LOCUS       sequence-227312-       17206 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Target a Cre-dependent GCaMP6f expression 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 17206)
  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 17206)
  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: Ai148 (TIT2L-GC6f-ICL-tTA2) targeting vector
FEATURES             Location/Qualifiers
     source          1..17206
                     /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    2975..3056
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3000..3019
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    join(3070..3088,3089..3111,3112..3130,3131..3153,3154..3172,
                     3173..3195,3196..3214,3215..3237,3238..3256,3257..3279,
                     3280..3298,3299..3321,3322..3340)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        3373..3411
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3408..3432
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    complement(3545..3578)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3655..3703
                     /note="synthetic polyadenylation signal"
     polyA_signal    3712..4188
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     polyA_signal    4200..4661
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     protein_bind    complement(4688..4721)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      4747..4756
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4753..4761,4762..4779,4780..4782,4783..4815,4816..4818,
                     4819..4842,4843..4869,4870..4926,4927..4932,4933..5202,
                     5203..5220,5221..5655,5656..5661,5662..6105)
                     /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      4849..4858
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     5139..5160
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5266..5287)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5527..5546)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    6148..6736
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(6619..6630)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6767..6974
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        9498..9801
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9803..10080
                     /label=chicken beta-actin promoter
     intron          10081..11097
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     11021..11043
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     11105..11124
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    11185..11218
                     /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    11256..11304
                     /note="synthetic polyadenylation signal"
     polyA_signal    11313..11789
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(11824..11843)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    11868..11916
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    12095..12128
                     /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(12149..12766,12767..12772,12773..12808,12809..12811,
                     12812..12847,12848..12850,12851..12886,12887..12895)
                     /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    12938..13526
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(13409..13420)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    13557..13764
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    13778..13811
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        13838..14337
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(13873..13896)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     14270..14289
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     complement(14985..15001)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14985..15001)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14998..15020)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    15009..15025
                     /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(15033..15039,15040..15057,15058..15063))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    15078..15099
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(15216..15233)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(15387..15975)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(15467..15486)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(16146..16937,16938..17006))
                     /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     16769..16788
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(17007..17111)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     17179..17197
                     /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 gctgtccctc tagcgagctc ggtacccaga gccccatcct cactgactcc
     1741 gtcctggagt tggatgagag ataatggcct tacgttgtgc caggggaggg tcgggctgga
     1801 tttagcaaga tttaccttct ccaaagagcg gtgctgcagt ggcacagctg cccacggagg
     1861 tgggggggtc accgtccctg gaggtgatga agaactgtgg ggatgtggca ctgagggaca
     1921 tggccagtgg gcacggtggg tgggttgggg ttggtcttgg ggatcttgga gggcttttcc
     1981 agccttcatg atttgacgat tgtatgaaca tctacatggc aattctccag ctgcctgtcc
     2041 cagtcctact gacccagctg tatctctcca ggcaagctct tccacccctt ctgcttgcat
     2101 ccagacacca tcaaacatgc aggctcagac acagggacca gcagtgtctg tggccttttt
     2161 gtgctcctct ccatgctggg ttttaacttg ctctttgtcc ttctatccta tcttcttatc
     2221 cttaaggctg ttctgaacgc tgtgacttgg agagtgtccc agagccctca acacctgcat
     2281 gtcccacgtc catgctgtcc tgcacttcct tatccccaag atctgcctct ccgtgatgca
     2341 ctgaattggc aaacatgtgt caccccagac caacaatgtc acagcaaact cccccttgat
     2401 aggacaaggg ggaatggctt tacactgaga caggggaggt ttgggttgga tatgaggagg
