LOCUS       sequence-100362-       15455 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Target a Cre-dependent GCaMP6-fast expression cassette to the mouse 
            Rosa26 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15455)
  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 15455)
  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: Ai95(RCL-GCaMP6f) targeting vector
FEATURES             Location/Qualifiers
     source          1..15455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    3..1087
                     /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          1765..2782
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2706..2728
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2790..2809
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(2844..2877)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    2955..3003
                     /note="synthetic polyadenylation signal"
     polyA_signal    3012..3488
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3211..3230)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     polyA_signal    3500..3961
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     protein_bind    complement(3988..4021)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      4047..4056
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4053..4061,4062..4079,4080..4082,4083..4115,4116..4118,
                     4119..4142,4143..4169,4170..4226,4227..4232,4233..4502,
                     4503..4520,4521..4955,4956..4961,4962..5405)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      4149..4158
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     4439..4460
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4566..4587)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4827..4846)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    5485..6073
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5538..5558)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5956..5967)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6076..6092)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6077..6093)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(6088..6105)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6094..6301
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    6309..6342
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        6364..6863
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6399..6422)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6796..6815
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     promoter        6880..6927
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6955..7758
                     /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(7018..7037)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7628..7647
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7768..8229
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     misc_feature    8293..12562
                     /label=Rosa26 right arm
                     /note="right homology arm from intron 1 of the mouse Rosa26
                     gene (Soriano, 1999)"
     primer_bind     complement(12607..12627)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(12607..12625)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(12646..12662)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12646..12662)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12659..12681)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12670..12686
                     /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(12694..12700,12701..12718,12719..12724))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12739..12760
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12877..12894)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13048..13636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13128..13147)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13807..14598,14599..14667))
                     /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     14430..14449
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14668..14772)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(14798..15253)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(14935..14956)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     15147..15166
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     15380..15402
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     15394..15411
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     15395..15411
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     15421..15440
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        15421..15439
                     /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 aggascgctt ccggccgacg tctcgtcgct gattggctty ttttcctccc
      241 gccgtgtgtg aaaacacaaa tggcgtgttt tggttggcgt aaggcgcctg tcagttaacg
      301 gcagccggag tgcgcagccg ccggcagcct cgctctgccc actgggtggg gcgggaggta
      361 ggtggggtga ggcgagctgn acgtgcgggc gcggtcggcc tctggcgggg cgggggaggg
      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 cccttgcccc ctcttcccct cgtgatctgc aactccagtc
     1081 tttctagtta 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 gcgccaggcg
     1621 gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc
     1681 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata
     1741 aaaagcgaag cgcgcggcgg gcggggagtc gctgcgacgc tgccttcgcc ccgtgccccg
     1801 ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt
     1861 gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct
     1921 tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg
     1981 ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc
     2041 cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag
     2101 tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg
     2161 gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc
     2221 ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg gcccggcttc
     2281 gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg
     2341 gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct cgggggaggg
     2401 gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca gccattgcct
     2461 tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct gtgcggagcc
     2521 gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc
     2581 cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct
     2641 ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg gacggggcag
     2701 ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca
     2761 tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca
     2821 ttttggcaaa gaattttcga aatataactt cgtatagcat acattatacg aagttattcg
     2881 cgagtgaata aatgaaagct tgcagatccc ctcagcacca tggctagcgg cagcctcgga
     2941 gtttgaatag atagaataaa atatctttat tttcattcca tctgtgtgtt ggttttttgt
     3001 gtgagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag
     3061 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg
     3121 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag
     3181 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg
     3241 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagg
     3301 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt
     3361 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga
     3421 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc
     3481 ctgtccttct gcctcagcgc tgtaagtctg cagaaattga tgatctatta aacaataaag
     3541 atgtccacta aaatggaagt ttttcctgtc atactttgtt aagaagggtg agaacagagt
     3601 acctacattt tgaatggaag gattggagct acgggggtgg gggtggggtg ggattagata
     3661 aatgcctgct ctttactgaa ggctctttac tattgcttta tgataatgtt tcatagttgg
     3721 atatcataat ttaaacaagc aaaaccaaat taagggccag ctcattcctc ccactcatga
     3781 tctatagatc tatagatctc tcgtgggatc attgtttttc tcttgattcc cactttgtgg
     3841 ttctaagtac tgtggtttcc aaatgtgtca gtttcatagc ctgaagaacg agatcagcag
     3901 cctctgttcc acatacactt cattctcagt attgttttgc caagttctaa ttccatcaga
     3961 aagcttttaa ttaactgaat agctaacata acttcgtata gcatacatta tacgaagtta
     4021 tcacgcgcca tttgggaacc ggtgccgcca ccatgggttc tcatcatcat catcatcatg
     4081 gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac gatgacgata
     4141 aggatctcgc caccatggtc gactcatcac gtcgtaagtg gaataagaca ggtcacgcag
     4201 tcagagctat aggtcggctg agctcactcg agaacgtcta tatcaaggcc gacaagcaga
     4261 agaacggcat caaggcgaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc
     4321 tcgcctacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     4381 accactacct gagcgtgcag tccaaacttt cgaaagaccc caacgagaag cgcgatcaca
     4441 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     4501 agggcggtac cggagggagc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     4561 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     4621 gtgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     4681 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     4741 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctaca
     4801 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     4861 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     4921 acggcaacat cctggggcac aagctggagt acaacctgcc ggaccaactg actgaagagc
     4981 agatcgcaga atttaaagag gaattctccc tatttgacaa ggacggggat gggacaataa
     5041 caaccaagga gctggggacg gtgatgcggt ctctggggca gaaccccaca gaagcagagc
     5101 tgcaggacat gatcaatgaa gtagatgccg acggtgacgg cacaatcgac ttccctgagt
     5161 tcctgacaat gatggcaaga aaaatgaaat acagggacac ggaagaagaa attagagaag
     5221 cgttcggtgt gtttgataag gatggcaatg gctacatcag tgcagcagag cttcgccacg
     5281 tgatgacaaa ccttggagag aagttaacag atgaagaggt tgatgaaatg atcagggaag
     5341 cagacatcga tggggatggt caggtaaact acgaagagtt tgtacaaatg atgacagcga
     5401 agtaaggtgt acaaagataa atggcgcgtt aagtgcaaca cgtgaaggcc ggccctgcag
     5461 gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     5521 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     5581 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     5641 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     5701 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     5761 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     5821 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     5881 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     5941 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     6001 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     6061 gccgcctccc cgcatcgata ccgtcgacct cgactgtgcc ttctagttgc cagccatctg
     6121 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     6181 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     6241 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     6301 aactagtggt gccagggcgt gcccttgggc tccccgggcg cggcggccgc atcgaattct
