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T_7]¾Ç±,ª¬à ¹�¦ôµÏÖ'—�`	ë
5Û«ê ÞÞ24 «B�á1-b_ù®³uA¥Brw�	Èn0 è ý }®&;’»3AHgt:X”®Óâ �«Øh}ŒÝ	Ÿ
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Ž'²ðºE\w–.ÎOÈÓ ƒ’y,ðCÈ5	¯Öhn5[Z‰A‹à@�¨ú¸6˜Ïs M©5–·À>QiŸ%nUÎÞR6ì<`$ ²U‚´<¨«SÇùÝWéíÛ
zÝ£<”ô(Qh*±mx{3IL  a Ð f&53Z¹
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9‘ÁNà¥·m@¹jÎe¾V»ÍØ$ ^ §ÍEž ö¤)[W�2	]	Â
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~ë�Ó—Ï³•-Ž -µ ßæaŒAøF$¹ê�»ÄqÞ ‚d?QV:?¾ö*	7¿ ‡?4¼1q | ´<§Âv™l	�	³	×	û	B
eYb¾—ß#8M%É� ~ É 7y¢!žUQgƒ`£XksÏ©g� ¨ ÈQ¢ÉZ�<wz+Ùú&ÌDÄÊú … 5hãVû(Ç{“
ûÕJ…Dý& Su,/ÐSb›2eÄ@¶¹Ç!%.:DO}G=j¬ÆCFÚ^‘! Vìƒ,�ÅaC	˜j’›ÉÚã>“Õåƒ(�—BO²A® ­(i°
	óRÀw%U}œ¸å j/Fe1õ)HT7l-L´! Ÿ ªä~,óÎ&–œS6\ªé­°¶t@¤Aqv3Žñm* |­L~;Gf§x¨ÆÒÿ€•ªVg|ì‹ž
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gm4Z~Š'g)°;N_Ç Ï £¨o	“	¶	Ú	þ	!
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gvü.ú3±¨$Ä× $ ] ”�î Dö¶ÃZ^©Ðß(V€„£U	¶kÄ ÞèIidºÌ× �Lÿ3å>Kud}Xa€†²€º,p „ § tÇâax¡d?zú;µ-fù„ËÙ]Cù! ž ëã}òh%Ä›¯
[è‘šÙâ(=GSk$£'@Kpu|Ž±# “ÿe¦Â§ÅÑþ©¨ÏtŠT	n	’	µ	Ù	ý	 
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fjû-ù32‰§¯  ¦ þ`|üý#$sxyde9:ø�™Øá1R?Jotyz{B  -î  qGACNNNNNNGTC,CCAGGCCTGG,GATCCGGATC,CCWGGTACCWGG,CCANNNNNTGG,CCAGGNNNTGG,CCANNNCCTGG,CCAGGNCCTGG,GCACNNNAACGTTNNNGTGC,CACNNNGTG,CCNNNNNNNGG,AACNAACRYGTGCGTGC,GCACGCACRYGTTNGTT,GCACNAACRYGTTNGTGC,AACGCACRYGTGCGTT,GACNNNNNGTC,GAANNNNNNNTTGG,CCAANNNNNNNTTC,CACNNNNGTG,AACACNNNNGTGC,GCACNNNNGTGTT,GCANNNNNNTGC,GAACNNNNNNTCC,GGANNNNNNGTTC,CAGNNNCTG,CCAGGNNCTG,CAGNNCCTGG,CCWGGGCCC,GGGCCCWGG,CCWGGGCCCWGG,AACNNNNNNGTGCACNNNNNNGTT,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,GCACCCANNNGTT,AACNNNTGGGTGC,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,CACCCA,TCGGTC,TGGGTG,WGTACW,TARCCA,TGGYTA,TCTAGA,GGATC,GATCC,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,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,AATT,CCTC,GAGG,TTAA,ASST,TCGA                                         #                                                                                                                                                         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                                                 "                                                                                                                           3                
                                                                     #                   	                                                          	                     	                           	   1   !   
      	                                          
               	                        
                        &   &                   1         Š   A               "      W                      #                                                                                                                                                  ,                                                                                                                                                                                                                                                                                                    	                   	           
                 "                                               3      
                         #       	                    	       	         	 1 ! 
  	              
     	        
        & &       1   Š A     "  W   nk  o“  o»  oñ  tY  xÉ  ~É¼xu�ê„ … Ï �È`a›§ä§ýž
	�ÇOJjƒ£¤'J|}œýºÌ¢êSµ«CÖ¡U‰w'œ|yü%Ò%Ò·€¬ôªÏ §ž
” +wô%K
nw€'œÀá´½P�	¸
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for the &lt;i&gt;tetR&lt;/i&gt; and &lt;i&gt;tetA&lt;/i&gt; genes&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="6" name="tet operator" type="protein_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2414-2432" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;tetracycline repressor TetR&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;tetO&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;bacterial operator O&lt;sub&gt;2&lt;/sub&gt; for the &lt;i&gt;tetR&lt;/i&gt; and &lt;i&gt;tetA&lt;/i&gt; genes&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="tight TRE promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2406-2720" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;Tet-responsive promoter&amp;nbsp;P&lt;sub&gt;Tight&lt;/sub&gt;, consisting of seven tet operator&amp;nbsp;sequences followed by the minimal CMV promoter&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="miR-30a loop" directionality="1" type="ncRNA" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2894-2908" color="#8fbc8f" type="standard"/><Q name="ncRNA_class"><V predef="miRNA"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;loop from the&amp;nbsp;the 71-nt precursor of the human miR-30a microRNA (&lt;a href=&quot;http://www.ncbi.nlm.nih.gov/pubmed/12086629&quot; 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(or &lt;i&gt;nptII&lt;/i&gt;)&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to neomycin, kanamycin, and G418 (GeneticinÂ®)&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase from Tn5&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"/></Q></Feature><Feature recentID="14" name="ori" directionality="1" type="rep_origin" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5726-6314" color="#ffff00" type="standard"/><Q name="direction"><V text="RIGHT"/></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="15" name="f1 ori" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3344-3799" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;f1 bacteriophage origin of replication; 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   —<?xml version="1.0"?><Primers nextValidID="0"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/></Primers>
  ±<Notes>
<UUID>80944109-d4e9-44ac-858b-99499766b9d5</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<CustomMapLabel>pEF1a-tTA-Advanced-TRE-miR-30a loop-Neo</CustomMapLabel>
<UseCustomMapLabel>1</UseCustomMapLabel>
<Created UTC="16:0:0">2017.5.21</Created>
<LastModified UTC="1:3:50">2025.9.20</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  Y      K         
?????                                                                                                       ÿ d      DO  d              E              ÿÿÿÿYô? P                                                                                                                                                              ©<StrandColors><TopStrand><ColorRange range="0..6374" colors="black"/></TopStrand><BottomStrand><ColorRange range="0..6374" colors="black"/></BottomStrand></StrandColors>   3<?xml version="1.0"?><EnzymeVisibilities vals=""/>
   )<?xml version="1.0"?><CustomEnzymeSets/>
