LOCUS       sequence-208670-       16366 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Target a Flp and Cre-dependent tdTomato expression cassette to the 
            mouse Rosa26 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16366)
  AUTHORS   Madisen L, Garner AR, Shimaoka D, Chuong AS, Klapoetke NC, Li L, van
            der Bourg A, Niino Y, Egolf L, Monetti C, Gu H, Mills M, Cheng A, 
            Tasic B, Nguyen TN, Sunkin SM, Benucci A, Nagy A, Miyawaki A, 
            Helmchen F, Empson RM, Knopfel T, Boyden ES, Reid RC, Carandini M, 
            Zeng H
  TITLE     Transgenic mice for intersectional targeting of neural sensors and 
            effectors with high specificity and performance.
  JOURNAL   Neuron. 2015 Mar 4;85(5):942-58. doi: 10.1016/j.neuron.2015.02.022.
  PUBMED    25741722
REFERENCE   2  (bases 1 to 16366)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.neuron.2015.02"; journalName: "Neuron"; date: "2015-03-4-
            4"; volume: "85"; issue: "5"; pages: "942-58"
COMMENT     Sequence Label: Ai65(RCFL-tdT) targeting vector
FEATURES             Location/Qualifiers
     source          1..16366
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    3..1085
                     /label=Rosa26 left arm
                     /note="left homology arm from intron 1 of the mouse Rosa26 
                     gene (Soriano, 1999)"
     enhancer        1181..1484
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1486..1763
                     /label=chicken beta-actin promoter
     intron          1764..2780
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2704..2726
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2788..2807
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    2862..2895
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     polyA_signal    2941..2989
                     /note="synthetic polyadenylation signal"
     polyA_signal    2998..3474
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3197..3216)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3509..3528)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3553..3601
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    3761..3794
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    3809..3842
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    4009..4143
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    4263..4397
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    4517..4651
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    4680..4713
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      4801..4810
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4807..6237
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     misc_feature    6316..6904
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6369..6389)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6787..6798)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6907..6923)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6908..6924)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(6925..6942)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6931..7138
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    7146..7179
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        7201..7700
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7236..7259)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7633..7652
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        7717..7764
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7792..8595
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7855..7874)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8465..8484
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8605..9066
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     misc_feature    9132..13472
                     /label=Rosa26 right arm
                     /note="right homology arm from intron 1 of the mouse Rosa26
                     gene (Soriano, 1999)"
     primer_bind     complement(13518..13538)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(13518..13536)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(13557..13573)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13557..13573)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13570..13592)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13581..13597
                     /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(13605..13611,13612..13629,13630..13635))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13650..13671
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13788..13805)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13959..14547)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14039..14058)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14718..15509,15510..15578))
                     /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     15341..15360
