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""ë",#ä(  6B  ›GACNNNNNNGTC,CCAGGCCTGG,GATCCGGATC,CCWGGTACCWGG,CCANNNNNTGG,CCAGGNNNTGG,CCANNNCCTGG,CCAGGNCCTGG,GCACNNNAACGTTNNNGTGC,CACNNNGTG,CCNNNNNNNGG,AACNAACRYGTGCGTGC,GCACGCACRYGTTNGTT,GCACNAACRYGTTNGTGC,AACGCACRYGTGCGTT,GCACNNNNNNGTTSAACNNNNNNGTGC,GACNNNNNGTC,GAANNNNNNNTTGG,CCAANNNNNNNTTC,CACNNNNGTG,AACACNNNNGTGC,GCACNNNNGTGTT,GCANNNNNNTGC,GAACNNNNNNTCC,GGANNNNNNGTTC,CAGNNNCTG,CCAGGNNCTG,CAGNNCCTGG,CCWGGGCCC,GGGCCCWGG,CCWGGGCCCWGG,AACNNNNNNGTGCACNNNNNNGTT,GACNNNNNNTTYG,CRAANNNNNNGTC,GACNNNGTC,GAANNNNTTC,CCWGGTACC,GGTACCWGG,CCWGGWCCWGG,ACNNNNGTAYC,GRTACNNNNGT,CCWGGYRCCWGG,GAAGNNNNNNTAC,GTANNNNNNCTTC,AACGGCNNNGTGC,GCACNNNGCCGTT,GCACGGCNNNGTT,AACNNNGCCGTGC,CGANNNNNNTGC,GCANNNNNNTCG,CGATCNNNNTGC,GCANNNNGATCG,TGANNNNNNTCA,TGATCNNNNTCA,TGANNNNGATCA,TGATCNNGATCA,AACCTGCNNGTGC,GCACNNGCAGGTT,GCACCTGCNNGTT,AACNNGCAGGTGC,GCCNNNNNGGC,AACNNNNNNGTGCMC,GKGCACNNNNNNGTT,CCWGGNCCWGG,AACTGGGNNGTGC,GCACNNCCCAGTT,GCACTGGGNNGTT,AACNNCCCAGTGC,GACNNNNGTC,GAGNNNNNCTC,CCWGGTCTC,GAGACCWGG,GATNNNNATC,GATCNNNATC,GATNNNGATC,GATCNNGATC,CCWGGCGCCWGG,ACNNNNNCTCC,GGAGNNNNNGT,AACNNNNNNGTGCAG,CTGCACNNNNNNGTT,GATCATGATC,CCWGGNNCCWGG,AACTGGNNNGTGC,GCACNNNCCAGTT,GCACTGGNNNGTT,AACNNNCCAGTGC,AACNAACCGGTGCGTGC,GCACGCACCGGTTNGTT,GCACNAACCGGTTNGTGC,AACGCACCGGTGCGTT,GCANNNNNTGC,GCACNNNNNNGTTCGAACNNNNNNGTGC,CCTNNNNNAGG,GCNNNNNNNGC,CCANNNNNNTGG,CCAGGNNCCTGG,CCTGGCCAGG,CAANNNNNGTGG,CCACNNNNNTTG,TTTAAACNNNNNNGTGC,GCACNNNNNNGTTTAAA,GCACNNNNNNGTTTAAACNNNNNNGTGC,CCAGGNCCY,RGGNCCTGG,CCWGGGNCCTGG,CCAGGNCCCWGG,AAGNNNNNCTT,GAYNNNNNVTC,GABNNNNNRTC,GAYNNNNNRTC,GATCNNNNVTC,GABNNNNGATC,GAYNNNNGATC,GATCNNNNRTC,GATCNNNGATC,GTYAACNNNNNNGTGC,GCACNNNNNNGTTRAC,GCACNNNNNNGTTAACNNNNNNGTGC,GATCNGATC,TAACTATAACGGTCCTAAGGTAGCGA,TCGCTACCTTAGGACCGTTATAGTTA,CTCTCTTAAGGTAGC,GCTACCTTAAGAGAG,TAGGGATAACAGGGTAAT,ATTACCCTGTTATCCCTA,GTTAACNNNNNNGTGC,GCACNNNNNNGTTAAC,AACNAACGCGTGCGTGC,GCACGCACGCGTTNGTT,GCACNAACGCGTTNGTGC,AACGCACGCGTGCGTT,TCCAACNNNNNNGTGC,GCACNNNNNNGTTGGA,CAYNNNNRTG,AACAYNNNNGTGC,GCACNNNNRTGTT,GTTTAAACNNNNNNGTGC,GCACNNNNNNGTTTAAAC,CCWGGCGCC,GGCGCCWGG,CCWGGNNCC,GGNNCCWGG,GATCGCGATC,AACNAACATGTGCGTGC,GCACGCACATGTTNGTT,GCACNAACATGTTNGTGC,AACGCACATGTGCGTT,TGGCAAACAGCTATTATGGGTATTATGGGT,ACCCATAATACCCATAATAGCTGTTTGCCA,ATCTATGTCGGGTGCGGAGAAAGAGGTAATGAAATGG,CCATTTCATTACCTCTTTCTCCGCACCCGACATAGAT,AACNNGCACGTGCNNGTT,GAACNNNNNCTC,GAGNNNNNGTTC,CCAGGWCCY,RGGWCCTGG,CCWGGGWCCTGG,CCAGGWCCCWGG,CCAGGWCCTGG,TTATAACNNNNNNGTGC,GCACNNNNNNGTTATAA,GCACNNNNNNGTTATAACNNNNNNGTGC,GAACNNNNNNTAC,GTANNNNNNGTTC,GGCCNNNNNGGCC,AACNNACTAGTGC,GCACTAGTNNGTT,AACTAGTNNGTGC,GCACNNACTAGTT,AACNAACTAGTGCGTGC,GCACGCACTAGTTNGTT,GCACNAACTAGTTNGTGC,AACGCACTAGTGCGTT,AACNNNAACGTGCNNGTGC,GCACNNGCACGTTNNNGTT,TTAACNNNNNNGTGC,GCACNNNNNNGTTAA,CACNNNNNNTCC,GGANNNNNNGTG,CACNNNNGATCC,GGATCNNNNGTG,GATCTAGATC,CCANNNNNNNNNTGG,CCAGGCCT,AGGCCTGG,CCTCGAGG,GATCCGGA,TCCGGATC,GGCGCGCC,GCGATCGC,CCWGGWCC,GGWCCWGG,GATCGATC,GATCATGA,TCATGATC,GCGGCCGC,CCTGGCCR,YGGCCAGG,CCWGGNCC,GGNCCWGG,GGCCGGCC,RTGCGCAY,GATCNNGA,TCNNGATC,CGCGCGCG,CCTGGCCA,TGGCCAGG,CGCCGGCG,GTTTAAAC,GATCGCGA,TCGCGATC,TTAATTAA,VCTCGAGB,CCTGCAGG,CRCCGGYG,CGTCGACG,ATTTAAAT,GCCCGGGC,GATCTAGA,TCTAGATC,CACCTGC,GCAGGTG,CCTNAGG,GATCGAT,ATCGATC,CCTCAGC,GCTGAGG,GCTNAGC,CCTNAGC,GCTNAGG,GCTCTTC,GAAGAGC,GGTNACC,CGGWCCG,ACCWGGT,RGGNCCY,CCTGGAG,CTCCAGG,GGTGATC,GATCACC,GATCNGA,TCNGATC,GGGWCCC,GAAGATC,GATCTTC,CCCWGGG,TCCNGGA,TCCWGGA,RGGWCCY,TTATAA,AGGCCT,GACGTC,GTMKAC,TCCGGA,TGCGCA,ACCTGC,GCAGGT,GGTACC,AGTACT,GGYRCC,CCGCTC,GAGCGG,AACGTT,YGGCCR,RAATTY,CTGAAG,CTTCAG,CACGTG,GRCGYC,AGCGCT,CTTAAG,ACRYGT,ACCGGT,ACTAGT,CACGTC,GACGTG,GWGCWC,GTGCAC,CYCGRG,GGGCCC,ATTAAT,CCTAGG,TTCGAA,GCTAGC,GKGCMC,TGGCCA,GGATCC,GRGCYC,ATCGAT,CACGAG,CTCGTG,GGCGCC,GAAGAC,GTCTTC,GCATGC,GTATCC,GGATAC,TGATCA,ACTGGG,CCCAGT,CTRYAG,RGCGCY,AGATCT,GGNNCC,CTGGAG,CTCCAG,CTTGAG,CTCAAG,CGATCG,GGTCTC,GAGACC,YACGTR,CCNNGG,GAATGC,GCATTC,WCCGGW,GCAATG,CATTGC,RCCGGY,GCGCGC,GAGGAG,CTCCTC,CGGCCG,CCCAGC,GCTGGG,GTGCAG,CTGCAC,CGRYCG,CGTACG,CGTCTC,GAGACG,CCATGG,TCGCGA,GDGCHC,TGTACA,TCATGA,ACATGT,GTATAC,CCWWGG,CTCTTC,GAAGAG,GCNNGC,CCRYGG,GTTAAC,RCATGY,TACGTA,RGATCY,GCGATG,CATCGC,GCAGTG,CACTGC,CCCGGG,CCGCGG,TTTAAA,GGCGGA,TCCGCC,GAGCTC,GATATC,CTGRAG,CTYCAG,CAGCAG,CTGCTG,GAATTC,ATGCAT,CATATG,GTYRAC,AAGCTT,GTNNAC,TCNNGA,CAATTG,ACGCGT,TCCRAC,GTYGGA,GCCGGC,CMGCKG,GCCGAG,CTCGGC,CTCGAG,CTGCAG,CAGCTG,GTCGAC,CTYRAG,AATATT,GATCGA,TCGATC,GACCGA,TCGGTC,WGTACW,TARCCA,TGGYTA,TCTAGA,GGATC,GATCC,TTSAA,CCWGG,GTCTC,GAGAC,GCWGC,GGNCC,CCSGG,GGTGA,TCACC,GGWCC,GCAGC,GCTGC,CCATC,GATGG,ACGGC,GCCGT,GCNGC,CCNGG,GCATC,GATGC,ACTGG,CCAGT,GGATG,CATCC,CTCAG,CTGAG,GGGAC,GTCCC,ACNGT,CTNAG,CAGTG,CACTG,GACGC,GCGTC,CCCGC,GCGGG,GANTC,CGWCG,TCNGA,CCTTC,GAAGG,GCTCC,GGAGC,GTNAC,GAAGA,TCTTC,GAGTC,GACTC,GTSAC,GAWTC,GCSGC,CASTG,ATGAA,TTCAT,ACGGA,TCCGT,CGCG,CCGC,GCGG,GTAC,AGCT,GCGC,CTAG,GATC,GGCC,CCGG,CATG,RGCY,YATR,ACGT,TGCA,CCDG,CHGG,AATT,CCTC,GAGG,TTAA,ASST,TCGA                                                                                                                                                                                                                                                                                                                                                                "                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                            
                                                    
                                   	                                                                                                                           
                                           (                                                                                   0                                                                       
   	   &         
                        	         	      -      
         	               
               
   	            #      '                  !      	   	                                 %      4   ,      /   $          •   ‚      !   r   p   ?   (   #   @   �   '                                                                                                                                                                  "                                                                                                                                                                                                                                                                                
                  
             	                                           
               (                             0                         
 	 &   
        	   	  -  
   	     
     
