LOCUS       sequence-220861-       18023 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 18023)
  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 18023)
  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..18023
                     /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    2959..3040
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     2984..3003
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    join(3054..3072,3073..3095,3096..3114,3115..3137,3138..3156,
                     3157..3179,3180..3198,3199..3221,3222..3240,3241..3263,
                     3264..3282,3283..3305,3306..3324)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        3357..3395
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3392..3416
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    complement(3529..3562)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    3640..3688
                     /note="synthetic polyadenylation signal"
     polyA_signal    3697..4173
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     polyA_signal    4185..4646
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     protein_bind    complement(4673..4706)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      4720..4729
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4726..4728,4729..4731,4732..5445)
                     /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(4726..4728,4729..4731,4732..5442)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4771..4792)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5032..5051)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5379..5400
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5470..6058
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(5941..5952)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6089..6296
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        8826..9129
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9131..9408
                     /label=chicken beta-actin promoter
     intron          9410..10427
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     10351..10373
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     10435..10454
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    10515..10548
                     /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    10586..10634
                     /note="synthetic polyadenylation signal"
     polyA_signal    10643..11119
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(11154..11173)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    11198..11246
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    11420..11453
                     /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      11466..11475
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(11605..11623)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     11873..11892
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     12067..12090
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             12909..12965
                     /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(12966..13583,13584..13589,13590..13625,13626..13628,
                     13629..13664,13665..13667,13668..13703,13704..13712)
                     /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    13755..14343
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(14226..14237)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    14374..14581
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    14595..14628
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        14655..15154
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(14690..14713)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     15087..15106
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     complement(15802..15818)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(15802..15818)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(15815..15837)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    15826..15842
                     /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(15850..15856,15857..15874,15875..15880))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    15895..15916
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(16033..16050)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(16204..16792)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(16284..16303)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(16963..17754,17755..17823))
                     /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     17586..17605
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(17824..17928)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     17996..18014
                     /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 gctgtcccga gctcggtacc gccccatcct cactgactcc gtcctggagt
     1741 tggatgagag ataatggcct tacgttgtgc caggggaggg tcgggctgga tttagcaaga
     1801 tttaccttct ccaaagagcg gtgctgcagt ggcacagctg cccacggagg tgggggggtc
     1861 accgtccctg gaggtgatga agaactgtgg ggatgtggca ctgagggaca tggccagtgg
     1921 gcacggtggg tgggttgggg ttggtcttgg ggatcttgga gggcttttcc agccttcatg
     1981 atttgacgat tgtatgaaca tctacatggc aattctccag ctgcctgtcc cagtcctact
     2041 gacccagctg tatctctcca ggcaagctct tccacccctt ctgcttgcat ccagacacca
     2101 tcaaacatgc aggctcagac acagggacca gcagtgtctg tggccttttt gtgctcctct
     2161 ccatgctggg ttttaacttg ctctttgtcc ttctatccta tcttcttatc cttaaggctg
     2221 ttctgaacgc tgtgacttgg agagtgtccc agagccctca acacctgcat gtcccacgtc
     2281 catgctgtcc tgcacttcct tatccccaag atctgcctct ccgtgatgca ctgaattggc
     2341 aaacatgtgt caccccagac caacaatgtc acagcaaact cccccttgat aggacaaggg
     2401 ggaatggctt tacactgaga caggggaggt ttgggttgga tatgaggagg cagtttttcc
     2461 cccagagggt ggtgacgcac tgaacaggtt gcccaaggag gctgtggatg ccccatccct
     2521 gcaggcattc aaggccaggc tggatgtggc tctgggcagc ctgggctgct ggttgatgac
     2581 cctgcacata gcagggggtt ggatctggat gagcactgtg ctcctttgca acccaggccg
