LOCUS       sequence-429651-       16543 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Target a Cre-dependent GFP and a tTA2 expression cassette into the 
            mouse TIGRE locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16543)
  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 16543)
  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: Ai140 (TIT2L-GFP-ICL-tTA2) targeting vector
FEATURES             Location/Qualifiers
     source          1..16543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(39..61)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     161..180
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     374..396
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     388..405
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     389..405
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    424..457
                     /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    2985..3066
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3010..3029
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    join(3080..3098,3099..3121,3122..3140,3141..3163,3164..3182,
                     3183..3205,3206..3224,3225..3247,3248..3266,3267..3289,
                     3290..3308,3309..3331,3332..3350)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        3383..3421
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3418..3442
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    complement(3555..3588)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3665..3713
                     /note="synthetic polyadenylation signal"
     polyA_signal    3722..4198
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     polyA_signal    4210..4671
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     protein_bind    complement(4698..4731)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      4745..4754
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4751..4753,4754..4756,4757..5470)
                     /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(4751..4753,4754..4756,4757..5467)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4796..4817)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5057..5076)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5404..5425
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5495..6083
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(5966..5977)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6114..6321
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        8845..9148
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9150..9427
                     /label=chicken beta-actin promoter
     intron          9428..10444
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     10368..10390
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     10452..10471
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    10532..10565
                     /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    10603..10651
                     /note="synthetic polyadenylation signal"
     polyA_signal    10660..11136
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(11171..11190)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    11215..11263
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    11442..11475
                     /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(11496..12113,12114..12119,12120..12155,12156..12158,
                     12159..12194,12195..12197,12198..12233,12234..12242)
                     /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    12285..12873
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(12756..12767)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    12904..13111
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    13125..13158
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        13185..13684
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(13220..13243)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     13617..13636
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     complement(14332..14348)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14332..14348)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14345..14367)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14356..14372
                     /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(14380..14386,14387..14404,14405..14410))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14425..14446
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(14563..14580)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14734..15322)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14814..14833)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(15493..16284,16285..16353))
                     /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     16116..16135