 	    #  '      !  	 	           %  4 ,  / $    • ‚  ! r p ? ( # @ � '   <  ¡l  ¡Ž  ¡ô  §¦  ®F  ¸¬èUt#)ñ(I#q%_&a&p-	?	
ñ(ÃU+ö{j"°#m%3)×
{“&z#¶ÈÆ&%T&9¼%¸&Xw€|¢ÑpÍ

üc]bÕ¡Õ`
T"<°eù¢ùtQñz#Ã{’`ä 	
[´÷KwÚîÒZxziæÂïÄß6"T"t"v" "»"Ù"x&Þ&~'HI …4}ˆ×¼PÔ” �×Ô1%×Ô” Õ<Û£ìT&A6	P›¨{# jsñ$K'Ü  )
ÎÇT&  íÿÁÇT	¾â'/½Dó$ÚÉ%–ð @Îºx¯š ü 4'pCi@y²%%¯š/pmÉz´ä]" 8)ä]" @Î¯š ü 4'"ãÍÉ# $&l	vè�$‰&-ûwîN"½õï/å'ž!ùÖ|Ö‰xï—&þ	w
v\Lqž!"#�»Ò ¹"¬�ö$!ä$Sâ	*
xÐÇ¼Ö‘ê™!Ê!¹$} í ó
lÚö&²á"+).W4¢X5!$$¹&-'s(-¶¡ó EÙ4M¿þ	\6Gq' Hƒ“h»ýN!'Ú÷6<‘žêñÝB(¶(<¶(×	Æ«"C\Aœ W&„,!\³)Å&W<EW&ù&¶(Ç¼Öê©áêJE  #ú%W&?§»Ú þ	\qkc5ÕÂ&Œƒg­�˜Ùª«â!(‚ÆIæŒK�¿÷»³ã­!l",%ã'`þ	ô\ Hƒ“q¬"´"ÆoAc±„­	6	ì	þ	w
v\ÎLxq¯°š›ý ž!##"#ù%$&W&4'‚ø!ì%I'kÏê¾	s!ä@jh=:"”¯°N1%4þMI#D´Ö	×	v{Æoz{P¨"#Zñ´µÊ^ì!>%T& 'T' œi&´ó†-N~QëdÐ%¡‡ä	
›!â"ßÝÀ!ûRïõ	>*
x‘™!$ï	÷ÔÒ$

ô'²Óh*
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_'l	�$‰&ñ^ 'T')AO— %•%Ê%Á –Âð¶K�šŒ† ,"È&H ið 	E	ä	û	
 
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X!”$'B)UffrÈÓ´Æo?x'%ãDnâU#2KNÉ×	Æ"vç ˜L_R
½>z	«Ý&êç'-‚ø¶!¡ó EÙì%I'Hq ë¼PXÌÉX–$©%(�
è3qM%Õ%pÞa(%n<2l	�
:vè3e8§q:L3ÊÉ R!F#}#Ž#�$ç$M%Õ%‰&''¶( * ”�„Î!ƒåk½	ü
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This inhibition can be relieved by adding lactose or isopropyl-Î²-D-thiogalactopyranoside (IPTG).&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="15" name="CAP binding site" type="protein_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3167-3188" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;E. coli&lt;/i&gt; catabolite activator protein&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;CAP binding activates transcription in the presence of cAMP.&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="16" name="lac promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment name="-35" range="3203-3208" color="#ffffff" type="standard"/><Segment range="3209-3226" color="#ffffff" type="standard"/><Segment name="-10" range="3227-3233" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;promoter for the &lt;i&gt;E. coli lac&lt;/i&gt; operon&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="17" name="Cas9(N)" directionality="1" translationMW="66437.14" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="5816" maxFusedRunOn="5816" detectionMode="exactProteinMatch"><Segment range="3789-5507" 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;N-terminal portion of &lt;i&gt;Streptococcus pyogenes&lt;/i&gt;&amp;nbsp;Cas9 (&lt;a href=&quot;http://www.ncbi.nlm.nih.gov/pubmed/25643054&quot; title=&quot;http://www.ncbi.nlm.nih.gov/pubmed/25643054&quot;&gt;Zetsche et al., 2015&lt;/a&gt;)&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"/></Q></Feature><Feature recentID="18" name="AmpR" directionality="1" translationMW="31557.36" type="CDS" allowSegmentOverlaps="0" cleavageArrows="782" consecutiveTranslationNumbering="1" hitsStopCodon="1" detectionMode="exactProteinMatch"><Segment name="signal sequence" range="714-782" color="#ccffcc" type="standard" translated="1"/><Segment range="783-1574" 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="19" name="AmpR promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="609-713" 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></Features>