     2461 cagtttttcc cccagagggt ggtgacgcac tgaacaggtt gcccaaggag gctgtggatg
     2521 ccccatccct gcaggcattc aaggccaggc tggatgtggc tctgggcagc ctgggctgct
     2581 ggttgatgac cctgcacata gcagggggtt ggatctggat gagcactgtg ctcctttgca
     2641 acccaggccg ttctatgatt ctgtcattct aaatctctct ttcagcctaa agctttttcc
     2701 ccgtatcccc ccaggtgtct gcaggctcaa agagcagcga gaagcgttca gaggaaagcg
     2761 atcccgtgcc accttccccg tgcccgggct gtccccgcac gctgccggct cggggatgcg
     2821 gggggagcgc cggaccggag cggagccccg ggcggctcgc tgctgccccc tagcggggga
     2881 gggacgtaat tacatccctg ggggctttgg gggggggctg tccctctagg ggtacccggg
     2941 gatccaccgg tatcgattcg gccggccgac gtcggcagtg aaaaaaatgc tttatttgtg
     3001 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagtttcga
     3061 gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca
     3121 gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa
     3181 agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc
     3241 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
     3301 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagctcgg
     3361 tacccgggtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg
     3421 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
     3481 gaccgatcca gcctccgcgg ccccgaattc gagctcggta cccggggatc aaattcaatt
     3541 aaatataact tcgtatagca tacattatac gaagttattc gcgatgaata aatgaaagct
     3601 tgcagatccc ctcagcacca tggctagcgg cagcctcgga gtttgaatag atagaataaa
     3661 atatctttat tttcattcca tctgtgtgtt ggttttttgt gtgagatcta cgggtggcat
     3721 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     3781 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     3841 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     3901 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     3961 atctccgcct cctgggttca agcgattctc ctgcctcagg ctcccgagtt gttgggattc
     4021 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     4081 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc
     4141 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gcctcagcgc
     4201 tgtaagtctg cagaaattga tgatctatta aacaataaag atgtccacta aaatggaagt
     4261 ttttcctgtc atactttgtt aagaagggtg agaacagagt acctacattt tgaatggaag
     4321 gattggagct acgggggtgg gggtggggtg ggattagata aatgcctgct ctttactgaa
     4381 ggctctttac tattgcttta tgataatgtt tcatagttgg atatcataat ttaaacaagc
     4441 aaaaccaaat taagggccag ctcattcctc ccactcatga tctatagatc tatagatctc
     4501 tcgtgggatc attgtttttc tcttgattcc cactttgtgg ttctaagtac tgtggtttcc
     4561 aaatgtgtca gtttcatagc ctgaagaacg agatcagcag cctctgttcc acatacactt
     4621 cattctcagt attgttttgc caagttctaa ttccatcaga aagcttttaa ttaactgaat
     4681 agctaacata acttcgtata gcatacatta tacgaagtta tcacgcgcca tttgggaacc
     4741 ggtgccgcca ccatgggttc tcatcatcat catcatcatg gtatggctag catgactggt
     4801 ggacagcaaa tgggtcggga tctgtacgac gatgacgata aggatctcgc caccatggtc
     4861 gactcatcac gtcgtaagtg gaataagaca ggtcacgcag tcagagctat aggtcggctg
     4921 agctcactcg agaacgtcta tatcaaggcc gacaagcaga agaacggcat caaggcgaac
     4981 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgcctacca ctaccagcag
     5041 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcgtgcag
     5101 tccaaacttt cgaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     5161 accgccgccg ggatcactct cggcatggac gagctgtaca agggcggtac cggagggagc
     5221 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     5281 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gtgagggcga tgccacctac
     5341 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     5401 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     5461 cagcacgact tcttcaagtc cgccatgccc gaaggctaca tccaggagcg caccatcttc
     5521 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     5581 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     5641 aagctggagt acaacctgcc ggaccaactg actgaagagc agatcgcaga atttaaagag
     5701 gaattctccc tatttgacaa ggacggggat gggacaataa caaccaagga gctggggacg
     5761 gtgatgcggt ctctggggca gaaccccaca gaagcagagc tgcaggacat gatcaatgaa
     5821 gtagatgccg acggtgacgg cacaatcgac ttccctgagt tcctgacaat gatggcaaga
     5881 aaaatgaaat acagggacac ggaagaagaa attagagaag cgttcggtgt gtttgataag
     5941 gatggcaatg gctacatcag tgcagcagag cttcgccacg tgatgacaaa ccttggagag
     6001 aagttaacag atgaagaggt tgatgaaatg atcagggaag cagacatcga tggggatggt
     6061 caggtaaact acgaagagtt tgtacaaatg atgacagcga agtaaggtgt acaaagataa
     6121 atggcgcgtt acgtaaagct tgtcgataat caacctctgg attacaaaat ttgtgaaaga
     6181 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     6241 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     6301 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     6361 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     6421 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     6481 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     6541 aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg
     6601 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     6661 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     6721 tgggccgcct ccccgcatcg acaccgagcg ctgctcgagg cctcgactgt gccttctagt
     6781 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     6841 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     6901 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     6961 aggcatgctg gggacacgtg ggatcctcta gagccccatc ctcactgact ccgtcctgga
     7021 gttggatgag agataatggc cttacgttgt gccaggggag ggtcgggctg gatttagcaa
     7081 gatttacctt ctccaaagag cggtgctgca gtggcacagc tgcccacgga ggtggggggg
     7141 tcaccgtccc tggaggtgat gaagaactgt ggggatgtgg cactgaggga catggccagt
     7201 gggcacggtg ggtgggttgg ggttggtctt ggggatcttg gagggctttt ccagccttca