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(16354..16458)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(16526..16543,1)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
       61 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
      121 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
      181 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
      241 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
      301 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
      361 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt
      421 accgaagttc ctattcttca aatagtatag gaacttcttt cggccggcca atcgattgga
      481 tccgtttaat tcgagctcgc tagagcccca tcctcactga ctccgtcctg gagttggatg
      541 agagataatg gccttacgtt gtgccagggg agggtcgggc tggatttagc aagatttacc
      601 ttctccaaag agcggtgctg cagtggcaca gctgcccacg gaggtggggg ggtcaccgtc
      661 cctggaggtg atgaagaact gtggggatgt ggcactgagg gacatggcca gtgggcacgg
      721 tgggtgggtt ggggttggtc ttggggatct tggagggctt ttccagcctt catgatttga
      781 cgattgtatg aacatctaca tggcaattct ccagctgcct gtcccagtcc tactgaccca
      841 gctgtatctc tccaggcaag ctcttccacc ccttctgctt gcatccagac accatcaaac
      901 atgcaggctc agacacaggg accagcagtg tctgtggcct ttttgtgctc ctctccatgc
      961 tgggttttaa cttgctcttt gtccttctat cctatcttct tatccttaag gctgttctga
     1021 acgctgtgac ttggagagtg tcccagagcc ctcaacacct gcatgtccca cgtccatgct
     1081 gtcctgcact tccttatccc caagatctgc ctctccgtga tgcactgaat tggcaaacat
     1141 gtgtcacccc agaccaacaa tgtcacagca aactccccct tgataggaca agggggaatg
     1201 gctttacact gagacagggg aggtttgggt tggatatgag gaggcagttt ttcccccaga
     1261 gggtggtgac gcactgaaca ggttgcccaa ggaggctgtg gatgccccat ccctgcaggc
     1321 attcaaggcc aggctggatg tggctctggg cagcctgggc tgctggttga tgaccctgca
     1381 catagcaggg ggttggatct ggatgagcac tgtgctcctt tgcaacccag gccgttctat
     1441 gattctgtca ttctaaatct ctctttcagc ctaaagcttt ttccccgtat ccccccaggt
     1501 gtctgcaggc tcaaagagca gcgagaagcg ttcagaggaa agcgatcccg tgccaccttc
     1561 cccgtgcccg ggctgtcccc gcacgctgcc ggctcgggga tgcgggggga gcgccggacc
     1621 ggagcggagc cccgggcggc tcgctgctgc cccctagcgg gggagggacg taattacatc
     1681 cctgggggct ttgggggggg gctgtccctc tagcgagctc ggtacccaga gccccatcct
     1741 cactgactcc gtcctggagt tggatgagag ataatggcct tacgttgtgc caggggaggg
     1801 tcgggctgga tttagcaaga tttaccttct ccaaagagcg gtgctgcagt ggcacagctg
     1861 cccacggagg tgggggggtc accgtccctg gaggtgatga agaactgtgg ggatgtggca
     1921 ctgagggaca tggccagtgg gcacggtggg tgggttgggg ttggtcttgg ggatcttgga
     1981 gggcttttcc agccttcatg atttgacgat tgtatgaaca tctacatggc aattctccag
     2041 ctgcctgtcc cagtcctact gacccagctg tatctctcca ggcaagctct tccacccctt
     2101 ctgcttgcat ccagacacca tcaaacatgc aggctcagac acagggacca gcagtgtctg
     2161 tggccttttt gtgctcctct ccatgctggg ttttaacttg ctctttgtcc ttctatccta
     2221 tcttcttatc cttaaggctg ttctgaacgc tgtgacttgg agagtgtccc agagccctca
     2281 acacctgcat gtcccacgtc catgctgtcc tgcacttcct tatccccaag atctgcctct
     2341 ccgtgatgca ctgaattggc aaacatgtgt caccccagac caacaatgtc acagcaaact
     2401 cccccttgat aggacaaggg ggaatggctt tacactgaga caggggaggt ttgggttgga
     2461 tatgaggagg cagtttttcc cccagagggt ggtgacgcac tgaacaggtt gcccaaggag
     2521 gctgtggatg ccccatccct gcaggcattc aaggccaggc tggatgtggc tctgggcagc
     2581 ctgggctgct ggttgatgac cctgcacata gcagagggtt ggatctggat gagcactgtg
     2641 ctcctttgca acccaggccg ttctatgatt ctgtcattct aaatctctct ttcagcctaa
     2701 agctttttcc ccgtatcccc ccaggtgtct gcaggctcaa agagcagcga gaagcgttca
     2761 gaggaaagcg atcccgtgcc accttccccg tgcccgggct gtccccgcac gctgccggct
     2821 cggggatgcg gggggagcgc cggaccggag cggagccccg ggcggctcgc tgctgccccc
     2881 tagcggggga gggacgtaat tacatccctg ggggctttgg gggggggctg tccctctagg
     2941 ggtacccggg gatccaccgg tatcgattcg gccggccgac gtcggcagtg aaaaaaatgc
     3001 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     3061 caagtttcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca
     3121 ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga
     3181 gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt
     3241 ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt
     3301 gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag
     3361 tcgagctcgg tacccgggtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc
     3421 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
     3481 aagacaccgg gaccgatcca gcctccgcgg ccccgaattc gagctcggta cccggggatc
     3541 aaattcaatt aaatataact tcgtatagca tacattatac gaagttattc gcgatgaata
     3601 aatgaaagct tgcagatccc ctcagcacca tggctagcgg cagcctcgga gtttgaatag
     3661 atagaataaa atatctttat tttcattcca tctgtgtgtt ggttttttgt gtgagatcta
     3721 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     3781 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     3841 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     3901 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     3961 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagg ctcccgagtt