  *¨<?xml version="1.0"?><Primers nextValidID="15"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="Amp-R" sequence="ATAATACCGCGCCACATAGC" description="Ampicillin resistance gene, reverse primer"><BindingSite location="931-950" boundStrand="1" annealedBases="ATAATACCGCGCCACATAGC" meltingTemperature="58"><Component hybridizedRange="931-950" bases="ATAATACCGCGCCACATAGC"/></BindingSite><BindingSite simplified="1" location="931-950" boundStrand="1" annealedBases="ATAATACCGCGCCACATAGC" meltingTemperature="58"><Component hybridizedRange="931-950" bases="ATAATACCGCGCCACATAGC"/></BindingSite></Primer><Primer recentID="1" name="CYC1" sequence="GCGTGAATGTAAGCGTGAC" description="CYC1 transcription termination signal, reverse primer"><BindingSite location="8370-8388" boundStrand="1" annealedBases="GCGTGAATGTAAGCGTGAC" meltingTemperature="56"><Component hybridizedRange="8370-8388" bases="GCGTGAATGTAAGCGTGAC"/></BindingSite><BindingSite simplified="1" location="8370-8388" boundStrand="1" annealedBases="GCGTGAATGTAAGCGTGAC" meltingTemperature="56"><Component hybridizedRange="8370-8388" bases="GCGTGAATGTAAGCGTGAC"/></BindingSite></Primer><Primer recentID="2" name="F1ori-F" sequence="GTGGACTCTTGTTCCAAACTGG" description="F1 origin, forward primer"><BindingSite location="2085-2094" boundStrand="1" annealedBases="TCCAACTGG" meltingTemperature="13"><Component bases="GTGGACTCTTGT"/><Component hybridizedRange="2090-2094" bases="TCCAA"/><Component bases="A"/><Component hybridizedRange="2085-2088" bases="CTGG"/></BindingSite><BindingSite location="9102-9123" boundStrand="0" annealedBases="GTGGACTCTTGTTCCAAACTGG" meltingTemperature="57"><Component hybridizedRange="9102-9123" bases="GTGGACTCTTGTTCCAAACTGG"/></BindingSite><BindingSite simplified="1" location="2085-2094" boundStrand="1" annealedBases="TCCAACTGG" meltingTemperature="13"><Component bases="GTGGACTCTTGT"/><Component hybridizedRange="2090-2094" bases="TCCAA"/><Component bases="A"/><Component hybridizedRange="2085-2088" bases="CTGG"/></BindingSite><BindingSite simplified="1" location="9102-9123" boundStrand="0" annealedBases="GTGGACTCTTGTTCCAAACTGG" meltingTemperature="57"><Component hybridizedRange="9102-9123" bases="GTGGACTCTTGTTCCAAACTGG"/></BindingSite></Primer><Primer recentID="3" name="F1ori-R" sequence="AGGGAAGAAAGCGAAAGGAG" description="F1 origin, reverse primer"><BindingSite location="8892-8911" boundStrand="1" annealedBases="AGGGAAGAAAGCGAAAGGAG" meltingTemperature="57"><Component hybridizedRange="8892-8911" bases="AGGGAAGAAAGCGAAAGGAG"/></BindingSite><BindingSite simplified="1" location="8892-8911" boundStrand="1" annealedBases="AGGGAAGAAAGCGAAAGGAG" meltingTemperature="57"><Component hybridizedRange="8892-8911" bases="AGGGAAGAAAGCGAAAGGAG"/></BindingSite></Primer><Primer recentID="4" name="L4440" sequence="AGCGAGTCAGTGAGCGAG" description="L4440 vector, forward