LOCUS       sequence-221186-       18313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Target a Cre-dependent ChrimsonR-tdTomato cassette and a tTA2 
            cassette into the mouse TIGRE locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18313)
  AUTHORS   Daigle TL, Madisen L, Hage TA, Valley MT, Knoblich U, Larsen RS, 
            Takeno MM, Huang L, Gu H, Larsen R, Mills M, Bosma-Moody A, Siverts 
            LA, Walker M, Graybuck LT, Yao Z, Fong O, Nguyen TN, Garren E, Lenz 
            GH, Chavarha M, Pendergraft J, Harrington J, Hirokawa KE, Harris JA,
            Nicovich PR, McGraw MJ, Ollerenshaw DR, Smith KA, Baker CA, Ting JT,
            Sunkin SM, Lecoq J, Lin MZ, Boyden ES, Murphy GJ, da Costa NM, 
            Waters J, Li L, Tasic B, Zeng H
  TITLE     A Suite of Transgenic Driver and Reporter Mouse Lines with Enhanced 
            Brain-Cell-Type Targeting and Functionality.
  JOURNAL   Cell. 2018 Jul 12;174(2):465-480.e22. doi: 
            10.1016/j.cell.2018.06.035.
  PUBMED    30007418
REFERENCE   2  (bases 1 to 18313)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2018.06"; journalName: "Cell"; date: "2018-07-12- 
            12"; volume: "174"; issue: "2"; pages: "465-480.e22"
COMMENT     Sequence Label: Ai167 (TIT2L-ChrimsonR-tdT-ICL-tTA2) targeting...
FEATURES             Location/Qualifiers
     source          1..18313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    414..447
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     polyA_signal    2959..3040
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2984..3003
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    join(3054..3072,3073..3095,3096..3114,3115..3137,3138..3156,
                     3157..3179,3180..3198,3199..3221,3222..3240,3241..3263,
                     3264..3282,3283..3305,3306..3324)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        3357..3395
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3392..3416
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    complement(3529..3562)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3640..3688
                     /note="synthetic polyadenylation signal"
     polyA_signal    3697..4173
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     polyA_signal    4185..4646
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     protein_bind    complement(4673..4706)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      4717..4726
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4723..5772
                     /codon_start=1
                     /product="red light-drivable channelrhodopsin from 
                     Chlamydomonas noctigama (Klapoetke et al., 2014)"
                     /label=ChrimsonR
                     /note="K176R mutant with accelerated closing kinetics"
                     /translation="MAELISSATRSLFAAGGINPWPNPYHHEDMGCGGMTPTGECFSTE
                     WWCDPSYGLSDAGYGYCFVEATGGYLVVGVEKKQAWLHSRGTPGEKIGAQVCQWIAFSI
                     AIALLTFYGFSAWKATCGWEEVYVCCVEVLFVTLEIFKEFSSPATVYLSTGNHAYCLRY
                     FEWLLSCPVILIRLSNLSGLKNDYSKRTMGLIVSCVGMIVFGMAAGLATDWLKWLLYIV
                     SCIYGGYMYFQAAKCYVEANHSVPKGHCRMVVKLMAYAYFASWGSYPILWAVGPEGLLK
                     LSPYANSIGHSICDIIAKEFWTFLAHHLRIKIHEHILIHGDIRKTTKMEIGGEEVEVEE
                     FVEEEDEDTV"
     misc_feature    7243..7831
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(7714..7725)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    7862..8069
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        10599..10902
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        10904..11181
                     /label=chicken beta-actin promoter
     intron          11183..12200
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     12124..12146
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     12208..12227
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    12288..12321
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     polyA_signal    12359..12407
                     /note="synthetic polyadenylation signal"
     polyA_signal    12416..12892
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(12927..12946)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    12971..13019
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    13198..13231
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             join(13256..13873,13874..13879,13880..13915,13916..13918,
                     13919..13954,13955..13957,13958..13993,13994..14002)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=tTA-Advanced
