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tgcggcattttgccttcctgtttttgctcacccagaaacgctggtgaaagtaaaagatgctgaagatcagttgggtgcacgagtgggttacatcgaactggatctcaacagcggtaagatccttgagagttttcgccccgaagaacgttttccaatgatgagcacttttaaagttctgctatgtggcgcggtattatcccgtattgacgccgggcaagagcaactcggtcgccgcatacactattctcagaatgacttggttgagtactcaccagtcacagaaaagcatcttacggatggcatgacagtaagagaattatgcagtgctgccataaccatgagtgataacactgcggccaacttacttctgacaacgatcggaggaccgaaggagctaaccgcttttttgcacaacatgggggatcatgtaactcgccttgatcgttgggaaccggagctgaatgaagccataccaaacgacgagcgtgacaccacgatgcctgtagcaatggcaacaacgttgcgcaaactattaactggcgaactacttactctagcttcccggcaacaattaatagactggatggaggcggataaagttgcaggaccacttctgcgctcggcccttccggctggctggtttattgctgataaatctggagccggtgagcgtgggtctcgcggtatcattgcagcactggggccagatggtaagccctcccgtatcgtagttatctacacgacggggagtcaggcaactatggatgaacgaaatagacagatcgctgagataggtgcctcactgattaagcattggtaactgtcagaccaagtttactcatatatactttagattgatttaaaacttcatttttaatttaaaaggatctaggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgagttttcgttccactgagcgtcagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcgcagataccaaatactgttcttctagtgtagccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgtgagctatgagaaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggccttttgctcacatgt  3  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,GDGCHC,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,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,GCCGGC,CAATTG,ACGCGT,TCCRAC,GTYGGA,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,RGCR,YGCY,YATR,ACGT,TGCA,CCDG,CHGG,AATT,CCTC,GAGG,TTAA,ASST,TCGA                                        (                                                                                                                                                                                                                                                                                                                          #                                                       
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                                                                                                                                                                                                                                                                           	                   
                 
 
  (                                           .                                                                       	  
 < '             
    	 
      
    
   
  "        & *  ! /   9    s  O   e a  $ !  b   7  9@  9j  9  >.  C  M	fx	( p   ;yz4PU	f	x		nIE-uF5 hUW 	6	4#(
|
X1Kaz# Em		;#  `mvyZ8Sqbb!5 )hU5  )6hU5 )hU)6)(WSY~e EH0,g  	& zk	<hk	<HO
Jo^GKaWq	wyw 7Q		f	Q=tg  }%\"	
;'

-	3	K	+ Q	ww=a- _4E"
E' y36Z	E{NK
m KE ^g %
;gKP"

`%sQ		Z
S(9 	d
2Q	DL{   7kf8JKoQQ			'/+ gKEP"
 ^ 
YyMhKb$' z<"Z>Y	}	.NJ%{2 42qi4z W fKm		a< Ww	  QG}}	r_"}	N@    O S W  >rv?mf
VZKOSAOzn<}	.	_ZU<FQ ' {< n"

 -	3	K	
Yi:
\	 
OdEW	& x,27{g	

DLcz- ?	\ <
~ 	 ayK-Mbh3kvPebrw		$<F0R 0^?&P8

+-7N [?MgEK%Vd2?WH} 
  	S-R#k9<} 9KLp_}	kH,' ( A b{|)LAXFO^L	fo|   	-#}
 SRk9<p ( 32[Ep'	T			 
 #    ;B|0H$			S!	-RqV#k9<}' ( A [?MgEK%Vdb{|)LAX2?'-_{Sh&
7(	:[3@jc	(.G lR	=ie \'	uUWS`
Eh G lR	=i\Ie'I  X;\4A %_
#Z'%-w=, S	+mP	J>m?Cs )I#	k2R)[`- Fr		vMTnSj		
\f.c l,	J	|e|A
 lR
S&f.O5+K}PV/&2BU/n:

@kN		`II	[)`K}PV/2BU )#k2R#   ;B|0H$		!	qV I  %X_
#;\Z'%4A8	=lxE64fT-W X  I={/2g	rL*xz*a$  o<%	f		"6Q 
1W J>Bm|0?CHs	q+f#cs`	x	   7w1		#

l>Q7-  W Y  H^[z3;1fhR		qKM+EMy)z`{"v  <n.0=kB	'|?GR" D h  X:Lq~`&]~ 	#	r		
kH <`-( B  5b|{*LA3Xr`z#8J~				
[_V	 " D h     k7Xh:Lq~`oO&Q]~#lw 	#	1	7	r	z			