primer"><BindingSite location="3032-3049" boundStrand="0" annealedBases="AGCGAGTCAGTGAGCGAG" meltingTemperature="58"><Component hybridizedRange="3032-3049" bases="AGCGAGTCAGTGAGCGAG"/></BindingSite><BindingSite simplified="1" location="3032-3049" boundStrand="0" annealedBases="AGCGAGTCAGTGAGCGAG" meltingTemperature="58"><Component hybridizedRange="3032-3049" bases="AGCGAGTCAGTGAGCGAG"/></BindingSite></Primer><Primer recentID="5" name="M13 Forward" sequence="TGTAAAACGACGGCCAGT" description="In lacZ gene. Also called M13-F20 or M13 (-21) Forward"><BindingSite location="8648-8665" boundStrand="1" annealedBases="TGTAAAACGACGGCCAGT" meltingTemperature="57"><Component hybridizedRange="8648-8665" bases="TGTAAAACGACGGCCAGT"/></BindingSite><BindingSite simplified="1" location="8648-8665" boundStrand="1" annealedBases="TGTAAAACGACGGCCAGT" meltingTemperature="57"><Component hybridizedRange="8648-8665" bases="TGTAAAACGACGGCCAGT"/></BindingSite></Primer><Primer recentID="6" name="M13 Reverse" sequence="CAGGAAACAGCTATGAC" description="In lacZ gene. Also called M13-rev"><BindingSite location="3264-3280" boundStrand="0" annealedBases="CAGGAAACAGCTATGAC" meltingTemperature="49"><Component hybridizedRange="3264-3280" bases="CAGGAAACAGCTATGAC"/></BindingSite><BindingSite simplified="1" location="3264-3280" boundStrand="0" annealedBases="CAGGAAACAGCTATGAC" meltingTemperature="49"><Component hybridizedRange="3264-3280" bases="CAGGAAACAGCTATGAC"/></BindingSite></Primer><Primer recentID="7" name="M13/pUC Forward" sequence="CCCAGTCACGACGTTGTAAAACG" description="In lacZ gene"><BindingSite location="8657-8679" boundStrand="1" annealedBases="CCCAGTCACGACGTTGTAAAACG" meltingTemperature="61"><Component hybridizedRange="8657-8679" bases="CCCAGTCACGACGTTGTAAAACG"/></BindingSite><BindingSite simplified="1" location="8657-8679" boundStrand="1" annealedBases="CCCAGTCACGACGTTGTAAAACG" meltingTemperature="61"><Component hybridizedRange="8657-8679" bases="CCCAGTCACGACGTTGTAAAACG"/></BindingSite></Primer><Primer recentID="8" name="M13/pUC Reverse" sequence="AGCGGATAACAATTTCACACAGG" description="In lacZ gene"><BindingSite location="3245-3267" boundStrand="0" annealedBases="AGCGGATAACAATTTCACACAGG" meltingTemperature="58"><Component hybridizedRange="3245-3267" bases="AGCGGATAACAATTTCACACAGG"/></BindingSite><BindingSite simplified="1" location="3245-3267" boundStrand="0" annealedBases="AGCGGATAACAATTTCACACAGG" meltingTemperature="58"><Component hybridizedRange="3245-3267" bases="AGCGGATAACAATTTCACACAGG"/></BindingSite></Primer><Primer recentID="9" name="pBR322ori-F" sequence="GGGAAACGCCTGGTATCTTT" description="pBR322 origin, forward primer"><BindingSite location="92-105" boundStrand="0" annealedBases="CGCCTGTATCTTT" meltingTemperature="37"><Component bases="GGGAAA"/><Component hybridizedRange="92-96" bases="CGCCT"/><Component bases="G"/><Component hybridizedRange="98-105" bases="GTATCTTT"/></BindingSite><BindingSite location="2233-2252" boundStrand="0" annealedBases="GGGAAACGCCTGGTATCTTT" meltingTemperature="56"><Component