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15579..15683)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(15736..16164)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(15846..15867)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     16058..16077
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     16291..16313
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     16305..16322
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     16306..16322
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     16332..16351
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        16332..16350
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ccgcggcagg ccctccgagc gtggtggagc cgttctgtga gacagccggg tacgagtcgt
       61 gacgctggaa ggggcaagcg ggtggtgggc aggaatgcgg tccgccctgc agcaaccgga
      121 gggggaggga gaagggagcg gaaaagtctc caccggacgc ggccatggct cggggggggg
      181 ggggcagcgg aggagcgctt ccggccgacg tctcgtcgct gattggcttc ttttcctccc
      241 gccgtgtgtg aaaacacaaa tggcgtgttt tggttggcgt aaggcgcctg tcagttaacg
      301 gcagccggag tgcgcagccg ccggcagcct cgctctgccc actgggtggg gcgggaggta
      361 ggtggggtga ggcgagctgg acgtgcgggc gcggtcggcc tctggcgggg cgggggrggg
      421 gagggagggt cagcgaaagt agctcgcgcg cgagcggccg cccaccctcc ccttcctctg
      481 ggggagtcgt tttacccgcc gccggccggg cctcgtcgtc tgattggctc tcggggccca
      541 gaaaactggc ccttgccatt ggctcgtgtt cgtgcaagtt gagtccatcc gccggccagc
      601 gggggcggcg aggaggcgct cccaggttcc ggccctcccc tcggccccgc gccgcagagt
      661 ctggccgcgc gcccctgcgc aacgtggcag gaagcgcgcg ctgggggcgg ggacgggcag
      721 tagggctgag cggctgcggg gcgggtgcaa gcacgtttcc gacttgagtt gcctcaagag
      781 gggcgtgctg agccagacct ccatcgcgca ctccggggag tggagggaag gagcgagggc
      841 tcagttgggc tgttttggag gcaggaagca cttgctctcc caaagtcgct ctgagttgtt
      901 atcagtaagg gagctgcagt ggagtaggcg gggagaaggc cgcacccttc tccggagggg
      961 ggaggggagt gttgcaatac ctttctggga gttctctgct gcctcctggc ttctgaggac
     1021 cgccctgggc ctgggagaat cccttccccc tcttccctcg tgatctgcaa ctccagtctt
     1081 tctagcctta attaaccgtt taaacaattc tgcaggaatc tagttattaa tagtaatcaa
     1141 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     1201 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     1261 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     1321 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     1381 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     1441 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
     1501 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta
     1561 ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccrggsg
     1621 gggcggggsg gggsgrgggg sggggsgggg cgrggcggag aggtgcggcg gcagccaatc
     1681 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata
     1741 aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct
     1801 ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga
     1861 gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg
     1921 tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg
     1981 ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg
     2041 cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg
     2101 tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg cggggggggc tgcgagggga
     2161 acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg
     2221 tcgggctgca accccccctg cacccccctc cccgagttgc tgagcacggc ccggcttcgg
     2281 gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc
     2341 aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggagggctcg ggggaggggc
     2401 gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt
     2461 tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga
     2521 aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg
     2581 gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc
     2641 ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg
     2701 cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg
     2761 ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt
     2821 ttggcaaaga attgatttga taccgcgggc ccgtattcga agaagttcct attctctaga
     2881 aagtatagga acttccctca gcaccatggc tagcggcagc ctcggagttt gaatagatag
     2941 aataaaatat ctttattttc attccatctg tgtgttggtt ttttgtgtga gatctacggg
     3001 tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg
     3061 cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc
     3121 tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc
     3181 tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc
     3241 actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcaggctcc cgagttgttg
     3301 ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt
     3361 ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg
     3421 cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgcct
     3481 cagctaattg agtagggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     3541 cccgcgctat gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt
     3601 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     3661 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     3721 ggcccagggc tcgcagccaa cgtcggggcg gcaggccgtg gaagttccta ttctctagaa