#

0kRHg@gl <`}R>Q7A-98- + 2 N R V   +G ADZgly2>Brx0GJpe	Q	w!&/Ye/DL,s(DJRhn%a*Ez!l   % ?  9p_pw4u,<<i&	=	G	S	g			K


	$S	UYb>T_ :NRVry 7@FNR[pyn%2*^z ?U!ajm %;>=\}`z#8Jr|7Y~				
0



-F^][_bVF#ZDgJ%{xt u   !F		t
]T"q  ( B   5Xs	[?x				
T
Mg,:EK[e%8Vdb|B{*L!A3c/,3@Xr2?P ( B  5Ti'u\f@N		


NuhFL&We%b|{8*LA3 )_Xr3@ .(Y


*wpF rkQ	9	Y	t	
>5@"<BD<a  `  #|Vj*Nou{ Aim	p			a^_( b.		O

y 	 6     C%7vC]Vw		1			#



{fxl	nt>Q` 7u-`Tgm?&



p   -<AlignableSequences trimStringency="Medium"/>  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  )<?xml version="1.0"?><Features nextValidID="19"><Feature recentID="0" name="FRT3" directionality="2" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2601-2634" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;FLP recombinase from the &lt;i&gt;Saccharomyces cerevisiae&lt;/i&gt; 2μ plasmid&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;supports FLP-mediated excision with other FRT3 sites (&lt;a href=&quot;https://pubmed.ncbi.nlm.nih.gov/25605786/&quot;&gt;Bosch et al., 2015&lt;/a&gt;)&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="1" name="FRT3" directionality="1" type="protein_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2032-2065" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;FLP recombinase from the &lt;i&gt;Saccharomyces cerevisiae&lt;/i&gt; 2μ plasmid&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;supports FLP-mediated excision with other FRT3 sites (&lt;a href=&quot;https://pubmed.ncbi.nlm.nih.gov/25605786/&quot;&gt;Bosch et al., 2015&lt;/a&gt;)&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="2" name="loxN" directionality="1" type="protein_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="1992-2025" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;Cre recombinase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;Cre-mediated recombination occurs in the 8-bp core sequence (AGGTATAC) (&lt;a href=&quot;https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8116776/&quot;&gt;Shaw et al., 2021&lt;/a&gt;). &lt;i&gt;loxN&lt;/i&gt; sites are compatible with each other, but incompatible with &lt;i&gt;loxP&lt;/i&gt; or &lt;i&gt;lox2272&lt;/i&gt; sites.&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="3" name="loxN" directionality="2" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="1500-1533" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;Cre recombinase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;Cre-mediated recombination occurs in the 8-bp core sequence (AGGTATAC) (&lt;a href=&quot;https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8116776/&quot;&gt;Shaw et al., 2021&lt;/a&gt;). &lt;i&gt;loxN&lt;/i&gt; sites are compatible with each other, but incompatible with &lt;i&gt;loxP&lt;/i&gt; or &lt;i&gt;lox2272&lt;/i&gt; sites.&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="4" name="lox2272" directionality="1" type="protein_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="1908-1941" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;Cre recombinase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;Cre-mediated recombination occurs in the 8-bp core sequence (AAGTATCC) (&lt;u&gt;&lt;a href=&quot;https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8116776/&quot;&gt;Shaw et al., 2021&lt;/u&gt;&lt;/a&gt;). &lt;i&gt;lox2272&lt;/i&gt; sites are compatible with each other, but incompatible with &lt;i&gt;loxP&lt;/i&gt; or &lt;i&gt;loxN&lt;/i&gt; sites (&lt;a href=&quot;https://www.ncbi.nlm.nih.gov/pubmed/9714735&quot;&gt;Lee and Saito, 1988&lt;/a&gt;).&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="5" name="lox2272" directionality="2" type="protein_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="1416-1449" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;Cre recombinase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;Cre-mediated recombination occurs in the 8-bp core sequence (AAGTATCC) (&lt;u&gt;&lt;a href=&quot;https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8116776/&quot;&gt;Shaw et al., 2021&lt;/u&gt;&lt;/a&gt;). &lt;i&gt;lox2272&lt;/i&gt; sites are compatible with each other, but incompatible with &lt;i&gt;loxP&lt;/i&gt; or &lt;i&gt;loxN&lt;/i&gt; sites (&lt;a href=&quot;https://www.ncbi.nlm.nih.gov/pubmed/9714735&quot;&gt;Lee and Saito, 1988&lt;/a&gt;).