                     /note="In the Tet-Off(R) system, tTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    14045..14633
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(14516..14527)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    14664..14871
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    14885..14918
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        14945..15444
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(14980..15003)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     15377..15396
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     complement(16092..16108)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(16092..16108)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(16105..16127)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    16116..16132
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(16140..16146,16147..16164,16165..16170))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    16185..16206
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(16323..16340)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(16494..17082)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(16574..16593)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(17253..18044,18045..18113))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     17876..17895
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(18114..18218)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     18286..18304
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt accgaagttc
      421 ctattcttca aatagtatag gaacttcttt cggccggcca atcgattgga tccgtttaat
      481 tcgagctcgc tagagcccca tcctcactga ctccgtcctg gagttggatg agagataatg
      541 gccttacgtt gtgccagggg agggtcgggc tggatttagc aagatttacc ttctccaaag
      601 agcggtgctg cagtggcaca gctgcccacg gaggtggggg ggtcaccgtc cctggaggtg
      661 atgaagaact gtggggatgt ggcactgagg gacatggcca gtgggcacgg tgggtgggtt
      721 ggggttggtc ttggggatct tggagggctt ttccagcctt catgatttga cgattgtatg
      781 aacatctaca tggcaattct ccagctgcct gtcccagtcc tactgaccca gctgtatctc
      841 tccaggcaag ctcttccacc ccttctgctt gcatccagac accatcaaac atgcaggctc
      901 agacacaggg accagcagtg tctgtggcct ttttgtgctc ctctccatgc tgggttttaa
      961 cttgctcttt gtccttctat cctatcttct tatccttaag gctgttctga acgctgtgac
     1021 ttggagagtg tcccagagcc ctcaacacct gcatgtccca cgtccatgct gtcctgcact
     1081 tccttatccc caagatctgc ctctccgtga tgcactgaat tggcaaacat gtgtcacccc
     1141 agaccaacaa tgtcacagca aactccccct tgataggaca agggggaatg gctttacact
     1201 gagacagggg aggtttgggt tggatatgag gaggcagttt ttcccccaga gggtggtgac
     1261 gcactgaaca ggttgcccaa ggaggctgtg gatgccccat ccctgcaggc attcaaggcc
     1321 aggctggatg tggctctggg cagcctgggc tgctggttga tgaccctgca catagcaggg
     1381 ggttggatct ggatgagcac tgtgctcctt tgcaacccag gccgttctat gattctgtca
     1441 ttctaaatct ctctttcagc ctaaagcttt ttccccgtat ccccccaggt gtctgcaggc
     1501 tcaaagagca gcgagaagcg ttcagaggaa agcgatcccg tgccaccttc cccgtgcccg
     1561 ggctgtcccc gcacgctgcc ggctcgggga tgcgggggga gcgccggacc ggagcggagc
     1621 cccgggcggc tcgctgctgc cccctagcgg gggagggacg taattacatc cctgggggct
     1681 ttgggggggg gctgtcccga gctcggtacc gccccatcct cactgactcc gtcctggagt
     1741 tggatgagag ataatggcct tacgttgtgc caggggaggg tcgggctgga tttagcaaga
     1801 tttaccttct ccaaagagcg gtgctgcagt ggcacagctg cccacggagg tgggggggtc
     1861 accgtccctg gaggtgatga agaactgtgg ggatgtggca ctgagggaca tggccagtgg
     1921 gcacggtggg tgggttgggg ttggtcttgg ggatcttgga gggcttttcc agccttcatg
     1981 atttgacgat tgtatgaaca tctacatggc aattctccag ctgcctgtcc cagtcctact
     2041 gacccagctg tatctctcca ggcaagctct tccacccctt ctgcttgcat ccagacacca
     2101 tcaaacatgc aggctcagac acagggacca gcagtgtctg tggccttttt gtgctcctct
     2161 ccatgctggg ttttaacttg ctctttgtcc ttctatccta tcttcttatc cttaaggctg
     2221 ttctgaacgc tgtgacttgg agagtgtccc agagccctca acacctgcat gtcccacgtc
     2281 catgctgtcc tgcacttcct tatccccaag atctgcctct ccgtgatgca ctgaattggc
     2341 aaacatgtgt caccccagac caacaatgtc acagcaaact cccccttgat aggacaaggg
     2401 ggaatggctt tacactgaga caggggaggt ttgggttgga tatgaggagg cagtttttcc
     2461 cccagagggt ggtgacgcac tgaacaggtt gcccaaggag gctgtggatg ccccatccct
     2521 gcaggcattc aaggccaggc tggatgtggc tctgggcagc ctgggctgct ggttgatgac
     2581 cctgcacata gcagggggtt ggatctggat gagcactgtg ctcctttgca acccaggccg
     2641 ttctatgatt ctgtcattct aaatctctct ttcagcctaa agctttttcc ccgtatcccc
     2701 ccaggtgtct gcaggctcaa agagcagcga gaagcgttca gaggaaagcg atcccgtgcc
     2761 accttccccg tgcccgggct gtccccgcac gctgccggct cggggatgcg gggggagcgc