     7261 tgatttgacg attgtatgaa catctacatg gcaattctcc agctgcctgt cccagtccta
     7321 ctgacccagc tgtatctctc caggcaagct cttccacccc ttctgcttgc atccagacac
     7381 catcaaacat gcaggctcag acacagggac cagcagtgtc tgtggccttt ttgtgctcct
     7441 ctccatgctg ggttttaact tgctctttgt ccttctatcc tatcttctta tccttaaggc
     7501 tgttctgaac gctgtgactt ggagagtgtc ccagagccct caacacctgc atgtcccacg
     7561 tccatgctgt cctgcacttc cttatcccca agatctgcct ctccgtgatg cactgaattg
     7621 gcaaacatgt gtcaccccag accaacaatg tcacagcaaa ctcccccttg ataggacaag
     7681 ggggaatggc tttacactga gacaggggag gtttgggttg gatatgagga ggcagttttt
     7741 cccccagagg gtggtgacgc actgaacagg ttgcccaagg aggctgtgga tgccccatcc
     7801 ctgcaggcat tcaaggccag gctggatgtg gctctgggca gcctgggctg ctggttgatg
     7861 accctgcaca tagcaggggg ttggatctgg atgagcactg tgctcctttg caacccaggc
     7921 cgttctatga ttctgtcatt ctaaatctct ctttcagcct aaagcttttt ccccgtatcc
     7981 ccccaggtgt ctgcaggctc aaagagcagc gagaagcgtt cagaggaaag cgatcccgtg
     8041 ccaccttccc cgtgcccggg ctgtccccgc acgctgccgg ctcggggatg cggggggagc
     8101 gccggaccgg agcggagccc cgggcggctc gctgctgccc cctagcgggg gagggacgta
     8161 attacatccc tgggggcttt gggggggggc tgtccctcta gagccccatc ctcactgact
     8221 ccgtcctgga gttggatgag agataatggc cttacgttgt gccaggggag ggtcgggctg
     8281 gatttagcaa gatttacctt ctccaaagag cggtgctgca gtggcacagc tgcccacgga
     8341 ggtggggggg tcaccgtccc tggaggtgat gaagaactgt ggggatgtgg cactgaggga
     8401 catggccagt gggcacggtg ggtgggttgg ggttggtctt ggggatcttg gagggctttt
     8461 ccagccttca tgatttgacg attgtatgaa catctacatg gcaattctcc agctgcctgt
     8521 cccagtccta ctgacccagc tgtatctctc caggcaagct cttccacccc ttctgcttgc
     8581 atccagacac catcaaacat gcaggctcag acacagggac cagcagtgtc tgtggccttt
     8641 ttgtgctcct ctccatgctg ggttttaact tgctctttgt ccttctatcc tatcttctta
     8701 tccttaaggc tgttctgaac gctgtgactt ggagagtgtc ccagagccct caacacctgc
     8761 atgtcccacg tccatgctgt cctgcacttc cttatcccca agatctgcct ctccgtgatg
     8821 cactgaattg gcaaacatgt gtcaccccag accaacaatg tcacagcaaa ctcccccttg
     8881 ataggacaag ggggaatggc tttacactga gacaggggag gtttgggttg gatatgagga
     8941 ggcagttttt cccccagagg gtggtgacgc actgaacagg ttgcccaagg aggctgtgga
     9001 tgccccatcc ctgcaggcat tcaaggccag gctggatgtg gctctgggca gcctgggctg
     9061 ctggttgatg accctgcaca tagcaggggg ttggatctgg atgagcactg tgctcctttg
     9121 caacccaggc cgttctatga ttctgtcatt ctaaatctct ctttcagcct aaagcttttt
     9181 ccccgtatcc ccccaggtgt ctgcaggctc aaagagcagc gagaagcgtt cagaggaaag
     9241 cgatcccgtg ccaccttccc cgtgcccggg ctgtccccgc acgctgccgg ctcggggatg
     9301 cggggggagc gccggaccgg agcggagccc cgggcggctc gctgctgccc cctagcgggg
     9361 gagggacgta attacatccc tgggggcttt gggggggggc tgtccctcta gagtcgacgg
     9421 cgcgccctgc aggaatctag ttattaatag taatcaatta cggggtcatt agttcatagc
     9481 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     9541 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     9601 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     9661 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     9721 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     9781 ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc
     9841 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
     9901 atgggggcgg gggggggggg ggggcgcgcg ccrggsgggg sggggsgggg sgrggggsgg
     9961 ggsggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
    10021 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
    10081 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
    10141 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
    10201 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
    10261 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
    10321 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
    10381 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
    10441 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
    10501 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
    10561 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
    10621 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
    10681 gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc
    10741 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
    10801 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
    10861 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
    10921 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
    10981 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
    11041 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
    11101 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt gatttgatac
    11161 cgcgggcccc gcgtggacgg taccataact tcgtataaag tatcctatac gaagttatcc
    11221 atggctagcg gcagcctcgg agtttgaata gatagaataa aatatcttta ttttcattcc
    11281 atctgtgtgt tggttttttg tgtgagatct acgggtggca tccctgtgac ccctccccag
    11341 tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat
    11401 taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg
    11461 tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg
    11521 aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc
    11581 aagcgattct cctgcctcag gctcccgagt tgttgggatt ccaggcatgc atgaccaggc
    11641 tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct
    11701 ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg
    11761 cgtgaaccac tgctcccttc cctgtccttc tgcctcagct aattgagtag gggggaggct
    11821 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
    11881 acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
    11941 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
    12001 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
    12061 gggcggcagg ccgtggaagt tcctattctc tagaataact tcgtataaag tatcctatac
