lt;i&gt;Streptococcus pyogenes&lt;/i&gt;&amp;nbsp;Type II CRISPR/Cas system&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD"/></Q></Feature><Feature recentID="1" name="AmpR" directionality="2" translationMW="31558.16" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="6967" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1" detectionMode="exactProteinMatch"><Segment range="6176-6967" color="#ccffcc" type="standard" translated="1"/><Segment name="signal sequence" range="6968-7036" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to ampicillin, carbenicillin, and related antibiotics&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;Î²-lactamase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"/></Q></Feature><Feature recentID="2" name="ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5417-6005" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;high-copy-number ColE1/pMB1/pBR322/pUC origin of replication&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="3" name="Xenopus globin 3'-UTR" directionality="2" type="3'UTR" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="541-666" color="#a6acb3" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;3'-UTR of the major beta-globin gene of &lt;i&gt;Xenopus laevis&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="4" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7037-7141" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="5" name="nucleoplasmin NLS" directionality="2" translationMW="1739.18" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="699" maxFusedRunOn="699" detectionMode="exactProteinMatch"><Segment range="702-749" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;bipartite nuclear localization signal from nucleoplasmin&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="KRPAATKKAGQAKKKK"/></Q></Feature><Feature recentID="6" name="Xenopus globin 5'-UTR" directionality="2" type="5'UTR" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4923-4965" color="#a6acb3" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;translational enhancer from the 5'-UTR of the major beta-globin gene of &lt;i&gt;Xenopus laevis&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="SV40 NLS" directionality="2" translationMW="883.15" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" consecutiveNumberingStartsFrom="126" maxRunOn="699" maxFusedRunOn="699" detectionMode="exactProteinMatch"><Segment range="4878-4898" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="locus_tag"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;nuclear localization signal of SV40 (simian virus 40) large T antigen&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="PKKKRKV"/></Q></Feature><Feature recentID="8" name="SP6 promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="417-435" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage SP6 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="T7 promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4978-4996" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for bacteriophage T7 RNA polymerase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="10" name="Kozak sequence" type="regulatory" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4901-4910" color="#a6acb3" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;vertebrate consensus sequence for strong initiation of translation (&lt;a href=&quot;https://www.ncbi.nlm.nih.gov/pubmed/3313277&quot; title=&quot;https://www.ncbi.nlm.nih.gov/pubmed/3313277&quot;&gt;Kozak, 1987&lt;/a&gt;)&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="regulatory_class"><V predef="other"/></Q></Feature></Features>
   —<?xml version="1.0"?><Primers nextValidID="0"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/></Primers>
  ”<Notes>
<UUID>206037f6-ad64-4dc5-9fed-e4813825df06</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<CustomMapLabel>pXT7-Cas9</CustomMapLabel>
<UseCustomMapLabel>1</UseCustomMapLabel>
<Created UTC="9:28:1">2018.1.16</Created>
<LastModified UTC="4:28:15">2025.8.26</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  Y     K         
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