LOCUS       sequence-227286-       18030 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Target a Cre-dependent GFP and a tdTomato-P2A-tTA2 expression 
            cassette into the mouse TIGRE locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18030)
  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 18030)
  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: Ai139 (TIT2L-GFP-ICL-TPT) targeting vector
FEATURES             Location/Qualifiers
     source          1..18030
                     /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      4735..4744
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4741..4743,4744..4746,4747..5460)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4741..4743,4744..4746,4747..5457)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4786..4807)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5047..5066)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5394..5415
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5485..6073
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(5956..5967)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6104..6311
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        8835..9138
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9140..9417
                     /label=chicken beta-actin promoter
     intron          9418..10434
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     10358..10380
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     10442..10461
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    10522..10555
                     /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    10593..10641
                     /note="synthetic polyadenylation signal"
     polyA_signal    10650..11126
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(11161..11180)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    11205..11253
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    11427..11460
                     /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)."
     regulatory      11473..11482
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(11612..11630)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     11880..11899
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     12074..12097
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             12916..12972
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(12973..13590,13591..13596,13597..13632,13633..13635,
                     13636..13671,13672..13674,13675..13710,13711..13719)
                     /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    13762..14350
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(14233..14244)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    14381..14588
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    14602..14635
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        14662..15161
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(14697..14720)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     15094..15113
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     complement(15809..15825)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(15809..15825)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(15822..15844)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    15833..15849
                     /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(15857..15863,15864..15881,15882..15887))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    15902..15923
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(16040..16057)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(16211..16799)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(16291..16310)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(16970..17761,17762..17830))
                     /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     17593..17612
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(17831..17935)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     18003..18021
                     /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 tagcgccacc
     4741 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     4801 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     4861 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     4921 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     4981 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     5041 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     5101 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     5161 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     5221 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     5281 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     5341 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     5401 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     5461 acgcgttacg taaagcttgt 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 gctcgcctat gttgccacct ggattctgcg cgggacgtcc
     5941 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     6001 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     6061 gccgcctccc cgcatcgaca ccgagcgctg ctcgaggcct cgactgtgcc ttctagttgc
     6121 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     6181 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     6241 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     6301 catgctgggg acacgtggga tcctctagag ccccatcctc actgactccg tcctggagtt
     6361 ggatgagaga taatggcctt acgttgtgcc aggggagggt cgggctggat ttagcaagat
     6421 ttaccttctc caaagagcgg tgctgcagtg gcacagctgc ccacggaggt gggggggtca
     6481 ccgtccctgg aggtgatgaa gaactgtggg gatgtggcac tgagggacat ggccagtggg
     6541 cacggtgggt gggttggggt tggtcttggg gatcttggag ggcttttcca gccttcatga
     6601 tttgacgatt gtatgaacat ctacatggca attctccagc tgcctgtccc agtcctactg
     6661 acccagctgt atctctccag gcaagctctt ccaccccttc tgcttgcatc cagacaccat
     6721 caaacatgca ggctcagaca cagggaccag cagtgtctgt ggcctttttg tgctcctctc
     6781 catgctgggt tttaacttgc tctttgtcct tctatcctat cttcttatcc ttaaggctgt
     6841 tctgaacgct gtgacttgga gagtgtccca gagccctcaa cacctgcatg tcccacgtcc
     6901 atgctgtcct gcacttcctt atccccaaga tctgcctctc cgtgatgcac tgaattggca
     6961 aacatgtgtc accccagacc aacaatgtca cagcaaactc ccccttgata ggacaagggg
     7021 gaatggcttt acactgagac aggggaggtt tgggttggat atgaggaggc agtttttccc