     2641 ttctatgatt ctgtcattct aaatctctct ttcagcctaa agctttttcc ccgtatcccc
     2701 ccaggtgtct gcaggctcaa agagcagcga gaagcgttca gaggaaagcg atcccgtgcc
     2761 accttccccg tgcccgggct gtccccgcac gctgccggct cggggatgcg gggggagcgc
     2821 cggaccggag cggagccccg ggcggctcgc tgctgccccc tagcggggga gggacgtaat
     2881 tacatccctg ggggctttgg gggggggctg tcccggtacc cggggatcca ccggtatcga
     2941 ttcggccggc cgacgtcggc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     3001 tgctttattt gtaaccatta taagctgcaa taaacaagtt tcgagtttac cactccctat
     3061 cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg
     3121 aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact
     3181 ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga
     3241 aaagtgaaag tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagttt
     3301 accactccct atcagtgata gagaaaagtg aaagtcgagc tcggtacccg ggtcgaggta
     3361 ggcgtgtacg gtgggaggcc tatataagca gagctcgttt agtgaaccgt cagatcgcct
     3421 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc
     3481 gcggccccga attcgagctc ggtacccggg gatcaaattc aattaaatat aacttcgtat
     3541 agcatacatt atacgaagtt attcgcgagt gaataaatga aagcttgcag atcccctcag
     3601 caccatggct agcggcagcc tcggagtttg aatagataga ataaaatatc tttattttca
     3661 ttccatctgt gtgttggttt tttgtgtgag atctacgggt ggcatccctg tgacccctcc
     3721 ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata
     3781 aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg
     3841 ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat
     3901 tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg
     3961 gttcaagcga ttctcctgcc tcaggctccc gagttgttgg gattccaggc atgcatgacc
     4021 aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg
     4081 gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta
     4141 caggcgtgaa ccactgctcc cttccctgtc cttctgcctc agcgctgtaa gtctgcagaa
     4201 attgatgatc tattaaacaa taaagatgtc cactaaaatg gaagtttttc ctgtcatact
     4261 ttgttaagaa gggtgagaac agagtaccta cattttgaat ggaaggattg gagctacggg
     4321 ggtgggggtg gggtgggatt agataaatgc ctgctcttta ctgaaggctc tttactattg
     4381 ctttatgata atgtttcata gttggatatc ataatttaaa caagcaaaac caaattaagg
     4441 gccagctcat tcctcccact catgatctat agatctatag atctctcgtg ggatcattgt
     4501 ttttctcttg attcccactt tgtggttcta agtactgtgg tttccaaatg tgtcagtttc
     4561 atagcctgaa gaacgagatc agcagcctct gttccacata cacttcattc tcagtattgt
     4621 tttgccaagt tctaattcca tcagaaagct tttaattaac tgaatagcta acataacttc
     4681 gtatagcata cattatacga agttatcacg cgtgctagcg ccaccatggt gagcaagggc
     4741 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     4801 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     4861 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     4921 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     4981 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     5041 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     5101 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     5161 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     5221 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     5281 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     5341 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     5401 accgccgccg ggatcactct cggcatggac gagctgtaca agtaaacgcg ttacgtaaag
     5461 cttgtcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     5521 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     5581 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     5641 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     5701 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     5761 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     5821 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     5881 tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     5941 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     6001 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     6061 cgacaccgag cgctgctcga ggcctcgact gtgccttcta gttgccagcc atctgttgtt
     6121 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     6181 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     6241 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggacacg
     6301 tgggatcctc tagtctagag ccccatcctc actgactccg tcctggagtt ggatgagaga
     6361 taatggcctt acgttgtgcc aggggagggt cgggctggat ttagcaagat ttaccttctc
     6421 caaagagcgg tgctgcagtg gcacagctgc ccacggaggt gggggggtca ccgtccctgg
     6481 aggtgatgaa gaactgtggg gatgtggcac tgagggacat ggccagtggg cacggtgggt
     6541 gggttggggt tggtcttggg gatcttggag ggcttttcca gccttcatga tttgacgatt
     6601 gtatgaacat ctacatggca attctccagc tgcctgtccc agtcctactg acccagctgt
     6661 atctctccag gcaagctctt ccaccccttc tgcttgcatc cagacaccat caaacatgca
     6721 ggctcagaca cagggaccag cagtgtctgt ggcctttttg tgctcctctc catgctgggt
     6781 tttaacttgc tctttgtcct tctatcctat cttcttatcc ttaaggctgt tctgaacgct
     6841 gtgacttgga gagtgtccca gagccctcaa cacctgcatg tcccacgtcc atgctgtcct
     6901 gcacttcctt atccccaaga tctgcctctc cgtgatgcac tgaattggca aacatgtgtc
     6961 accccagacc aacaatgtca cagcaaactc ccccttgata ggacaagggg gaatggcttt
     7021 acactgagac aggggaggtt tgggttggat atgaggaggc agtttttccc ccagagggtg
     7081 gtgacgcact gaacaggttg cccaaggagg ctgtggatgc cccatccctg caggcattca
     7141 aggccaggct ggatgtggct ctgggcagcc tgggctgctg gttgatgacc ctgcacatag
     7201 cagggggttg gatctggatg agcactgtgc tcctttgcaa cccaggccgt tctatgattc
     7261 tgtcattcta aatctctctt tcagcctaaa gctttttccc cgtatccccc caggtgtctg
     7321 caggctcaaa gagcagcgag aagcgttcag aggaaagcga tcccgtgcca ccttccccgt
     7381 gcccgggctg tccccgcacg ctgccggctc ggggatgcgg ggggagcgcc ggaccggagc
     7441 ggagccccgg gcggctcgct gctgccccct agcgggggag ggacgtaatt acatccctgg
     7501 gggctttggg ggggggctgt ccctctagag ccccatcctc actgactccg tcctggagtt
     7561 ggatgagaga taatggcctt acgttgtgcc aggggagggt cgggctggat ttagcaagat
     7621 ttaccttctc caaagagcgg tgctgcagtg gcacagctgc ccacggaggt gggggggtca