    12121 gaagttatac gcgtccattt aattcaccat gtctagactg gacaagagca aagtcataaa
    12181 ctctgctctg gaattactca atgaagtcgg tatcgaaggc ctgacgacaa ggaaactcgc
    12241 tcaaaagctg ggagttgagc agcctaccct gtactggcac gtgaagaaca agcgggccct
    12301 gctcgatgcc ctggcaatcg agatgctgga caggcatcat acccacttct gccccctgga
    12361 aggcgagtca tggcaagact ttctgcggaa caacgccaag tcattccgct gtgctctcct
    12421 ctcacatcgc gacggggcta aagtgcatct cggcacccgc ccaacagaga aacagtacga
    12481 aaccctggaa aatcagctcg cgttcctgtg tcagcaaggc ttctccctgg agaacgcact
    12541 gtacgctctg tccgccgtgg gccactttac actgggctgc gtattggagg atcaggagca
    12601 tcaagtagca aaagaggaaa gagagacacc taccaccgat tctatgcccc cacttctgag
    12661 acaagcaatt gagctgttcg accatcaggg agccgaacct gccttccttt tcggcctgga
    12721 actaatcata tgtggcctgg agaaacagct aaagtgcgaa agcggcgggc cggccgacgc
    12781 ccttgacgat tttgacttag acatgctccc agccgatgcc cttgacgact ttgaccttga
    12841 tatgctgcct gctgacgctc ttgacgattt tgaccttgac atgctccccg ggtaactaag
    12901 taaggatcaa atggcgcgtt acgtaaagct tgtcgataat caacctctgg attacaaaat
    12961 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
    13021 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
    13081 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
    13141 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
    13201 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
    13261 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
    13321 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca cctggattct
    13381 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
    13441 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
    13501 gatctccctt tgggccgcct ccccgcatcg acaccgagcg ctgctcgagg cctcgactgt
    13561 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
    13621 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
    13681 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
    13741 agacaatagc aggcatgctg gggacacgtg ggatccggtg ccagggcgtg cccttgggct
    13801 ccccgggcgc gtttgggcgg ccgcatcgaa ttctaccggg taggggaggc gcttttccca
    13861 aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc
    13921 ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt
    13981 ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag
    14041 ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg
    14101 gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt
    14161 tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg
    14221 ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg
    14281 cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgacc
    14341 tgcaatcgcc gctagccgcc accatgaaaa agcctgaact caccgcgacg tctgtcgaga
    14401 agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag
    14461 aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct
    14521 gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc
    14581 cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc
    14641 gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc
    14701 agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt
    14761 tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg
    14821 cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgtaa
    14881 gtgctagccc ctaggcctga atcgaagttc ctattcttca aaaggtatag gaacttcgag
    14941 ctcgtcgact gcagaggcct gcatgcaagc ttggcgtaat catggtcata gctgtttcct
    15001 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
    15061 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
    15121 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
    15181 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
    15241 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
    15301 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
    15361 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
    15421 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
    15481 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
    15541 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
    15601 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
    15661 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
    15721 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
    15781 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
    15841 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
    15901 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
    15961 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
    16021 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
    16081 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
    16141 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
    16201 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
    16261 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
    16321 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
    16381 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
    16441 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
    16501 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
    16561 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
    16621 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
    16681 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
    16741 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
    16801 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
    16861 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
    16921 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
    16981 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
    17041 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
    17101 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
    17161 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