     7081 ccagagggtg gtgacgcact gaacaggttg cccaaggagg ctgtggatgc cccatccctg
     7141 caggcattca aggccaggct ggatgtggct ctgggcagcc tgggctgctg gttgatgacc
     7201 ctgcacatag cagggggttg gatctggatg agcactgtgc tcctttgcaa cccaggccgt
     7261 tctatgattc tgtcattcta aatctctctt tcagcctaaa gctttttccc cgtatccccc
     7321 caggtgtctg caggctcaaa gagcagcgag aagcgttcag aggaaagcga tcccgtgcca
     7381 ccttccccgt gcccgggctg tccccgcacg ctgccggctc ggggatgcgg ggggagcgcc
     7441 ggaccggagc ggagccccgg gcggctcgct gctgccccct agcgggggag ggacgtaatt
     7501 acatccctgg gggctttggg ggggggctgt ccctctagag ccccatcctc actgactccg
     7561 tcctggagtt ggatgagaga taatggcctt acgttgtgcc aggggagggt cgggctggat
     7621 ttagcaagat ttaccttctc caaagagcgg tgctgcagtg gcacagctgc ccacggaggt
     7681 gggggggtca ccgtccctgg aggtgatgaa gaactgtggg gatgtggcac tgagggacat
     7741 ggccagtggg cacggtgggt gggttggggt tggtcttggg gatcttggag ggcttttcca
     7801 gccttcatga tttgacgatt gtatgaacat ctacatggca attctccagc tgcctgtccc
     7861 agtcctactg acccagctgt atctctccag gcaagctctt ccaccccttc tgcttgcatc
     7921 cagacaccat caaacatgca ggctcagaca cagggaccag cagtgtctgt ggcctttttg
     7981 tgctcctctc catgctgggt tttaacttgc tctttgtcct tctatcctat cttcttatcc
     8041 ttaaggctgt tctgaacgct gtgacttgga gagtgtccca gagccctcaa cacctgcatg
     8101 tcccacgtcc atgctgtcct gcacttcctt atccccaaga tctgcctctc cgtgatgcac
     8161 tgaattggca aacatgtgtc accccagacc aacaatgtca cagcaaactc ccccttgata
     8221 ggacaagggg gaatggcttt acactgagac aggggaggtt tgggttggat atgaggaggc
     8281 agtttttccc ccagagggtg gtgacgcact gaacaggttg cccaaggagg ctgtggatgc
     8341 cccatccctg caggcattca aggccaggct ggatgtggct ctgggcagcc tgggctgctg
     8401 gttgatgacc ctgcacatag cagggggttg gatctggatg agcactgtgc tcctttgcaa
     8461 cccaggccgt tctatgattc tgtcattcta aatctctctt tcagcctaaa gctttttccc
     8521 cgtatccccc caggtgtctg caggctcaaa gagcagcgag aagcgttcag aggaaagcga
     8581 tcccgtgcca ccttccccgt gcccgggctg tccccgcacg ctgccggctc ggggatgcgg
     8641 ggggagcgcc ggaccggagc ggagccccgg gcggctcgct gctgccccct agcgggggag
     8701 ggacgtaatt acatccctgg gggctttggg ggggggctgt ccctctagag tcgacggcgc
     8761 gccctgcagg aatctagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     8821 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     8881 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     8941 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     9001 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     9061 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     9121 gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc
     9181 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg
     9241 ggggcggggg gggggggggg gcgcgcgccr ggsggggsgg ggsggggcgr ggggsggggc
     9301 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
     9361 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
     9421 gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc
     9481 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
     9541 ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc
     9601 cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg
     9661 tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc
     9721 gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc
     9781 ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc
     9841 gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc
     9901 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc
     9961 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
    10021 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
    10081 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
    10141 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
    10201 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
    10261 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
    10321 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
    10381 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
    10441 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattgat ttgataccgc
    10501 gggccccgcg tggacggtac cataacttcg tataaagtat cctatacgaa gttatccatg
    10561 gctagcggca gcctcggagt ttgaatagat agaataaaat atctttattt tcattccatc
    10621 tgtgtgttgg ttttttgtgt gagatctacg ggtggcatcc ctgtgacccc tccccagtgc
    10681 ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa
    10741 gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg
    10801 tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac
    10861 caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag
    10921 cgattctcct gcctcaggct cccgagttgt tgggattcca ggcatgcatg accaggctca
    10981 gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca
    11041 actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt
    11101 gaaccactgc tcccttccct gtccttctgc ctcagctaat tgagtagggg ggaggctaac
    11161 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
    11221 gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
    11281 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
    11341 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
    11401 cggcaggcct cgcgagggtt tctagaataa cttcgtataa agtatcctat acgaagttat
    11461 acgcgtacta gcgccaccat ggtgagcaag ggcgaggagg tcatcaaaga gttcatgcgc
    11521 ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc
    11581 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
    11641 ctgcccttcg cctgggacat cctgtccccc cagttcatgt acggctccaa ggcgtacgtg
    11701 aagcaccccg ccgacatccc cgattacaag aagctgtcct tccccgaggg cttcaagtgg
    11761 gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg tgacccagga ctcctccctg
    11821 caggacggca cgctgatcta caaggtgaag atgcgcggca ccaacttccc ccccgacggc
    11881 cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc
    11941 gacggcgtgc tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccactac
    12001 ctggtggagt tcaagaccat ctacatggcc aagaagcccg tgcaactgcc cggctactac
    12061 tacgtggaca ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
    12121 tacgagcgct ccgagggccg ccaccacctg ttcctggggc atggtaccgg cagcaccggc
    12181 agcggcagct ccggcaccgc ctcctccgag gacaacaaca tggccgtgat taaggagttt
    12241 atgaggttta aagtcaggat ggagggcagc atgaacgggc acgagtttga gattgaagga
    12301 gagggggaag ggaggcccta tgagggcacc cagacagcaa agctgaaagt gacaaagggc
    12361 ggccctctgc cattcgcatg ggacatcctg tctcctcagt ttatgtatgg aagtaaagcc