     7681 ccgtccctgg aggtgatgaa gaactgtggg gatgtggcac tgagggacat ggccagtggg
     7741 cacggtgggt gggttggggt tggtcttggg gatcttggag ggcttttcca gccttcatga
     7801 tttgacgatt gtatgaacat ctacatggca attctccagc tgcctgtccc agtcctactg
     7861 acccagctgt atctctccag gcaagctctt ccaccccttc tgcttgcatc cagacaccat
     7921 caaacatgca ggctcagaca cagggaccag cagtgtctgt ggcctttttg tgctcctctc
     7981 catgctgggt tttaacttgc tctttgtcct tctatcctat cttcttatcc ttaaggctgt
     8041 tctgaacgct gtgacttgga gagtgtccca gagccctcaa cacctgcatg tcccacgtcc
     8101 atgctgtcct gcacttcctt atccccaaga tctgcctctc cgtgatgcac tgaattggca
     8161 aacatgtgtc accccagacc aacaatgtca cagcaaactc ccccttgata ggacaagggg
     8221 gaatggcttt acactgagac aggggaggtt tgggttggat atgaggaggc agtttttccc
     8281 ccagagggtg gtgacgcact gaacaggttg cccaaggagg ctgtggatgc cccatccctg
     8341 caggcattca aggccaggct ggatgtggct ctgggcagcc tgggctgctg gttgatgacc
     8401 ctgcacatag cagggggttg gatctggatg agcactgtgc tcctttgcaa cccaggccgt
     8461 tctatgattc tgtcattcta aatctctctt tcagcctaaa gctttttccc cgtatccccc
     8521 caggtgtctg caggctcaaa gagcagcgag aagcgttcag aggaaagcga tcccgtgcca
     8581 ccttccccgt gcccgggctg tccccgcacg ctgccggctc ggggatgcgg ggggagcgcc
     8641 ggaccggagc ggagccccgg gcggctcgct gctgccccct agcgggggag ggacgtaatt
     8701 acatccctgg gggctttggg ggggggctgt ccctctagaa gtcgacggcg cgccctgcag
     8761 gaatctagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     8821 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     8881 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     8941 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     9001 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     9061 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     9121 attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
     9181 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
     9241 gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg
     9301 cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
     9361 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg gagtcgctgc
     9421 gacgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct
     9481 gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta
     9541 attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg
     9601 ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt
     9661 gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg
     9721 gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc
     9781 cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg
     9841 ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc
     9901 gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc
     9961 tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc
    10021 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag
    10081 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
    10141 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg
    10201 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg
    10261 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg
    10321 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
    10381 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
    10441 tgctggttat tgtgctgtct catcattttg gcaaagaatt gatttgatac cgcgggcccc
    10501 gcgtggacgg taccataact tcgtataaag tatcctatac gaagttatcc atggctagcg
    10561 gcagcctcgg agtttgaata gatagaataa aatatcttta ttttcattcc atctgtgtgt
    10621 tggttttttg tgtgagatct acgggtggca tccctgtgac ccctccccag tgcctctcct
    10681 ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat
    10741 cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag
    10801 caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg
    10861 gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct
    10921 cctgcctcag gctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt
    10981 tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta
    11041 atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac
    11101 tgctcccttc cctgtccttc tgcctcagct aattgagtag gggggaggct aactgaaaca
    11161 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
    11221 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
    11281 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
    11341 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
    11401 cctcgcgagg gtttctagaa taacttcgta taaagtatcc tatacgaagt tatacgcgta
    11461 ctagcgccac catggtgagc aagggcgagg aggtcatcaa agagttcatg cgcttcaagg
    11521 tgcgcatgga gggctccatg aacggccacg agttcgagat cgagggcgag ggcgagggcc
    11581 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct
    11641 tcgcctggga catcctgtcc ccccagttca tgtacggctc caaggcgtac gtgaagcacc
    11701 ccgccgacat ccccgattac aagaagctgt ccttccccga gggcttcaag tgggagcgcg
    11761 tgatgaactt cgaggacggc ggtctggtga ccgtgaccca ggactcctcc ctgcaggacg
    11821 gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt cccccccgac ggccccgtaa
    11881 tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg
    11941 tgctgaaggg cgagatccac caggccctga agctgaagga cggcggccac tacctggtgg
    12001 agttcaagac catctacatg gccaagaagc ccgtgcaact gcccggctac tactacgtgg
    12061 acaccaagct ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgagc
    12121 gctccgaggg ccgccaccac ctgttcctgg ggcatggtac cggcagcacc ggcagcggca
    12181 gctccggcac cgcctcctcc gaggacaaca acatggccgt gattaaggag tttatgaggt
    12241 ttaaagtcag gatggagggc agcatgaacg ggcacgagtt tgagattgaa ggagaggggg
    12301 aagggaggcc ctatgagggc acccagacag caaagctgaa agtgacaaag ggcggccctc
    12361 tgccattcgc atgggacatc ctgtctcctc agtttatgta tggaagtaaa gcctacgtga
    12421 aacaccctgc tgacattcca gattacaaga aactgagctt cccagagggg tttaagtggg
    12481 aaagagtgat gaacttcgaa gacgggggcc tggtgactgt cacccaggac agctctctgc
    12541 aggacggcac tctgatctat aaggtgaaaa tgcgcggcac caattttccc cctgatggcc
    12601 cagtcatgca gaagaagaca atgggctggg aggcttcaac agaaaggctg taccccagag
    12661 acggcgtgct gaagggagag atccaccagg cactgaagct gaaagatgga gggcattatc