     6361 accgggtagg ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc
     6421 tgggcacttg gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta
     6481 ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag
     6541 tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag
     6601 cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc
     6661 tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa
     6721 ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt
     6781 cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct
     6841 cttcctcatc tccgggcctt tcgacctgca gcagcacgtg ttgacaatta atcatcggca
     6901 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccgctag caccatggga
     6961 tcggccattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     7021 ttcggctatg actgggcaca acagacgatc ggctgctctg atgccgccgt gttccggctg
     7081 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     7141 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     7201 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     7261 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     7321 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     7381 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     7441 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     7501 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     7561 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     7621 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     7681 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     7741 cttgacgagt tcttctgagg ggatccgctg taagtctgca gaaattgatg atctattaaa
     7801 caataaagat gtccactaaa atggaagttt ttcctgtcat actttgttaa gaagggtgag
     7861 aacagagtac ctacattttg aatggaagga ttggagctac gggggtgggg gtggggtggg
     7921 attagataaa tgcctgctct ttactgaagg ctctttacta ttgctttatg ataatgtttc
     7981 atagttggat atcataattt aaacaagcaa aaccaaatta agggccagct cattcctccc
     8041 actcatgatc tatagatcta tagatctctc gtgggatcat tgtttttctc ttgattccca
     8101 ctttgtggtt ctaagtactg tggtttccaa atgtgtcagt ttcatagcct gaagaacgag
     8161 atcagcagcc tctgttccac atacacttca ttctcagtat tgttttgcca agttctaatt
     8221 ccatcagaaa gcttgcagaa gatctcccca actggggtaa cctttgagtt ctctcagttg
     8281 ggggggcgcg ccctagaaga tgggcgggag tcttctgggc aggcttaaag gctaacctgg
     8341 tgtgtgggcg ttgtcctgca ggggaattga acaggtgtaa aattggaggg acaagacttc
     8401 ccacagattt tcggttttgt cgggaagttt tttaataggg gcaaatagga aaatggagga
     8461 taggagtcat ctggggttta tgcagcaaaa ctacaggtat attgcttgta tccgcctcgg
     8521 agatttccat gaggagataa agacatgtca cccgagttta tactctcctg cttagatcct
     8581 actacagtat gaaatacagt gtygcgaggt agactatgta agcagattta atcattttaa
     8641 agagcccagt acttcatatc catttctccc gctccttctg cagccttatc aaaaggtatt
     8701 tagaacactc attttagccc cattttcatt tattatactg gcttatccaa cccctagaca
     8761 gagcattggc attttccctt tcctgatctt agaagtctga tgactcatga aaccagacag
     8821 attagttaca tacaccacaa atcgaggctg tagctggggc ctcaacactg cagttctttt
     8881 ataactcctt agtacacttt ttgttgatcc tttgccttga tccttaattt tcagtgtcta
     8941 tcacctctcc cgtcaggtgg tgttccacat ttgggcctat tctcagtcca gggagtttta
     9001 caacaataga tgtattgaga atccaaccta aagcttaact ttccactccc atgaatgcct
     9061 ctctcctttt tctccattat aactgagcta twaccattaa tggtttcagg tggatgtctc
     9121 ctcccccaat atacctgatg tatctacata ttgccaggct gatattttaa gacatwaaag
     9181 gtatatttca ttattgagcc acatggtatt gattactgct actaaaattt tgtcattgta
     9241 cacatctgta aaaggtggtt ccttttggaa tgcaaagttc aggtgtttgt tgtctttcct
     9301 gacctaaggt cttgtgagct tgtatttttt ctatttaagc agtgctttct cttggactgg
     9361 cttgactcat ggcattctac acgttattgc tggtctaaat gtgattttgc caagcttctt
     9421 caggacctat aattttgctt gacttgtagc caaacacaag taaaatgatt aagcaacaaa
     9481 tgtatttgtg aagcttggtt tttaggttgt tgtgttgtgt gtgcttgtgc tctataataa
     9541 tactatccag gggctggaga ggtggctcgg agttcaagag cacagactgc tcttccagaa
     9601 gtcctgagtt caattcccag caaccacatg gtggctcaca accatctgta atgggatctg
     9661 atgccctctt ctggtgtgtc tgaagaccac aagtgtattc acattaaata aataatcctc
     9721 cttcttcttc tttttttttt tttaaagaga atwctgtctc cagtagaatt actgaagtaa
     9781 tgaaatactt tgtgtttgtt ccaatatggw agccaataat caaatactct twagcactgg
     9841 aaatgtacca aggaactatt ttatttaagt gwactgtgga cagaggagcc ataactgcag
     9901 acttgtggga tacagaagac caatgcagac ttaatgtctt ttctcttaca ctaagcaata
     9961 aagaaataaa aattgaactt ctagtatcct atttgttaaa ctgctagctt tactaacttt
    10021 tgtgcttcat ctatacaaag ctgaaagcta agtctgcagc cattactaaa catgaaagca
    10081 agtaatgata attttggatt tcaaaaatgt agggccagag tttagccagc cagtggtggt
    10141 gcttgccttt atgccttaat cccagcactc tggaggcaga gacaggcaga tctctgagtt
    10201 tgagcccagc ctggtctaca catcaagttc tatctaggat agccaggaat acacacagaa
    10261 accctgttgg ggaggggggc tctgagattt cataaaatta taattgaagc attccctaat
    10321 gagccactat ggatgtggct aaatccgtct acctttctga tgagatttgg gtattatttt
    10381 ttctgtctct gctgttggtt gggtcttttg acactgtggg ctttcttaaa gcctccttcc
    10441 ctgccatgtg gtctcttgtt tgctactaac ttcccatggc ttaaatggca tggctttttg
    10501 ccttctaagg gcagctgctg agwtttgcag cctgatttcc agggtggggt tgggaaatct
    10561 ttcaaacact aaaattgtcc tttaattttt ttttaaaaaa tgggttatat aataaacctc
    10621 ataaaatagt tatgaggagt gaggtggact aatattaatg agtccctccc ctataaaaga
    10681 gctattaagg ctttttgtct tatactaact ttttttttaa atgtggtatc tttagaacca