hybridizedRange="2233-2252" bases="GGGAAACGCCTGGTATCTTT"/></BindingSite><BindingSite location="5087-5098" boundStrand="1" annealedBases="CCTGTATCTTT" meltingTemperature="26"><Component bases="GGGAAACG"/><Component hybridizedRange="5095-5098" bases="CCTG"/><Component bases="G"/><Component hybridizedRange="5087-5093" bases="TATCTTT"/></BindingSite><BindingSite simplified="1" location="92-105" boundStrand="0" annealedBases="CGCCTGTATCTTT" meltingTemperature="37"><Component bases="GGGAAA"/><Component hybridizedRange="92-96" bases="CGCCT"/><Component bases="G"/><Component hybridizedRange="98-105" bases="GTATCTTT"/></BindingSite><BindingSite simplified="1" location="2233-2252" boundStrand="0" annealedBases="GGGAAACGCCTGGTATCTTT" meltingTemperature="56"><Component hybridizedRange="2233-2252" bases="GGGAAACGCCTGGTATCTTT"/></BindingSite><BindingSite simplified="1" location="5087-5098" boundStrand="1" annealedBases="CCTGTATCTTT" meltingTemperature="26"><Component bases="GGGAAACG"/><Component hybridizedRange="5095-5098" bases="CCTG"/><Component bases="G"/><Component hybridizedRange="5087-5093" bases="TATCTTT"/></BindingSite></Primer><Primer recentID="10" name="pBRforEco" sequence="AATAGGCGTATCACGAGGC" description="pBR322 vectors, upsteam of EcoRI site, forward primer"><BindingSite location="9-27" boundStrand="1" annealedBases="AATAGGCGTATCACGAGGC" meltingTemperature="57"><Component hybridizedRange="9-27" bases="AATAGGCGTATCACGAGGC"/></BindingSite><BindingSite simplified="1" location="9-27" boundStrand="1" annealedBases="AATAGGCGTATCACGAGGC" meltingTemperature="57"><Component hybridizedRange="9-27" bases="AATAGGCGTATCACGAGGC"/></BindingSite></Primer><Primer recentID="11" name="pGEX 3'" sequence="CCGGGAGCTGCATGTGTCAGAGG" description="pGEX vectors, reverse primer"><BindingSite location="10553-10575" boundStrand="0" annealedBases="CCGGGAGCTGCATGTGTCAGAGG" meltingTemperature="66"><Component hybridizedRange="10553-10575" bases="CCGGGAGCTGCATGTGTCAGAGG"/></BindingSite><BindingSite simplified="1" location="10553-10575" boundStrand="0" annealedBases="CCGGGAGCTGCATGTGTCAGAGG" meltingTemperature="66"><Component hybridizedRange="10553-10575" bases="CCGGGAGCTGCATGTGTCAGAGG"/></BindingSite></Primer><Primer recentID="12" name="pRS-marker" sequence="CGGCATCAGAGCAGATTGTA" description="pRS vectors, use to sequence yeast selectable marker"><BindingSite location="10434-10453" boundStrand="1" annealedBases="CGGCATCAGAGCAGATTGTA" meltingTemperature="54"><Component hybridizedRange="10434-10453" bases="CGGCATCAGAGCAGATTGTA"/></BindingSite><BindingSite simplified="1" location="10434-10453" boundStrand="1" annealedBases="CGGCATCAGAGCAGATTGTA" meltingTemperature="54"><Component hybridizedRange="10434-10453" bases="CGGCATCAGAGCAGATTGTA"/></BindingSite></Primer><Primer recentID="13" name="T3" sequence="GCAATTAACCCTCACTAAAGG" description="T3 promoter, forward primer"><BindingSite location="3299-3319" boundStrand="0" annealedBases="GCAATTAACCCTCACTAAAGG" meltingTemperature="53"><Component