     2821 cggaccggag cggagccccg ggcggctcgc tgctgccccc tagcggggga gggacgtaat
     2881 tacatccctg ggggctttgg gggggggctg tcccggtacc cggggatcca ccggtatcga
     2941 ttcggccggc cgacgtcggc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3001 tgctttattt gtaaccatta taagctgcaa taaacaagtt tcgagtttac cactccctat
     3061 cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg
     3121 aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact
     3181 ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga
     3241 aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagttt
     3301 accactccct atcagtgata gagaaaagtg aaagtcgagc tcggtacccg ggtcgaggta
     3361 ggcgtgtacg gtgggaggcc tatataagca gagctcgttt agtgaaccgt cagatcgcct
     3421 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc
     3481 gcggccccga attcgagctc ggtacccggg gatcaaattc aattaaatat aacttcgtat
     3541 agcatacatt atacgaagtt attcgcgagt gaataaatga aagcttgcag atcccctcag
     3601 caccatggct agcggcagcc tcggagtttg aatagataga ataaaatatc tttattttca
     3661 ttccatctgt gtgttggttt tttgtgtgag atctacgggt ggcatccctg tgacccctcc
     3721 ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata
     3781 aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg
     3841 ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat
     3901 tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg
     3961 gttcaagcga ttctcctgcc tcaggctccc gagttgttgg gattccaggc atgcatgacc
     4021 aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg
     4081 gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta
     4141 caggcgtgaa ccactgctcc cttccctgtc cttctgcctc agcgctgtaa gtctgcagaa
     4201 attgatgatc tattaaacaa taaagatgtc cactaaaatg gaagtttttc ctgtcatact
     4261 ttgttaagaa gggtgagaac agagtaccta cattttgaat ggaaggattg gagctacggg
     4321 ggtgggggtg gggtgggatt agataaatgc ctgctcttta ctgaaggctc tttactattg
     4381 ctttatgata atgtttcata gttggatatc ataatttaaa caagcaaaac caaattaagg
     4441 gccagctcat tcctcccact catgatctat agatctatag atctctcgtg ggatcattgt
     4501 ttttctcttg attcccactt tgtggttcta agtactgtgg tttccaaatg tgtcagtttc
     4561 atagcctgaa gaacgagatc agcagcctct gttccacata cacttcattc tcagtattgt
     4621 tttgccaagt tctaattcca tcagaaagct tttaattaac tgaatagcta acataacttc
     4681 gtatagcata cattatacga agttatcacg cgtgccgcca ccatggctga gctgatcagc
     4741 agcgccacca gatctctgtt tgccgccgga ggcatcaacc cttggcctaa cccctaccac
     4801 cacgaggaca tgggctgtgg aggaatgaca cctacaggcg agtgcttcag caccgagtgg
     4861 tggtgtgacc cttcttacgg actgagcgac gccggatacg gatattgctt cgtggaggcc
     4921 acaggcggct acctggtcgt gggagtggag aagaagcagg cttggctgca cagcagaggc
     4981 acaccaggag aaaagatcgg cgcccaggtc tgccagtgga ttgctttcag catcgccatc
     5041 gccctgctga cattctacgg cttcagcgcc tggaaggcca cttgcggttg ggaggaggtc
     5101 tacgtctgtt gcgtcgaggt gctgttcgtg accctggaga tcttcaagga gttcagcagc
     5161 cccgccacag tgtacctgtc taccggcaac cacgcctatt gcctgcgcta cttcgagtgg
     5221 ctgctgtctt gccccgtgat cctgatcaga ctgagcaacc tgagcggcct gaagaacgac
     5281 tacagcaagc ggaccatggg cctgatcgtg tcttgcgtgg gaatgatcgt gttcggcatg
     5341 gccgcaggac tggctaccga ttggctcaag tggctgctgt atatcgtgtc ttgcatctac
     5401 ggcggctaca tgtacttcca ggccgccaag tgctacgtgg aagccaacca cagcgtgcct
     5461 aaaggccatt gccgcatggt cgtgaagctg atggcctacg cttacttcgc ctcttggggc
     5521 agctacccaa tcctctgggc agtgggacca gaaggactgc tgaagctgag cccttacgcc
     5581 aacagcatcg gccacagcat ctgcgacatc atcgccaagg agttttggac cttcctggcc
     5641 caccacctga ggatcaagat ccacgagcac atcctgatcc acggcgacat ccggaagacc
     5701 accaagatgg agatcggagg cgaggaggtg gaagtggaag agttcgtgga ggaggaggac
     5761 gaggacacag tggcggcacc ggtagtagca gtgagtaagg gcgaggaagt gatcaaagag
     5821 ttcatgcggt ttaaggtgag aatggaagga agcatgaacg gccacgagtt cgaaattgag
     5881 ggagaaggag agggacggcc ctacgagggc acccagacag ccaagctgaa agtgacaaag
     5941 ggcgggcctc tgccattcgc ttgggacatc ctgagcccac agtttatgta cggctccaag
     6001 gcctatgtga aacatccagc tgacattccc gattataaga aactgagctt ccccgagggg
     6061 tttaagtggg aaagagtgat gaacttcgag gacggaggcc tggtgactgt gacccaggac
     6121 agctccctgc aggatgggac cctgatctac aaggtgaaaa tgagagggac aaattttccc
     6181 cctgatggac ctgtgatgca gaagaaaact atgggatggg aggcctccac cgaaaggctg
     6241 tatccacgcg acggggtgct gaaaggagaa atccaccagg ctctgaagct gaaagatggg
     6301 ggacattacc tggtggagtt caagacaatc tacatggcca agaaacctgt gcagctgcca
     6361 ggctactatt acgtggacac aaaactggat atcacttcac acaacgagga ctacactatt
     6421 gtggagcagt atgaacggag cgaggggaga caccatctgt tcctgggcca tgggactgga
