LOCUS       sequence-377719-       17177 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Target a Cre-dependent GCaMP6s cassette and a tTA2 cassette into the
            mouse TIGRE locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17177)
  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 17177)
  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: Ai162 (TIT2L-GC6s-ICL-tTA2) targeting vector
FEATURES             Location/Qualifiers
     source          1..17177
                     /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      4732..4741
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4738..4746,4747..4764,4765..4767,4768..4800,4801..4803,
                     4804..4827,4828..4854,4855..4911,4912..4917,4918..5187,
                     5188..5205,5206..5640,5641..5646,5647..6090)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      4834..4843
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     5124..5145
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5251..5272)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5512..5531)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    6115..6703
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(6586..6597)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6734..6941
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        9465..9768
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9770..10047
                     /label=chicken beta-actin promoter
     intron          10048..11064
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     10988..11010
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     11072..11091
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    11152..11185
                     /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    11223..11271
                     /note="synthetic polyadenylation signal"
     polyA_signal    11280..11756
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(11791..11810)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    11835..11883
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    12062..12095
                     /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(12120..12737,12738..12743,12744..12779,12780..12782,
                     12783..12818,12819..12821,12822..12857,12858..12866)
                     /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    12909..13497
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(13380..13391)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    13528..13735
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    13749..13782
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        13809..14308
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(13844..13867)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     14241..14260
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     complement(14956..14972)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14956..14972)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14969..14991)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14980..14996
                     /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(15004..15010,15011..15028,15029..15034))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    15049..15070
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(15187..15204)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(15358..15946)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(15438..15457)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(16117..16908,16909..16977))
                     /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     16740..16759
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(16978..17082)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     17150..17168
                     /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 gcagagggtt 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 tcacgcgtgc cgccaccatg
     4741 ggttctcatc atcatcatca tcatggtatg gctagcatga ctggtggaca gcaaatgggt
     4801 cgggatctgt acgacgatga cgataaggat ctcgccacca tggtcgactc atcacgtcgt
     4861 aagtggaata agacaggtca cgcagtcaga gctataggtc ggctgagctc actcgagaac
     4921 gtctatatca aggccgacaa gcagaagaac ggcatcaagg cgaacttcca catccgccac
     4981 aacatcgagg acggcggcgt gcagctcgcc taccactacc agcagaacac ccccatcggc
     5041 gacggccccg tgctgctgcc cgacaaccac tacctgagcg tgcagtccaa actttcgaaa
     5101 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     5161 actctcggca tggacgagct gtacaagggc ggtaccggag ggagcatggt gagcaagggc
     5221 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     5281 cacaagttca gcgtgtccgg cgagggtgag ggcgatgcca cctacggcaa gctgaccctg
     5341 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     5401 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     5461 aagtccgcca tgcccgaagg ctacatccag gagcgcacca tcttcttcaa ggacgacggc
     5521 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     5581 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     5641 ctgccggacc aactgactga agagcagatc gcagaattta aagaggcttt ctccctattt
     5701 gacaaggacg gggatgggac aataacaacc aaggagctgg ggacggtgat gcggtctctg
     5761 gggcagaacc ccacagaagc agagctgcag gacatgatca atgaagtaga tgccgacggt
     5821 gacggcacaa tcgacttccc tgagttcctg acaatgatgg caagaaaaat gaaatacagg
     5881 gacacggaag aagaaattag agaagcgttc ggtgtgtttg ataaggatgg caatggctac
     5941 atcagtgcag cagagcttcg ccacgtgatg acaaaccttg gagagaagtt aacagatgaa
     6001 gaggttgatg aaatgatcag ggaagcagac atcgatgggg atggtcaggt aaactacgaa
     6061 gagtttgtac aaatgatgac agcgaagtaa acgcgttacg taaagcttgt cgataatcaa
     6121 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     6181 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     6241 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     6301 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     6361 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     6421 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     6481 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat
     6541 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     6601 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     6661 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgaca ccgagcgctg
     6721 ctcgaggcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     6781 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     6841 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     6901 aagggggagg attgggaaga caatagcagg catgctgggg acacgtggga tcctctagag
     6961 ccccatcctc actgactccg tcctggagtt ggatgagaga taatggcctt acgttgtgcc
     7021 aggggagggt cgggctggat ttagcaagat ttaccttctc caaagagcgg tgctgcagtg
     7081 gcacagctgc ccacggaggt gggggggtca ccgtccctgg aggtgatgaa gaactgtggg
     7141 gatgtggcac tgagggacat ggccagtggg cacggtgggt gggttggggt tggtcttggg
     7201 gatcttggag ggcttttcca gccttcatga tttgacgatt gtatgaacat ctacatggca
     7261 attctccagc tgcctgtccc agtcctactg acccagctgt atctctccag gcaagctctt
     7321 ccaccccttc tgcttgcatc cagacaccat caaacatgca ggctcagaca cagggaccag
     7381 cagtgtctgt ggcctttttg tgctcctctc catgctgggt tttaacttgc tctttgtcct
     7441 tctatcctat cttcttatcc ttaaggctgt tctgaacgct gtgacttgga gagtgtccca
     7501 gagccctcaa cacctgcatg tcccacgtcc atgctgtcct gcacttcctt atccccaaga
     7561 tctgcctctc cgtgatgcac tgaattggca aacatgtgtc accccagacc aacaatgtca
     7621 cagcaaactc ccccttgata ggacaagggg gaatggcttt acactgagac aggggaggtt
     7681 tgggttggat atgaggaggc agtttttccc ccagagggtg gtgacgcact gaacaggttg
     7741 cccaaggagg ctgtggatgc cccatccctg caggcattca aggccaggct ggatgtggct
     7801 ctgggcagcc tgggctgctg gttgatgacc ctgcacatag cagggggttg gatctggatg
     7861 agcactgtgc tcctttgcaa cccaggccgt tctatgattc tgtcattcta aatctctctt
     7921 tcagcctaaa gctttttccc cgtatccccc caggtgtctg caggctcaaa gagcagcgag
     7981 aagcgttcag aggaaagcga tcccgtgcca ccttccccgt gcccgggctg tccccgcacg
     8041 ctgccggctc ggggatgcgg ggggagcgcc ggaccggagc ggagccccgg gcggctcgct
     8101 gctgccccct agcgggggag ggacgtaatt acatccctgg gggctttggg ggggggctgt
     8161 ccctctagag ccccatcctc actgactccg tcctggagtt ggatgagaga taatggcctt
     8221 acgttgtgcc aggggagggt cgggctggat ttagcaagat ttaccttctc caaagagcgg
     8281 tgctgcagtg gcacagctgc ccacggaggt gggggggtca ccgtccctgg aggtgatgaa
     8341 gaactgtggg gatgtggcac tgagggacat ggccagtggg cacggtgggt gggttggggt
     8401 tggtcttggg gatcttggag ggcttttcca gccttcatga tttgacgatt gtatgaacat
     8461 ctacatggca attctccagc tgcctgtccc agtcctactg acccagctgt atctctccag
     8521 gcaagctctt ccaccccttc tgcttgcatc cagacaccat caaacatgca ggctcagaca
     8581 cagggaccag cagtgtctgt ggcctttttg tgctcctctc catgctgggt tttaacttgc
     8641 tctttgtcct tctatcctat cttcttatcc ttaaggctgt tctgaacgct gtgacttgga
     8701 gagtgtccca gagccctcaa cacctgcatg tcccacgtcc atgctgtcct gcacttcctt
     8761 atccccaaga tctgcctctc cgtgatgcac tgaattggca aacatgtgtc accccagacc
     8821 aacaatgtca cagcaaactc ccccttgata ggacaagggg gaatggcttt acactgagac
     8881 aggggaggtt tgggttggat atgaggaggc agtttttccc ccagagggtg gtgacgcact
     8941 gaacaggttg cccaaggagg ctgtggatgc cccatccctg caggcattca aggccaggct
     9001 ggatgtggct ctgggcagcc tgggctgctg gttgatgacc ctgcacatag cagggggttg
     9061 gatctggatg agcactgtgc tcctttgcaa cccaggccgt tctatgattc tgtcattcta
     9121 aatctctctt tcagcctaaa gctttttccc cgtatccccc caggtgtctg caggctcaaa
     9181 gagcagcgag aagcgttcag aggaaagcga tcccgtgcca ccttccccgt gcccgggctg
     9241 tccccgcacg ctgccggctc ggggatgcgg ggggagcgcc ggaccggagc ggagccccgg
     9301 gcggctcgct gctgccccct agcgggggag ggacgtaatt acatccctgg gggctttggg
     9361 ggggggctgt ccctctagag tcgacggcgc gccctgcagg aatctagtta ttaatagtaa
     9421 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     9481 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     9541 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     9601 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     9661 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     9721 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc
     9781 cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta
     9841 tttatttttt aattattttg tgcagcgatg ggggcggggg gggggggggg gcgcgcgccr
     9901 ggsggggsgg ggsggggsgr ggggsggggs ggggsgaggc ggagaggtgc ggcggcagcc
     9961 aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc
    10021 tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc
    10081 cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag
    10141 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg
    10201 cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc
    10261 ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc
    10321 tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc
    10381 agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag
    10441 gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg
    10501 tcggtcgggc tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct
    10561 tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg
    10621 cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag
    10681 gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc
    10741 cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag
    10801 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc
    10861 gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt
    10921 ctccctctcc agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc
    10981 agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt
    11041 catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat
    11101 cattttggca aagaattgat ttgataccgc gggccccgcg tggacggtac cataacttcg
    11161 tataaagtat cctatacgaa gttatccatg gctagcggca gcctcggagt ttgaatagat
    11221 agaataaaat atctttattt tcattccatc tgtgtgttgg ttttttgtgt gagatctacg
    11281 ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag
    11341 tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct
    11401 tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa
    11461 cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc
    11521 tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcaggct cccgagttgt
    11581 tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg
    11641 gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt
    11701 ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctgc
    11761 ctcagctaat tgagtagggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
    11821 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg
    11881 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
    11941 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
    12001 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccg tggaagttcc tattctctag
    12061 aataacttcg tataaagtat cctatacgaa gttatacgcg cgcgtccatt taattcacca
    12121 tgtctagact ggacaagagc aaagtcataa actctgctct ggaattactc aatgaagtcg
    12181 gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct gggagttgag cagcctaccc
    12241 tgtactggca cgtgaagaac aagcgggccc tgctcgatgc cctggcaatc gagatgctgg
    12301 acaggcatca tacccacttc tgccccctgg aaggcgagtc atggcaagac tttctgcgga
    12361 acaacgccaa gtcattccgc tgtgctctcc tctcacatcg cgacggggct aaagtgcatc
    12421 tcggcacccg cccaacagag aaacagtacg aaaccctgga aaatcagctc gcgttcctgt
    12481 gtcagcaagg cttctccctg gagaacgcac tgtacgctct gtccgccgtg ggccacttta
    12541 cactgggctg cgtattggag gatcaggagc atcaagtagc aaaagaggaa agagagacac
    12601 ctaccaccga ttctatgccc ccacttctga gacaagcaat tgagctgttc gaccatcagg
    12661 gagccgaacc tgccttcctt ttcggcctgg aactaatcat atgtggcctg gagaaacagc
    12721 taaagtgcga aagcggcggg ccggccgacg cccttgacga ttttgactta gacatgctcc
    12781 cagccgatgc ccttgacgac tttgaccttg atatgctgcc tgctgacgct cttgacgatt
    12841 ttgaccttga catgctcccc gggtaactaa gtaaggatca aatggcgcgt tacgtaaagc
    12901 ttgtcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
    12961 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
    13021 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
    13081 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
    13141 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
    13201 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
    13261 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
    13321 ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
    13381 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
    13441 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
    13501 gacaccgagc gctgctcgag gcctcgactg tgccttctag ttgccagcca tctgttgttt
    13561 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
    13621 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
    13681 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggacacgt
    13741 gggatccggt gccagggcgt gcccttgggc tccccgggcg cgtttgggcg gccgcatcga
    13801 attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc
    13861 cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac
    13921 cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc
    13981 cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga
    14041 agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt
    14101 aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct
    14161 gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg
    14221 aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt
    14281 ctcctcttcc tcatctccgg gcctttcgac ctgcaatcgc cgctagccgc caccatgaaa
    14341 aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc
    14401 tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga
    14461 gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat
    14521 gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa
    14581 ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac
    14641 ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc
    14701 gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt
    14761 caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg
    14821 caaactgtga tggacgacac cgtcagtgta agtgctagcc cctaggcctg aatcgaagtt
    14881 cctattcttc aaaaggtata ggaacttcga gctcgtcgac tgcagaggcc tgcatgcaag
    14941 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
    15001 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
    15061 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
    15121 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
    15181 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
    15241 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
    15301 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
    15361 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
    15421 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
    15481 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
    15541 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
    15601 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
    15661 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
    15721 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
    15781 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
    15841 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
    15901 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
    15961 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
    16021 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
    16081 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
    16141 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
    16201 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
    16261 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
    16321 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
    16381 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
    16441 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
    16501 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
    16561 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
    16621 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
    16681 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
    16741 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
    16801 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
    16861 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
    16921 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
    16981 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
    17041 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
    17101 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
    17161 cacgaggccc tttcgtc
//