    12721 tggtcgaatt caaaaccatc tacatggcca agaaacccgt gcagctgcct ggctactatt
    12781 acgtcgacac aaagctggat attactagcc ataacgaaga ctacaccatt gtggagcagt
    12841 atgagcggag cgaggggagg caccatctgt ttctgtacgg catggacgag ctgtacaagg
    12901 gaagcggagc tactaacttc agcctgctga agcaggctgg agacgtggag gagaaccctg
    12961 gacctatgtc tagactggac aagagcaaag tcataaactc tgctctggaa ttactcaatg
    13021 aagtcggtat cgaaggcctg acgacaagga aactcgctca aaagctggga gttgagcagc
    13081 ctaccctgta ctggcacgtg aagaacaagc gggccctgct cgatgccctg gcaatcgaga
    13141 tgctggacag gcatcatacc cacttctgcc ccctggaagg cgagtcatgg caagactttc
    13201 tgcggaacaa cgccaagtca ttccgctgtg ctctcctctc acatcgcgac ggggctaaag
    13261 tgcatctcgg cacccgccca acagagaaac agtacgaaac cctggaaaat cagctcgcgt
    13321 tcctgtgtca gcaaggcttc tccctggaga acgcactgta cgctctgtcc gccgtgggcc
    13381 actttacact gggctgcgta ttggaggatc aggagcatca agtagcaaaa gaggaaagag
    13441 agacacctac caccgattct atgcccccac ttctgagaca agcaattgag ctgttcgacc
    13501 atcagggagc cgaacctgcc ttccttttcg gcctggaact aatcatatgt ggcctggaga
    13561 aacagctaaa gtgcgaaagc ggcgggccgg ccgacgccct tgacgatttt gacttagaca
    13621 tgctcccagc cgatgccctt gacgactttg accttgatat gctgcctgct gacgctcttg
    13681 acgattttga ccttgacatg ctccccgggt aactaagtaa ggatcaaatg gcgcgttacg
    13741 taaagcttgt cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
    13801 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
    13861 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
    13921 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
    13981 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
    14041 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
    14101 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
    14161 ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg
    14221 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
    14281 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
    14341 cgcatcgaca ccgagcgctg ctcgaggcct cgactgtgcc ttctagttgc cagccatctg
    14401 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
    14461 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
    14521 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
    14581 acacgtggga tccggtgcca gggcgtgccc ttgggctccc cgggcgcgtt tgggcggccg
    14641 catcgaattc taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt
    14701 agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac
    14761 atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac
    14821 tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca
    14881 aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa
    14941 gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca
    15001 gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg
    15061 cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc
    15121 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc aatcgccgct agccgccacc
    15181 atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac
    15241 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
    15301 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat
    15361 cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt
    15421 ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg
    15481 caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat
    15541 gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga
    15601 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat
    15661 cactggcaaa ctgtgatgga cgacaccgtc agtgtaagtg ctagccccta ggcctgaatc
    15721 gaagttccta ttcttcaaaa ggtataggaa cttcgagctc gtcgactgca gaggcctgca
    15781 tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
    15841 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
    15901 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
    15961 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
    16021 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
    16081 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
    16141 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
    16201 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
    16261 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
    16321 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
    16381 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
    16441 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
    16501 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
    16561 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
    16621 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
    16681 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
    16741 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
    16801 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
    16861 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
    16921 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
    16981 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
    17041 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
    17101 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
    17161 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
    17221 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
    17281 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
    17341 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
    17401 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
    17461 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
    17521 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
    17581 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
    17641 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
    17701 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
    17761 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
    17821 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
    17881 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
    17941 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
    18001 gcgtatcacg aggccctttc gtc
//