     6481 agtaccggct cagggtctag tggaaccgcc tcaagcgagg ataacaatat ggctgtgatc
     6541 aaagagttca tgaggtttaa ggtgcgcatg gagggcagca tgaatgggca cgaatttgag
     6601 attgaaggag agggcgaagg gaggccttac gagggcacac agactgccaa gctgaaagtg
     6661 accaagggag gaccactgcc tttcgcttgg gatatcctgt ctcctcagtt tatgtacgga
     6721 agtaaggcct atgtcaagca tcccgctgac attcctgatt acaagaaact gtctttccca
     6781 gagggcttta agtgggagag agtgatgaat tttgaagatg gaggcctggt gaccgtgaca
     6841 caggactcct ctctgcagga tggcactctg atctacaaag tcaaaatgcg cggcaccaat
     6901 tttccacccg atgggcccgt gatgcagaag aaaacaatgg ggtgggaggc cagcactgaa
     6961 cggctgtatc ctagagacgg agtgctgaag ggcgaaatcc accaggccct gaagctgaaa
     7021 gacggcggcc actacctggt ggagttcaaa accatctaca tggccaagaa accagtgcag
     7081 ctgcccggct attactatgt ggacaccaag ctggatatca catcccacaa tgaagactac
     7141 accattgtgg aacagtatga gaggtctgaa ggacgccacc atctgtttct gtacggcatg
     7201 gatgagctgt ataagtaaac gcgttacgta aagcttgtcg ataatcaacc tctggattac
     7261 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     7321 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     7381 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     7441 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     7501 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     7561 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     7621 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg
     7681 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     7741 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     7801 agtcggatct ccctttgggc cgcctccccg catcgacacc gagcgctgct cgaggcctcg
     7861 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     7921 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     7981 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     8041 tgggaagaca atagcaggca tgctggggac acgtgggatc ctctagtcta gagccccatc
     8101 ctcactgact ccgtcctgga gttggatgag agataatggc cttacgttgt gccaggggag
     8161 ggtcgggctg gatttagcaa gatttacctt ctccaaagag cggtgctgca gtggcacagc
     8221 tgcccacgga ggtggggggg tcaccgtccc tggaggtgat gaagaactgt ggggatgtgg
     8281 cactgaggga catggccagt gggcacggtg ggtgggttgg ggttggtctt ggggatcttg
     8341 gagggctttt ccagccttca tgatttgacg attgtatgaa catctacatg gcaattctcc
     8401 agctgcctgt cccagtccta ctgacccagc tgtatctctc caggcaagct cttccacccc
     8461 ttctgcttgc atccagacac catcaaacat gcaggctcag acacagggac cagcagtgtc
     8521 tgtggccttt ttgtgctcct ctccatgctg ggttttaact tgctctttgt ccttctatcc
     8581 tatcttctta tccttaaggc tgttctgaac gctgtgactt ggagagtgtc ccagagccct
     8641 caacacctgc atgtcccacg tccatgctgt cctgcacttc cttatcccca agatctgcct
     8701 ctccgtgatg cactgaattg gcaaacatgt gtcaccccag accaacaatg tcacagcaaa
     8761 ctcccccttg ataggacaag ggggaatggc tttacactga gacaggggag gtttgggttg
     8821 gatatgagga ggcagttttt cccccagagg gtggtgacgc actgaacagg ttgcccaagg
     8881 aggctgtgga tgccccatcc ctgcaggcat tcaaggccag gctggatgtg gctctgggca
     8941 gcctgggctg ctggttgatg accctgcaca tagcaggggg ttggatctgg atgagcactg
     9001 tgctcctttg caacccaggc cgttctatga ttctgtcatt ctaaatctct ctttcagcct
     9061 aaagcttttt ccccgtatcc ccccaggtgt ctgcaggctc aaagagcagc gagaagcgtt
     9121 cagaggaaag cgatcccgtg ccaccttccc cgtgcccggg ctgtccccgc acgctgccgg
     9181 ctcggggatg cggggggagc gccggaccgg agcggagccc cgggcggctc gctgctgccc
     9241 cctagcgggg gagggacgta attacatccc tgggggcttt gggggggggc tgtccctcta
     9301 gagccccatc ctcactgact ccgtcctgga gttggatgag agataatggc cttacgttgt
     9361 gccaggggag ggtcgggctg gatttagcaa gatttacctt ctccaaagag cggtgctgca
     9421 gtggcacagc tgcccacgga ggtggggggg tcaccgtccc tggaggtgat gaagaactgt
     9481 ggggatgtgg cactgaggga catggccagt gggcacggtg ggtgggttgg ggttggtctt
     9541 ggggatcttg gagggctttt ccagccttca tgatttgacg attgtatgaa catctacatg
     9601 gcaattctcc agctgcctgt cccagtccta ctgacccagc tgtatctctc caggcaagct
     9661 cttccacccc ttctgcttgc atccagacac catcaaacat gcaggctcag acacagggac
     9721 cagcagtgtc tgtggccttt ttgtgctcct ctccatgctg ggttttaact tgctctttgt
     9781 ccttctatcc tatcttctta tccttaaggc tgttctgaac gctgtgactt ggagagtgtc
     9841 ccagagccct caacacctgc atgtcccacg tccatgctgt cctgcacttc cttatcccca
     9901 agatctgcct ctccgtgatg cactgaattg gcaaacatgt gtcaccccag accaacaatg
     9961 tcacagcaaa ctcccccttg ataggacaag ggggaatggc tttacactga gacaggggag
    10021 gtttgggttg gatatgagga ggcagttttt cccccagagg gtggtgacgc actgaacagg
    10081 ttgcccaagg aggctgtgga tgccccatcc ctgcaggcat tcaaggccag gctggatgtg
    10141 gctctgggca gcctgggctg ctggttgatg accctgcaca tagcaggggg ttggatctgg
    10201 atgagcactg tgctcctttg caacccaggc cgttctatga ttctgtcatt ctaaatctct