hybridizedRange="3299-3319" bases="GCAATTAACCCTCACTAAAGG"/></BindingSite><BindingSite location="8620-8630" boundStrand="1" annealedBases="CTCACTAAGG" meltingTemperature="18"><Component bases="GCAATTAACC"/><Component hybridizedRange="8624-8630" bases="CTCACTA"/><Component bases="A"/><Component hybridizedRange="8620-8622" bases="AGG"/></BindingSite><BindingSite simplified="1" location="3299-3319" boundStrand="0" annealedBases="GCAATTAACCCTCACTAAAGG" meltingTemperature="53"><Component hybridizedRange="3299-3319" bases="GCAATTAACCCTCACTAAAGG"/></BindingSite><BindingSite simplified="1" location="8620-8630" boundStrand="1" annealedBases="CTCACTAAGG" meltingTemperature="18"><Component bases="GCAATTAACC"/><Component hybridizedRange="8624-8630" bases="CTCACTA"/><Component bases="A"/><Component hybridizedRange="8620-8622" bases="AGG"/></BindingSite></Primer><Primer recentID="14" name="T7" sequence="TAATACGACTCACTATAGGG" description="T7 promoter, forward primer"><BindingSite location="3309-3320" boundStrand="0" annealedBases="CTCACTAAGGG" meltingTemperature="28"><Component bases="TAATACGA"/><Component hybridizedRange="3309-3315" bases="CTCACTA"/><Component bases="T"/><Component hybridizedRange="3317-3320" bases="AGGG"/></BindingSite><BindingSite location="8619-8638" boundStrand="1" annealedBases="TAATACGACTCACTATAGGG" meltingTemperature="50"><Component hybridizedRange="8619-8638" bases="TAATACGACTCACTATAGGG"/></BindingSite><BindingSite simplified="1" location="3309-3320" boundStrand="0" annealedBases="CTCACTAAGGG" meltingTemperature="28"><Component bases="TAATACGA"/><Component hybridizedRange="3309-3315" bases="CTCACTA"/><Component bases="T"/><Component hybridizedRange="3317-3320" bases="AGGG"/></BindingSite><BindingSite simplified="1" location="8619-8638" boundStrand="1" annealedBases="TAATACGACTCACTATAGGG" meltingTemperature="50"><Component hybridizedRange="8619-8638" bases="TAATACGACTCACTATAGGG"/></BindingSite></Primer></Primers>
  ÷<Notes>
<UUID>a4bc6b53-7e0e-4682-8a5a-6f12c214bc54</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<CustomMapLabel>pEmpty_Control</CustomMapLabel>
<UseCustomMapLabel>1</UseCustomMapLabel>
<Description>Empty control for Cas9 experiments and to clone new guide RNAs for paper.</Description>
<Created UTC="4:0:0">2024.8.25</Created>
<LastModified UTC="6:16:38">2026.1.14</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
<References>
<Reference title="CRISPR-Cas9 Gene Editing in Yeast: A Molecular Biology and Bioinformatics Laboratory Module for Undergraduate and High School Students." doi="10.1128/jmbe.00106-21" type="Journal Article" journal="J Microbiol Biol Educ. 2021 May 31;22(2):e00106-21. doi: 10.1128/jmbe.00106-21. eCollection 2021 Fall." journalName="J Microbiol Biol Educ" issue="2" pubMedID="34594460" pages="e00106-21" authors="Sankaran SM, Smith JD, Roy KR" volume="22" date="2021-05-31- 31"/>
</References>
</Notes>
  Y     –         
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   )<?xml version="1.0"?><CustomEnzymeSets/>