    10741 agggtcttag agttttagta tacagaaact gttgcatcgc ttaatcagat tttctagttt
    10801 caaatccaga gaatccaaat tcttcacagc caaagtcaaa ttaagaattt ctgactttaa
    10861 tgttatttgc tactgtgaat ataaaatgat agcttttcct gaggcagggt ctcactatgt
    10921 atctctgcct gatctgcaac aagatatgta gactaaagtt ctgcctgctt ttgtctcctg
    10981 aatactaagg ttaaaatgta gtaatacttt tggaacttgc aggtcagatt cttttatagg
    11041 ggacacacta agggagcttg ggtgatagtt ggtaaatgtg tttaagtgat gaaaacttga
    11101 attattatca ccgcaaccta ctttttaaaa aaaaaagcca ggcctgttag agcatgctaa
    11161 gggatcccta ggacttgctg agcacacaag agtagtactt ggcaggctcc tggtgagagc
    11221 atatttcaaa aaacaaggca gacaaccaag aaactacagt aaggttacct gtctttaacc
    11281 atctgcatat acacagggat attaaaatat tccaaataat atttcattca agttttcccc
    11341 catcaaattg ggacatggat ttctccggtg aataggcaga gttggaaact aaacaaatgt
    11401 tggttttgtg atttgtgaaa ttgttttcaa gtgatagtta aagcccatga gatacagaac
    11461 aaagctgcta tttcgaggtc tcttggttat actcagaagc acttctttgg gtttccctgc
    11521 actatcctga tcatgtgcta ggcctwcctt aggctgattg ttgttcaaat aacttaagtt
    11581 tcctgtcagg tgatgtcata tgatttcata tatcaaggca aaacatgtta tatatgttaa
    11641 acatttgkac ttaatgtgaa agttaggtct ttgtgggttt tgattttaat ttcaaaacct
    11701 gagctaaata agtcatttta catgtcttac atttggtgaa ttgtatattg tggtttgcag
    11761 gcaagactct ctgacctagt aaccctccta tagagcactt tgctgggtca caagtctagg
    11821 agtcaagcat ttcaccttga agttgagacg ttttgttagt gtatactagt tatatgttgg
    11881 aggacatgtt tatccagaag atattcagga ctatttttga ctgggctaag gaattgattc
    11941 tgattagcac tgttagtgag cattgagtgg cctttaggct tgaattggag tcacttgtat
    12001 atctcaaata atgctggcct tttttwaaaa gcccttgttc tttatcaccc tgttttctac
    12061 ataatttttg ttcaaagaaa tacttgtttg gatctccttt tgacaacaat agcatgtttt
    12121 caagccatat tttttttcct tttttttttt ttttttggtt tttcgagaca gggtttctct
    12181 gtatagccct ggctgtcctg gaactcactt tgtagaccag gctggcctcg aactcagaaa
    12241 tccgcctgcc tctgcctcct gagtgccggg attaaaggcg tgcaccacca cgcctggcta
    12301 agttggatat tttgtatata actataacca atactaactc cactgggtgg atttttaatt
    12361 cagtcagtag tcttaagtgg tctttattgg cccttattaa aatctactgt tcactctaac
    12421 agaggctgtt ggactagtgg sactaagcaa cttcctacgg atatactagc agataagggt
    12481 cagggataga aactagtcta gcgttttgta tacctaccag cttatactac cttgttctga
    12541 tagaaatatt taggacatct agcttatcga tccgtcgagg gggggcccgg tacccagctt
    12601 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
    12661 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
    12721 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
    12781 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
    12841 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
    12901 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
    12961 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
    13021 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
    13081 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
    13141 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
    13201 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
    13261 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
    13321 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
    13381 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
    13441 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
    13501 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
    13561 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
    13621 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
    13681 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
    13741 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
    13801 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
    13861 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
    13921 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
    13981 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
    14041 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
    14101 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
    14161 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
    14221 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
    14281 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
    14341 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
    14401 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
    14461 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
    14521 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
    14581 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
    14641 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
    14701 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
    14761 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
    14821 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
    14881 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
    14941 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
    15001 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
    15061 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
    15121 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
    15181 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
    15241 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
    15301 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
    15361 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg
    15421 taatacgact cactataggg cgaattggag ctcca
//