    12421 tacgtgaaac accctgctga cattccagat tacaagaaac tgagcttccc agaggggttt
    12481 aagtgggaaa gagtgatgaa cttcgaagac gggggcctgg tgactgtcac ccaggacagc
    12541 tctctgcagg acggcactct gatctataag gtgaaaatgc gcggcaccaa ttttccccct
    12601 gatggcccag tcatgcagaa gaagacaatg ggctgggagg cttcaacaga aaggctgtac
    12661 cccagagacg gcgtgctgaa gggagagatc caccaggcac tgaagctgaa agatggaggg
    12721 cattatctgg tcgaattcaa aaccatctac atggccaaga aacccgtgca gctgcctggc
    12781 tactattacg tcgacacaaa gctggatatt actagccata acgaagacta caccattgtg
    12841 gagcagtatg agcggagcga ggggaggcac catctgtttc tgtacggcat ggacgagctg
    12901 tacaagggaa gcggagctac taacttcagc ctgctgaagc aggctggaga cgtggaggag
    12961 aaccctggac ctatgtctag actggacaag agcaaagtca taaactctgc tctggaatta
    13021 ctcaatgaag tcggtatcga aggcctgacg acaaggaaac tcgctcaaaa gctgggagtt
    13081 gagcagccta ccctgtactg gcacgtgaag aacaagcggg ccctgctcga tgccctggca
    13141 atcgagatgc tggacaggca tcatacccac ttctgccccc tggaaggcga gtcatggcaa
    13201 gactttctgc ggaacaacgc caagtcattc cgctgtgctc tcctctcaca tcgcgacggg
    13261 gctaaagtgc atctcggcac ccgcccaaca gagaaacagt acgaaaccct ggaaaatcag
    13321 ctcgcgttcc tgtgtcagca aggcttctcc ctggagaacg cactgtacgc tctgtccgcc
    13381 gtgggccact ttacactggg ctgcgtattg gaggatcagg agcatcaagt agcaaaagag
    13441 gaaagagaga cacctaccac cgattctatg cccccacttc tgagacaagc aattgagctg
    13501 ttcgaccatc agggagccga acctgccttc cttttcggcc tggaactaat catatgtggc
    13561 ctggagaaac agctaaagtg cgaaagcggc gggccggccg acgcccttga cgattttgac
    13621 ttagacatgc tcccagccga tgcccttgac gactttgacc ttgatatgct gcctgctgac
    13681 gctcttgacg attttgacct tgacatgctc cccgggtaac taagtaagga tcaaatggcg
    13741 cgttacgtaa agcttgtcga taatcaacct ctggattaca aaatttgtga aagattgact
    13801 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
    13861 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
    13921 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
    13981 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
    14041 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
    14101 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
    14161 tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc
    14221 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
    14281 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
    14341 gcctccccgc atcgacaccg agcgctgctc gaggcctcga ctgtgccttc tagttgccag
    14401 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
    14461 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
    14521 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
    14581 gctggggaca cgtgggatcc ggtgccaggg cgtgcccttg ggctccccgg gcgcgtttgg
    14641 gcggccgcat cgaattctac cgggtagggg aggcgctttt cccaaggcag tctggagcat
    14701 gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac
    14761 attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac
    14821 cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga
    14881 cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc
    14941 aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct
    15001 gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg
    15061 gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt
    15121 ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcaat cgccgctagc
    15181 cgccaccatg aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
    15241 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
    15301 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
    15361 caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct
    15421 tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
    15481 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
    15541 catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc
    15601 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
    15661 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gtaagtgcta gcccctaggc
    15721 ctgaatcgaa gttcctattc ttcaaaaggt ataggaactt cgagctcgtc gactgcagag
    15781 gcctgcatgc aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
    15841 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
    15901 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
    15961 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
    16021 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
    16081 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
    16141 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
    16201 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
    16261 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
    16321 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
    16381 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
    16441 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
    16501 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
    16561 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
    16621 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
    16681 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
    16741 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
    16801 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
    16861 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
    16921 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
    16981 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
    17041 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
    17101 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
    17161 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
    17221 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
    17281 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
    17341 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
    17401 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
    17461 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
    17521 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
    17581 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
    17641 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
    17701 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
    17761 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
    17821 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
    17881 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
    17941 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
    18001 aaaataggcg tatcacgagg ccctttcgtc
//