&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="6" name="Factor Xa site" directionality="2" translationMW="473.53" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="3213" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="3139" maxFusedRunOn="3139" detectionMode="exactProteinMatch"><Segment range="3214-3225" color="#cc99b2" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;Factor Xa recognition and cleavage site&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="IEGR"/></Q></Feature><Feature recentID="7" name="AAV2 ITR" type="repeat_region" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3879-4019" color="#ffe4c4" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;inverted terminal repeat of adeno-associated virus serotype 2&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="rpt_type"><V predef="inverted"/></Q></Feature><Feature recentID="8" name="WPRE" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2743-3331" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;woodchuck hepatitis virus&amp;nbsp;posttranscriptional regulatory element&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="hGH poly(A) signal" type="polyA_signal" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="3363-3839" color="#a6acb3" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;human growth hormone polyadenylation signal&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="10" name="EF-1α intron A" type="intron" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="462-1400" color="noColor" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;intron upstream of the start codon of human EF-1α&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="11" name="EF-1α promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="231-1409" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;strong constitutive promoter for human elongation factor EF-1α&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="Kozak sequence" type="regulatory" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="1540-1549" 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><Feature recentID="13" name="f1 ori" directionality="1" type="rep_origin" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4094-4549" color="#ffff00" type="standard"/><Q name="direction"><V text="RIGHT"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="ori" directionality="1" type="rep_origin" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5967-6555" 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="AmpR" directionality="1" translationMW="31557.36" type="CDS" allowSegmentOverlaps="0" cleavageArrows="5004" consecutiveTranslationNumbering="1" hitsStopCodon="1" detectionMode="exactProteinMatch"><Segment name="signal sequence" range="4936-5004" color="#ccffcc" type="standard" translated="1"/><Segment range="5005-5796" 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;β-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="16" name="AmpR promoter" directionality="1" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4831-4935" 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="17" name="AAV2 ITR (alternate)" type="repeat_region" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" prioritize="1" originalSequence="CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT"><Segment range="1-130" color="#ffcc99" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;Functional equivalent of wild-type AAV2 ITR&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="18" name="AAV2 ITR" type="repeat_region" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" prioritize="1" originalSequence="CTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT"><Segment range="3879-4008" color="#ffcc99" type="standard"/></Feature></Features>