     3781 agtataggaa cttcctcgag atttaaatat aacttcgtat aatgtatgct atacgaagtt
     3841 attcgcgatg aataaatgaa agcttgcaga tctgcgactc tagaggatct gcgactctag
     3901 aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     3961 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     4021 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4081 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     4141 atctgcgact ctagaggatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     4201 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     4261 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4321 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4381 cttatcatgt ctggatctgc gactctagag gatcataatc agccatacca catttgtaga
     4441 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     4501 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4561 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4621 actcatcaat gtatcttatc atgtctggat ccccatcaag ctgatccgga acccttaata
     4681 taacttcgta taatgtatgc tatacgaagt tattaggtcc ctcgacctgc agcccaagct
     4741 agatcgaatt cggccggcct tcacgtgttg cacttaacgc gttaagtgca acacgatccc
     4801 gccaccatgg tgagcaaggg cgaggaggtc atcaaagagt tcatgcgctt caaggtgcgc
     4861 atggagggct ccatgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     4921 tacgagggca cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc
     4981 tgggacatcc tgtcccccca gttcatgtac ggctccaagg cgtacgtgaa gcaccccgcc
     5041 gacatccccg attacaagaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     5101 aacttcgagg acggcggtct ggtgaccgtg acccaggact cctccctgca ggacggcacg
     5161 ctgatctaca aggtgaagat gcgcggcacc aacttccccc ccgacggccc cgtaatgcag
     5221 aagaagacca tgggctggga ggcctccacc gagcgcctgt acccccgcga cggcgtgctg
     5281 aagggcgaga tccaccaggc cctgaagctg aaggacggcg gccactacct ggtggagttc
     5341 aagaccatct acatggccaa gaagcccgtg caactgcccg gctactacta cgtggacacc
     5401 aagctggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgagcgctcc
     5461 gagggccgcc accacctgtt cctggggcat ggcaccggca gcaccggcag cggcagctcc
     5521 ggcaccgcct cctccgagga caacaacatg gccgtcatca aagagttcat gcgcttcaag
     5581 gtgcgcatgg agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc
     5641 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc
     5701 ttcgcctggg acatcctgtc cccccagttc atgtacggct ccaaggcgta cgtgaagcac
     5761 cccgccgaca tccccgatta caagaagctg tccttccccg agggcttcaa gtgggagcgc
     5821 gtgatgaact tcgaggacgg cggtctggtg accgtgaccc aggactcctc cctgcaggac
     5881 ggcacgctga tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta
     5941 atgcagaaga agaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc
     6001 gtgctgaagg gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg
     6061 gagttcaaga ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctactacgtg
     6121 gacaccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag
     6181 cgctccgagg gccgccacca cctgttcctg tacggcatgg acgagctgta caagtaagaa
     6241 ttgtgttgca cttaacgcgt taagtgcaac acgtgaaggc cggccctgca ggaattcgat
     6301 atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     6361 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     6421 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     6481 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     6541 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     6601 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     6661 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     6721 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     6781 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     6841 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     6901 ccgcatcgat accgtcgacc tcgacctcga ctgtgccttc tagttgccag ccatctgttg
     6961 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     7021 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     7081 gggtggggca ggacagcaag ggggaggatt gggaagacaa tggcaggcat gctggggaac
     7141 tagtggtgcc agggcgtgcc cttgggctcc ccgggcgcgg cggccgcatc gaattctacc
     7201 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg
     7261 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc
     7321 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca
     7381 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac
     7441 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt
     7501 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg
     7561 gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct
     7621 ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt
     7681 cctcatctcc gggcctttcg acctgcagca gcacgtgttg acaattaatc atcggcatag
     7741 tatatcggca tagtataata cgacaaggtg aggaactaaa ccgctagcac catgggatcg
     7801 gccattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     7861 ggctatgact gggcacaaca gacgatcggc tgctctgatg ccgccgtgtt ccggctgtca