     4021 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     4081 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     4141 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     4201 gcctcagcgc tgtaagtctg cagaaattga tgatctatta aacaataaag atgtccacta
     4261 aaatggaagt ttttcctgtc atactttgtt aagaagggtg agaacagagt acctacattt
     4321 tgaatggaag gattggagct acgggggtgg gggtggggtg ggattagata aatgcctgct
     4381 ctttactgaa ggctctttac tattgcttta tgataatgtt tcatagttgg atatcataat
     4441 ttaaacaagc aaaaccaaat taagggccag ctcattcctc ccactcatga tctatagatc
     4501 tatagatctc tcgtgggatc attgtttttc tcttgattcc cactttgtgg ttctaagtac
     4561 tgtggtttcc aaatgtgtca gtttcatagc ctgaagaacg agatcagcag cctctgttcc
     4621 acatacactt cattctcagt attgttttgc caagttctaa ttccatcaga aagcttttaa
     4681 ttaactgaat agctaacata acttcgtata gcatacatta tacgaagtta tcacgcgtgc
     4741 tagcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     4801 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     4861 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     4921 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     4981 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     5041 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     5101 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     5161 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     5221 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     5281 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     5341 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     5401 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     5461 gtacaagtaa acgcgttacg taaagcttgt cgataatcaa cctctggatt acaaaatttg
     5521 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     5581 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     5641 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     5701 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     5761 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     5821 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     5881 gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg
     5941 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     6001 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     6061 ctccctttgg gccgcctccc cgcatcgaca ccgagcgctg ctcgaggcct cgactgtgcc
     6121 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     6181 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     6241 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     6301 caatagcagg catgctgggg acacgtggga tcctctagag ccccatcctc actgactccg
     6361 tcctggagtt ggatgagaga taatggcctt acgttgtgcc aggggagggt cgggctggat
     6421 ttagcaagat ttaccttctc caaagagcgg tgctgcagtg gcacagctgc ccacggaggt
     6481 gggggggtca ccgtccctgg aggtgatgaa gaactgtggg gatgtggcac tgagggacat
     6541 ggccagtggg cacggtgggt gggttggggt tggtcttggg gatcttggag ggcttttcca
     6601 gccttcatga tttgacgatt gtatgaacat ctacatggca attctccagc tgcctgtccc
     6661 agtcctactg acccagctgt atctctccag gcaagctctt ccaccccttc tgcttgcatc
     6721 cagacaccat caaacatgca ggctcagaca cagggaccag cagtgtctgt ggcctttttg
     6781 tgctcctctc catgctgggt tttaacttgc tctttgtcct tctatcctat cttcttatcc
     6841 ttaaggctgt tctgaacgct gtgacttgga gagtgtccca gagccctcaa cacctgcatg
     6901 tcccacgtcc atgctgtcct gcacttcctt atccccaaga tctgcctctc cgtgatgcac
     6961 tgaattggca aacatgtgtc accccagacc aacaatgtca cagcaaactc ccccttgata
     7021 ggacaagggg gaatggcttt acactgagac aggggaggtt tgggttggat atgaggaggc
     7081 agtttttccc ccagagggtg gtgacgcact gaacaggttg cccaaggagg ctgtggatgc
     7141 cccatccctg caggcattca aggccaggct ggatgtggct ctgggcagcc tgggctgctg
     7201 gttgatgacc ctgcacatag cagggggttg gatctggatg agcactgtgc tcctttgcaa
     7261 cccaggccgt tctatgattc tgtcattcta aatctctctt tcagcctaaa gctttttccc
     7321 cgtatccccc caggtgtctg caggctcaaa gagcagcgag aagcgttcag aggaaagcga
     7381 tcccgtgcca ccttccccgt gcccgggctg tccccgcacg ctgccggctc ggggatgcgg
     7441 ggggagcgcc ggaccggagc ggagccccgg gcggctcgct gctgccccct agcgggggag
     7501 ggacgtaatt acatccctgg gggctttggg ggggggctgt ccctctagag ccccatcctc
     7561 actgactccg tcctggagtt ggatgagaga taatggcctt acgttgtgcc aggggagggt
     7621 cgggctggat ttagcaagat ttaccttctc caaagagcgg tgctgcagtg gcacagctgc
     7681 ccacggaggt gggggggtca ccgtccctgg aggtgatgaa gaactgtggg gatgtggcac
     7741 tgagggacat ggccagtggg cacggtgggt gggttggggt tggtcttggg gatcttggag
     7801 ggcttttcca gccttcatga tttgacgatt gtatgaacat ctacatggca attctccagc
     7861 tgcctgtccc agtcctactg acccagctgt atctctccag gcaagctctt ccaccccttc
     7921 tgcttgcatc cagacaccat caaacatgca ggctcagaca cagggaccag cagtgtctgt