  <?xml version="1.0"?><Primers nextValidID="12"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40" showAdditionalFivePrimeMatches="1" minimumFivePrimeAnnealing="15"/><Primer recentID="0" name="Amp-R" sequence="ATAATACCGCGCCACATAGC" description="Ampicillin resistance gene, reverse primer"><BindingSite location="5153-5172" boundStrand="1" annealedBases="ATAATACCGCGCCACATAGC" meltingTemperature="58"><Component hybridizedRange="5153-5172" bases="ATAATACCGCGCCACATAGC"/></BindingSite><BindingSite simplified="1" location="5153-5172" boundStrand="1" annealedBases="ATAATACCGCGCCACATAGC" meltingTemperature="58"><Component hybridizedRange="5153-5172" bases="ATAATACCGCGCCACATAGC"/></BindingSite></Primer><Primer recentID="1" name="EF1a-F" sequence="TCAAGCCTCAGACAGTGGTTC" description="Human elongation factor-1a promoter, forward primer"><BindingSite location="1356-1376" boundStrand="0" annealedBases="TCAAGCCTCAGACAGTGGTTC" meltingTemperature="58"><Component hybridizedRange="1356-1376" bases="TCAAGCCTCAGACAGTGGTTC"/></BindingSite><BindingSite simplified="1" location="1356-1376" boundStrand="0" annealedBases="TCAAGCCTCAGACAGTGGTTC" meltingTemperature="58"><Component hybridizedRange="1356-1376" bases="TCAAGCCTCAGACAGTGGTTC"/></BindingSite></Primer><Primer recentID="2" name="F1ori-F" sequence="GTGGACTCTTGTTCCAAACTGG" description="F1 origin, forward primer"><BindingSite location="4390-4411" boundStrand="0" annealedBases="GTGGACTCTTGTTCCAAACTGG" meltingTemperature="57"><Component hybridizedRange="4390-4411" bases="GTGGACTCTTGTTCCAAACTGG"/></BindingSite><BindingSite location="6307-6316" boundStrand="1" annealedBases="TCCAACTGG" meltingTemperature="13"><Component bases="GTGGACTCTTGT"/><Component hybridizedRange="6312-6316" bases="TCCAA"/><Component bases="A"/><Component hybridizedRange="6307-6310" bases="CTGG"/></BindingSite><BindingSite simplified="1" location="4390-4411" boundStrand="0" annealedBases="GTGGACTCTTGTTCCAAACTGG" meltingTemperature="57"><Component hybridizedRange="4390-4411" bases="GTGGACTCTTGTTCCAAACTGG"/></BindingSite><BindingSite simplified="1" location="6307-6316" boundStrand="1" annealedBases="TCCAACTGG" meltingTemperature="13"><Component bases="GTGGACTCTTGT"/><Component hybridizedRange="6312-6316" bases="TCCAA"/><Component bases="A"/><Component hybridizedRange="6307-6310" bases="CTGG"/></BindingSite></Primer><Primer recentID="3" name="F1ori-R" sequence="AGGGAAGAAAGCGAAAGGAG" description="F1 origin, reverse primer"><BindingSite location="4180-4199" boundStrand="1" annealedBases="AGGGAAGAAAGCGAAAGGAG" meltingTemperature="57"><Component hybridizedRange="4180-4199" bases="AGGGAAGAAAGCGAAAGGAG"/></BindingSite><BindingSite simplified="1" location="4180-4199" boundStrand="1" annealedBases="AGGGAAGAAAGCGAAAGGAG" meltingTemperature="57"><Component hybridizedRange="4180-4199" bases="AGGGAAGAAAGCGAAAGGAG"/></BindingSite></Primer><Primer recentID="4" name="hGH-PA-R" sequence="CCAGCTTGGTTCCCAATAGA" description="Human growth hormone terminator, reverse primer"><BindingSite location="3561-3580" boundStrand="1" annealedBases="CCAGCTTGGTTCCCAATAGA" meltingTemperature="55"><Component hybridizedRange="3561-3580" bases="CCAGCTTGGTTCCCAATAGA"/></BindingSite><BindingSite simplified="1" location="3561-3580" boundStrand="1" annealedBases="CCAGCTTGGTTCCCAATAGA" meltingTemperature="55"><Component hybridizedRange="3561-3580" bases="CCAGCTTGGTTCCCAATAGA"/></BindingSite></Primer><Primer recentID="5" name="mCherry-F" sequence="CCCCGTAATGCAGAAGAAGA" description="mCherry, forward primer"><BindingSite location="2435-2454" boundStrand="1" annealedBases="CCCCGTAATGCAGAAGAAGA" meltingTemperature="54"><Component hybridizedRange="2435-2454" bases="CCCCGTAATGCAGAAGAAGA"/></BindingSite><BindingSite simplified="1" location="2435-2454" boundStrand="1" annealedBases="CCCCGTAATGCAGAAGAAGA" meltingTemperature="54"><Component hybridizedRange="2435-2454" bases="CCCCGTAATGCAGAAGAAGA"/></BindingSite></Primer><Primer recentID="6" name="mCherry-R" sequence="TTGGTCACCTTCAGCTTGG" description="mCherry, reverse primer"><BindingSite location="1690-1708" boundStrand="1" annealedBases="TTGGTCACCTTCAGCTTGG" meltingTemperature="57"><Component hybridizedRange="1690-1708" bases="TTGGTCACCTTCAGCTTGG"/></BindingSite><BindingSite simplified="1" location="1690-1708" boundStrand="1" annealedBases="TTGGTCACCTTCAGCTTGG" meltingTemperature="57"><Component hybridizedRange="1690-1708" bases="TTGGTCACCTTCAGCTTGG"/></BindingSite></Primer><Primer