    10261 ctttcagcct aaagcttttt ccccgtatcc ccccaggtgt ctgcaggctc aaagagcagc
    10321 gagaagcgtt cagaggaaag cgatcccgtg ccaccttccc cgtgcccggg ctgtccccgc
    10381 acgctgccgg ctcggggatg cggggggagc gccggaccgg agcggagccc cgggcggctc
    10441 gctgctgccc cctagcgggg gagggacgta attacatccc tgggggcttt gggggggggc
    10501 tgtccctcta gaagtcgacg gcgcgccctg caggaatcta gttattaata gtaatcaatt
    10561 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
    10621 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
    10681 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
    10741 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
    10801 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
    10861 acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg
    10921 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
    10981 ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggcgcgc gccaggcggg
    11041 gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc agccaatcag
    11101 agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa
    11161 aagcgaagcg cgcggcgggc ggggagtcgc tgcgacgctg ccttcgcccc gtgccccgct
    11221 ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga
    11281 gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg
    11341 tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg
    11401 ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg
    11461 cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg
    11521 tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg cggggggggc tgcgagggga
    11581 acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg
    11641 tcgggctgca accccccctg cacccccctc cccgagttgc tgagcacggc ccggcttcgg
    11701 gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc
    11761 aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggagggctcg ggggaggggc
    11821 gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt
    11881 tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga
    11941 aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg
    12001 gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc
    12061 ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg
    12121 cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg
    12181 ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt
    12241 ttggcaaaga attgatttga taccgcgggc cccgcgtgga cggtaccata acttcgtata
    12301 aagtatccta tacgaagtta tccatggcta gcggcagcct cggagtttga atagatagaa
    12361 taaaatatct ttattttcat tccatctgtg tgttggtttt ttgtgtgaga tctacgggtg
    12421 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
    12481 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
    12541 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
    12601 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
    12661 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct caggctcccg agttgttggg
    12721 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt
    12781 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc
    12841 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgcctca
    12901 gctaattgag taggggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
    12961 cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca
    13021 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
    13081 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
    13141 cccagggctc gcagccaacg tcggggcggc aggccgtgga agttcctatt ctctagaata
    13201 acttcgtata aagtatccta tacgaagtta tacgcgcgcg tccatttaat tcaccatgtc
    13261 tagactggac aagagcaaag tcataaactc tgctctggaa ttactcaatg aagtcggtat
    13321 cgaaggcctg acgacaagga aactcgctca aaagctggga gttgagcagc ctaccctgta
    13381 ctggcacgtg aagaacaagc gggccctgct cgatgccctg gcaatcgaga tgctggacag
    13441 gcatcatacc cacttctgcc ccctggaagg cgagtcatgg caagactttc tgcggaacaa
    13501 cgccaagtca ttccgctgtg ctctcctctc acatcgcgac ggggctaaag tgcatctcgg
    13561 cacccgccca acagagaaac agtacgaaac cctggaaaat cagctcgcgt tcctgtgtca
    13621 gcaaggcttc tccctggaga acgcactgta cgctctgtcc gccgtgggcc actttacact
    13681 gggctgcgta ttggaggatc aggagcatca agtagcaaaa gaggaaagag agacacctac
    13741 caccgattct atgcccccac ttctgagaca agcaattgag ctgttcgacc atcagggagc
    13801 cgaacctgcc ttccttttcg gcctggaact aatcatatgt ggcctggaga aacagctaaa
    13861 gtgcgaaagc ggcgggccgg ccgacgccct tgacgatttt gacttagaca tgctcccagc
    13921 cgatgccctt gacgactttg accttgatat gctgcctgct gacgctcttg acgattttga
    13981 ccttgacatg ctccccgggt aactaagtaa ggatcaaatg gcgcgttacg taaagcttgt
    14041 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
    14101 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