     7981 ggcctttttg tgctcctctc catgctgggt tttaacttgc tctttgtcct tctatcctat
     8041 cttcttatcc ttaaggctgt tctgaacgct gtgacttgga gagtgtccca gagccctcaa
     8101 cacctgcatg tcccacgtcc atgctgtcct gcacttcctt atccccaaga tctgcctctc
     8161 cgtgatgcac tgaattggca aacatgtgtc accccagacc aacaatgtca cagcaaactc
     8221 ccccttgata ggacaagggg gaatggcttt acactgagac aggggaggtt tgggttggat
     8281 atgaggaggc agtttttccc ccagagggtg gtgacgcact gaacaggttg cccaaggagg
     8341 ctgtggatgc cccatccctg caggcattca aggccaggct ggatgtggct ctgggcagcc
     8401 tgggctgctg gttgatgacc ctgcacatag cagggggttg gatctggatg agcactgtgc
     8461 tcctttgcaa cccaggccgt tctatgattc tgtcattcta aatctctctt tcagcctaaa
     8521 gctttttccc cgtatccccc caggtgtctg caggctcaaa gagcagcgag aagcgttcag
     8581 aggaaagcga tcccgtgcca ccttccccgt gcccgggctg tccccgcacg ctgccggctc
     8641 ggggatgcgg ggggagcgcc ggaccggagc ggagccccgg gcggctcgct gctgccccct
     8701 agcgggggag ggacgtaatt acatccctgg gggctttggg ggggggctgt ccctctagag
     8761 tcgacggcgc gccctgcagg aatctagtta ttaatagtaa tcaattacgg ggtcattagt
     8821 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     8881 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     8941 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     9001 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     9061 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     9121 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
     9181 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
     9241 tgcagcgatg ggggcggggg gggggggggg gcgcgcgccr ggsggggsgg ggsggggsgr
     9301 ggggsggggs ggggsgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg
     9361 aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg
     9421 gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg
     9481 ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc
     9541 ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct
     9601 gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg
     9661 tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg
     9721 tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc
     9781 ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg
     9841 gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc
     9901 cctgcacccc cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac
     9961 ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg
    10021 cggggcgggg ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag
    10081 cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga
    10141 gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg
    10201 ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg
    10261 cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg
    10321 ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg
    10381 gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct
    10441 acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattgat
    10501 ttgataccgc gggccccgcg tggacggtac cataacttcg tataaagtat cctatacgaa
    10561 gttatccatg gctagcggca gcctcggagt ttgaatagat agaataaaat atctttattt
    10621 tcattccatc tgtgtgttgg ttttttgtgt gagatctacg ggtggcatcc ctgtgacccc
    10681 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta
    10741 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg
    10801 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc
    10861 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc
    10921 tgggttcaag cgattctcct gcctcaggct cccgagttgt tgggattcca ggcatgcatg
    10981 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg
    11041 ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga
    11101 ttacaggcgt gaaccactgc tcccttccct gtccttctgc ctcagctaat tgagtagggg
    11161 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
    11221 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg
    11281 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
    11341 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
    11401 aacgtcgggg cggcaggccg tggaagttcc tattctctag aataacttcg tataaagtat
    11461 cctatacgaa gttatacgcg tccatttaat tcaccatgtc tagactggac aagagcaaag
    11521 tcataaactc tgctctggaa ttactcaatg aagtcggtat cgaaggcctg acgacaagga
    11581 aactcgctca aaagctggga gttgagcagc ctaccctgta ctggcacgtg aagaacaagc
    11641 gggccctgct cgatgccctg gcaatcgaga tgctggacag gcatcatacc cacttctgcc
    11701 ccctggaagg cgagtcatgg caagactttc tgcggaacaa cgccaagtca ttccgctgtg
    11761 ctctcctctc acatcgcgac ggggctaaag tgcatctcgg cacccgccca acagagaaac
    11821 agtacgaaac cctggaaaat cagctcgcgt tcctgtgtca gcaaggcttc tccctggaga
    11881 acgcactgta cgctctgtcc gccgtgggcc actttacact gggctgcgta ttggaggatc
    11941 aggagcatca agtagcaaaa gaggaaagag agacacctac caccgattct atgcccccac
    12001 ttctgagaca agcaattgag ctgttcgacc atcagggagc cgaacctgcc ttccttttcg
    12061 gcctggaact aatcatatgt ggcctggaga aacagctaaa gtgcgaaagc ggcgggccgg