recentID="7" name="pBR322ori-F" sequence="GGGAAACGCCTGGTATCTTT" description="pBR322 origin, forward primer"><BindingSite location="6455-6474" boundStrand="0" annealedBases="GGGAAACGCCTGGTATCTTT" meltingTemperature="56"><Component hybridizedRange="6455-6474" bases="GGGAAACGCCTGGTATCTTT"/></BindingSite><BindingSite simplified="1" location="6455-6474" boundStrand="0" annealedBases="GGGAAACGCCTGGTATCTTT" meltingTemperature="56"><Component hybridizedRange="6455-6474" bases="GGGAAACGCCTGGTATCTTT"/></BindingSite></Primer><Primer recentID="8" name="pBRforEco" sequence="AATAGGCGTATCACGAGGC" description="pBR322 vectors, upsteam of EcoRI site, forward primer"><BindingSite location="4744-4762" boundStrand="1" annealedBases="AATAGGCGTATCACGAGGC" meltingTemperature="57"><Component hybridizedRange="4744-4762" bases="AATAGGCGTATCACGAGGC"/></BindingSite><BindingSite simplified="1" location="4744-4762" boundStrand="1" annealedBases="AATAGGCGTATCACGAGGC" meltingTemperature="57"><Component hybridizedRange="4744-4762" bases="AATAGGCGTATCACGAGGC"/></BindingSite></Primer><Primer recentID="9" name="pGEX 3'" sequence="CCGGGAGCTGCATGTGTCAGAGG" description="pGEX vectors, reverse primer"><BindingSite location="4684-4706" boundStrand="0" annealedBases="CCGGGAGCTGCATGTGTCAGAGG" meltingTemperature="66"><Component hybridizedRange="4684-4706" bases="CCGGGAGCTGCATGTGTCAGAGG"/></BindingSite><BindingSite simplified="1" location="4684-4706" boundStrand="0" annealedBases="CCGGGAGCTGCATGTGTCAGAGG" meltingTemperature="66"><Component hybridizedRange="4684-4706" bases="CCGGGAGCTGCATGTGTCAGAGG"/></BindingSite></Primer><Primer recentID="10" name="pRS-marker" sequence="CGGCATCAGAGCAGATTGTA" description="pRS vectors, use to sequence yeast selectable marker"><BindingSite location="4565-4584" boundStrand="1" annealedBases="CGGCATCAGAGCAGATTGTA" meltingTemperature="54"><Component hybridizedRange="4565-4584" bases="CGGCATCAGAGCAGATTGTA"/></BindingSite><BindingSite simplified="1" location="4565-4584" boundStrand="1" annealedBases="CGGCATCAGAGCAGATTGTA" meltingTemperature="54"><Component hybridizedRange="4565-4584" bases="CGGCATCAGAGCAGATTGTA"/></BindingSite></Primer><Primer recentID="11" name="WPRE-R" sequence="CATAGCGTAAAAGGAGCAACA" description="WPRE, reverse primer"><BindingSite location="2795-2815" boundStrand="1" annealedBases="CATAGCGTAAAAGGAGCAACA" meltingTemperature="55"><Component hybridizedRange="2795-2815" bases="CATAGCGTAAAAGGAGCAACA"/></BindingSite><BindingSite simplified="1" location="2795-2815" boundStrand="1" annealedBases="CATAGCGTAAAAGGAGCAACA" meltingTemperature="55"><Component hybridizedRange="2795-2815" bases="CATAGCGTAAAAGGAGCAACA"/></BindingSite></Primer></Primers>
  ><Notes>
<UUID>a929cea2-a090-4d6e-bf20-04e40ee535b7</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<CustomMapLabel>pAAV-Ef1a-Coff/Fon-mCherry</CustomMapLabel>
<UseCustomMapLabel>1</UseCustomMapLabel>
<Description>Intersectional viral expression of mCherry in cells expressing Flp AND NOT Cre.</Description>
<Created UTC="5:0:0">2025.12.15</Created>
<LastModified UTC="5:0:0">2025.12.15</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
<References>
<Reference authors="Fenno LE, Ramakrishnan C, Kim YS, Evans KE, Lo M, Vesuna S, Inoue M, Cheung KYM, Yuen E, Pichamoorthy N, Hong ASO, Deisseroth K" journal="Neuron. 2020 Jun 20. pii: S0896-6273(20)30432-3. doi: 10.1016/j.neuron.2020.06.003." journalName="Neuron. 2020 Jun 20. pii: S0896-6273(20)30432-3. doi: 10.1016/j.neuron.2020.06.003." pubMedID="32574559" title="Comprehensive Dual- and Triple-Feature Intersectional Single-Vector Delivery of Diverse Functional Payloads to Cells of Behaving Mammals." type="Journal Article"/>
</References>
</Notes>
  Y              Unique 6+ Cutters                                                                                            d     THO  d              E              Y? P                                                                                                                                                           #   <7zXZ  ִF !   t/ []   S.( .~}    YZ   3<?xml version="1.0"?><EnzymeVisibilities vals=""/>
   )<?xml version="1.0"?><CustomEnzymeSets/>