     7921 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     7981 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     8041 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     8101 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     8161 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     8221 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     8281 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     8341 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     8401 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     8461 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     8521 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     8581 gacgagttct tctgagggga tccgctgtaa gtctgcagaa attgatgatc tattaaacaa
     8641 taaagatgtc cactaaaatg gaagtttttc ctgtcatact ttgttaagaa gggtgagaac
     8701 agagtaccta cattttgaat ggaaggattg gagctacggg ggtgggggtg gggtgggatt
     8761 agataaatgc ctgctcttta ctgaaggctc tttactattg ctttatgata atgtttcata
     8821 gttggatatc ataatttaaa caagcaaaac caaattaagg gccagctcat tcctcccact
     8881 catgatctat agatctatag atctctcgtg ggatcattgt ttttctcttg attcccactt
     8941 tgtggttcta agtactgtgg tttccaaatg tgtcagtttc atagcctgaa gaacgagatc
     9001 agcagcctct gttccacata cacttcattc tcagtattgt tttgccaagt tctaattcca
     9061 tcagaaagct tgcagaagat ctccccaact ggggtaacct ttgagttctc tcagttgggg
     9121 gggcgcgccg gctagaagat gggcgggagt cttctgggca ggcttaaagg ctaacctggt
     9181 gtgtgggcgt tgtcctgcag gggaattgaa caggtgtaaa attggaggga caagacttcc
     9241 cacagatttt cggttttgtc gggaagtttt ttaatagggg caaataagga aaatgggagg
     9301 ataggtagtc atctggggtt ttatgcagca aaactacagg ttattattgc ttgtgatccg
     9361 cctcggagta ttttccatcg aggtagatta aagacatgct cacccgagtt ttatactctc
     9421 ctgcttgaga tccttactac agtatgaaat tacagtgtcg cgagttagac tatgtaagca
     9481 gaattttaat catttttaaa gagcccagta cttcatatcc atttctcccg ctccttctgc
     9541 agccttatca aaaggtattt tagaacactc attttagccc cattttcatt tattatactg
     9601 gcttatccaa cccctagaca gagcattggc attttccctt tcctgatctt agaagtctga
     9661 tgactcatga aaccagacag attagttaca tacaccacaa atcgaggctg tagctggggc
     9721 ctcaacactg cagttctttt ataactcctt agtacacttt ttgttgatcc tttgccttga
     9781 tccttaattt tcagtgtcta tcacctctcc cgtcaggtgg tgttccacat ttgggcctat
     9841 tctcagtcca gggagtttta caacaataga tgtattgaga atccaaccta aagcttaact
     9901 ttccactccc atgaatgcct ctctcctttt tctccattta taaactgagc tattaaccat
     9961 taatggtttc caggtggatg tctcctcccc caatattacc tgatgtatct tacatattgc
    10021 caggctgata ttttaagaca ttaaaaggta tatttcatta ttgagccaca tggtattgat
    10081 tactgcttac taaaattttg tcattgtaca catctgtaaa aggtggttcc ttttggaatg
    10141 caaagttcag gtgtttgttg tctttcctga cctaaggtct tgtgagcttg tattttttct
    10201 atttaagcag tgctttctct tggactggct tgactcatgg cattctacac gttattgctg
    10261 gtctaaatgt gattttgcca agcttcttca ggacctataa ttttgcttga cttgtagcca
    10321 aacacaagta aaatgattaa gcaacaaatg tatttgtgaa gcttggtttt taggttgttg
    10381 tgttgtgtgt gcttgtgctc tataataata ctatccaggg gctggagagg tggctcggag
    10441 ttcaagagca cagactgctc ttccagaagt cctgagttca attcccagca accacatggt
    10501 ggctcacaac catctgtaat gggatctgat gccctcttct ggtgtgtctg aagaccacaa
    10561 gtgtattcac attaaataaa taaatcctcc ttcttcttct tttttttttt tttaaagaga
    10621 atactgtctc cagtagaatt tactgaagta atgaaatact ttgtgtttgt tccaatatgg
    10681 tagccaataa tcaaattact ctttaagcac tggaaatgtt accaaggaac taatttttat
    10741 ttgaagtgta actgtggaca gaggagccat aactgcagac ttgtgggata cagaagacca
    10801 atgcagactt taatgtcttt tctcttacac taagcaataa agaaataaaa attgaacttc
    10861 tagtatccta tttgtttaaa ctgctagctt tacttaactt ttgtgcttca tctatacaaa
    10921 gctgaaagct aagtctgcag ccattactaa acatgaaagc aagtaatgat aattttggat
    10981 ttcaaaaatg tagggccaga gtttagccag ccagtggtgg tgcttgcctt tatgccttta
    11041 atcccagcac tctggaggca gagacaggca gatctctgag tttgagccca gcctggtcta
    11101 cacatcaagt tctatctagg atagccagga atacacacag aaaccctgtt ggggaggggg
    11161 gctctgagat ttcataaaat tataattgaa gcattcccta atgagccact atggatgtgg
    11221 ctaaatccgt ctacctttct gatgagattt gggtattatt ttttctgtct ctgctgttgg
    11281 ttgggtcttt tgacactgtg ggctttcttt aaagcctcct tcctgccatg tggtctcttg
    11341 tttgctacta acttcccatg gcttaaatgg catggctttt tgccttctaa gggcagctgc
    11401 tgagatttgc agcctgattt ccagggtggg gttgggaaat ctttcaaaca ctaaaattgt
    11461 cctttaattt tttttttaaa aaatgggtta tataataaac ctcataaaat agttatgagg
    11521 agtgaggtgg actaatatta aatgagtccc tcccctataa aagagctatt aaggcttttt
    11581 gtcttatact taactttttt tttaaatgtg gtatctttag aaccaagggt cttagagttt
    11641 tagtatacag aaactgttgc atcgcttaat cagattttct agtttcaaat ccagagaatc
    11701 caaattcttc acagccaaag tcaaattaag aatttctgac ttttaatgtt aatttgctta
    11761 ctgtgaatat aaaaatgata gcttttcctg aggcagggtc tcactatgta tctctgcctg
    11821 atctgcaaca agatatgtag actaaagttc tgcctgcttt tgtctcctga atactaaggt
    11881 taaaatgtag taatactttt ggaacttgca ggtcagattc ttttataggg gacacactaa
    11941 gggagcttgg gtgatagttg gtaaaatgtg tttcaagtga tgaaaacttg aattattatc