    12121 ccgacgccct tgacgatttt gacttagaca tgctcccagc cgatgccctt gacgactttg
    12181 accttgatat gctgcctgct gacgctcttg acgattttga ccttgacatg ctccccgggt
    12241 aactaagtaa ggatcaaatg gcgcgttacg taaagcttgt cgataatcaa cctctggatt
    12301 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
    12361 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
    12421 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
    12481 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
    12541 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
    12601 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
    12661 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct
    12721 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
    12781 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
    12841 cgagtcggat ctccctttgg gccgcctccc cgcatcgaca ccgagcgctg ctcgaggcct
    12901 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
    12961 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
    13021 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
    13081 attgggaaga caatagcagg catgctgggg acacgtggga tccggtgcca gggcgtgccc
    13141 ttgggctccc cgggcgcgtt tgggcggccg catcgaattc taccgggtag gggaggcgct
    13201 tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac
    13261 aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt
    13321 ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc
    13381 cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt
    13441 gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata
    13501 gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg
    13561 gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt
    13621 ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct
    13681 ttcgacctgc aatcgccgct agccgccacc atgaaaaagc ctgaactcac cgcgacgtct
    13741 gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag
    13801 ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta
    13861 aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc
    13921 gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc
    13981 atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct
    14041 gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg
    14101 agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc
    14161 atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc
    14221 agtgtaagtg ctagccccta ggcctgaatc gaagttccta ttcttcaaaa ggtataggaa
    14281 cttcgagctc gtcgactgca gaggcctgca tgcaagcttg gcgtaatcat ggtcatagct
    14341 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
    14401 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
    14461 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
    14521 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
    14581 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
    14641 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
    14701 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
    14761 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
    14821 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
    14881 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
    14941 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
    15001 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
    15061 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
    15121 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
    15181 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
    15241 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
    15301 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
    15361 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
    15421 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
    15481 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
    15541 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
    15601 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
    15661 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
    15721 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
    15781 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
    15841 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
    15901 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
    15961 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
    16021 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
    16081 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
    16141 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
    16201 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
    16261 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
    16321 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
    16381 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
    16441 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
    16501 tattatcatg acattaacct ataaaaatag gcgtatcacg agg
//