    14161 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
    14221 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
    14281 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
    14341 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
    14401 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
    14461 gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
    14521 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
    14581 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgaca
    14641 ccgagcgctg ctcgaggcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
    14701 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
    14761 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
    14821 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg acacgtggga
    14881 tccggtgcca gggcgtgccc ttgggctccc cgggcgcgtt tgggcggccg catcgaattc
    14941 taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg
    15001 ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt
    15061 aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta
    15121 gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta
    15181 gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc
    15241 ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga
    15301 aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg
    15361 tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc
    15421 tcttcctcat ctccgggcct ttcgacctgc aatcgccgct agccgccacc atgaaaaagc
    15481 ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg
    15541 acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc
    15601 gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt
    15661 atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca
    15721 gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc
    15781 ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg
    15841 cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat
    15901 acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa
    15961 ctgtgatgga cgacaccgtc agtgtaagtg ctagccccta ggcctgaatc gaagttccta
    16021 ttcttcaaaa ggtataggaa cttcgagctc gtcgactgca gaggcctgca tgcaagcttg
    16081 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
    16141 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
    16201 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
    16261 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
    16321 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
    16381 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
    16441 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
    16501 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
    16561 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
    16621 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
    16681 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
    16741 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
    16801 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
    16861 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
    16921 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
    16981 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
    17041 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
    17101 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
    17161 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
    17221 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
    17281 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
    17341 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
    17401 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
    17461 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
    17521 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
    17581 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
    17641 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
    17701 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
    17761 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
    17821 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
    17881 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
    17941 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
    18001 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
    18061 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
    18121 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
    18181 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
    18241 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
    18301 aggccctttc gtc
//