    12001 accgcaacct actttttaaa aaaaaaagcc aggcctgtta gagcatgctt aagggatccc
    12061 taggacttgc tgagcacaca agagtagtta cttggcaggc tcctggtgag agcatatttc
    12121 aaaaaacaag gcagacaacc aagaaactac agttaaggtt acctgtcttt aaaccatctg
    12181 catatacaca gggatattaa aatattccaa ataatatttc attcaagttt tcccccatca
    12241 aattgggaca tggatttctc cggtgaatag gcagagttgg aaactaaaca aatgttggtt
    12301 ttgtgatttg tgaaattgtt ttcaagtgat agttaaagcc catgagatac agaacaaagc
    12361 tgctatttcg aggtctcttg gtttatactc agaagcactt ctttgggttt ccctgcacta
    12421 tcctgatcat gtgctaggcc taccttaggc tgattgttgt tcaaataaac ttaagtttcc
    12481 tgtcaggtga tgtcatatga tttcatatat caaggcaaaa catgttatat atgttaaaca
    12541 tttgtactta atgtgaaagt taggtctttg tgggtttgat ttttaatttt caaaacctga
    12601 gctaaataag tcatttttac atgtcttaca tttggtggaa ttgtataatt gtggtttgca
    12661 ggcaagactc tctgacctag taaccctacc tatagagcac tttgctgggt cacaagtcta
    12721 ggagtcaagc atttcacctt gaagttgaga cgttttgtta gtgtatacta gtttatatgt
    12781 tggaggacat gtttatccag aagatattca ggactatttt tgactgggct aaggaattga
    12841 ttctgattag cactgttagt gagcattgag tggcctttag gcttgaattg gagtcacttg
    12901 tatatctcaa ataatgctgg ccttttttaa aaagcccttg ttctttatca ccctgttttc
    12961 tacataattt ttgttcaaag aaatacttgt ttggatctcc ttttgacaac aatagcatgt
    13021 tttcaagcca tatttttttt cctttttttt tttttttttg gtttttcgag acagggtttc
    13081 tctgtatagc cctggctgtc ctggaactca ctttgtagac caggctggcc tcgaactcag
    13141 aaatccgcct gcctctgcct cctgagtgcc gggattaaag gcgtgcacca ccacgcctgg
    13201 ctaagttgga tattttgtta tataactata accaatacta actccactgg gtggattttt
    13261 aattcagtca gtagtcttaa gtggtcttta ttggcccttc attaaaatct actgttcact
    13321 ctaacagagg ctgttggtac tagtggcact taagcaactt cctacggata tactagcaga
    13381 ttaagggtca gggatagaaa ctagtctagc gttttgtata cctaccagct ttatactacc
    13441 ttgttctgat agaaatattt caggacatct agcttatcga taccgtcgag ggggggcccg
    13501 gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
    13561 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
    13621 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
    13681 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
    13741 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
    13801 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
    13861 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
    13921 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
    13981 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
    14041 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
    14101 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
    14161 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
    14221 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
    14281 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
    14341 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
    14401 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
    14461 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
    14521 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
    14581 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
    14641 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
    14701 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
    14761 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
    14821 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
    14881 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
    14941 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
    15001 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
    15061 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
    15121 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
    15181 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
    15241 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
    15301 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
    15361 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
    15421 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
    15481 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
    15541 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
    15601 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
    15661 aataaacaaa taggggttcc gcgcacaytt mscckraawg kssmayskra mttgtaagcg
    15721 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
    15781 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
    15841 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
    15901 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
    15961 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
    16021 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
    16081 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
    16141 ttaatgcgcc gctacagggc gcgtcmsrtk ssmcwttymg gskgmgyrac tgttgggaag
    16201 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
    16261 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
    16321 gtgagcgcgc gtaatacgac tcactatagg gcgaattgga gctcca
//
