LOCUS       sequence-221188-       17170 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Target a Cre-dependent GCaMP6s cassette and a tTA2 cassette into the
            mouse TIGRE locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17170)
  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 17170)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2018.06"; journalName: "Cell"; date: "2018-07-12- 
            12"; volume: "174"; issue: "2"; pages: "465-480.e22"
COMMENT     Sequence Label: Ai162 (TIT2L-GC6s-ICL-tTA2) targeting vector
FEATURES             Location/Qualifiers
     source          1..17170
                     /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      4717..4726
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4723..4731,4732..4749,4750..4752,4753..4785,4786..4788,
                     4789..4812,4813..4839,4840..4896,4897..4902,4903..5172,
                     5173..5190,5191..5625,5626..5631,5632..6075)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      4819..4828
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     5109..5130
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5236..5257)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5497..5516)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    6100..6688
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(6571..6582)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6719..6926
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        9456..9759
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9761..10038
                     /label=chicken beta-actin promoter
     intron          10040..11057
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     10981..11003
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     11065..11084
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    11145..11178
                     /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    11216..11264
                     /note="synthetic polyadenylation signal"
     polyA_signal    11273..11749
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(11784..11803)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    11828..11876
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    12055..12088
                     /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(12113..12730,12731..12736,12737..12772,12773..12775,
                     12776..12811,12812..12814,12815..12850,12851..12859)
                     /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    12902..13490
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(13373..13384)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    13521..13728
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    13742..13775
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        13802..14301
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(13837..13860)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     14234..14253
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     complement(14949..14965)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14949..14965)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14962..14984)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14973..14989
                     /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(14997..15003,15004..15021,15022..15027))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    15042..15063
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(15180..15197)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(15351..15939)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(15431..15450)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(16110..16901,16902..16970))
                     /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     16733..16752
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(16971..17075)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     17143..17161
                     /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 cgtgccgcca ccatgggttc tcatcatcat
     4741 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac
     4801 gatgacgata aggatctcgc caccatggtc gactcatcac gtcgtaagtg gaataagaca
     4861 ggtcacgcag tcagagctat aggtcggctg agctcactcg agaacgtcta tatcaaggcc
     4921 gacaagcaga agaacggcat caaggcgaac ttccacatcc gccacaacat cgaggacggc
     4981 ggcgtgcagc tcgcctacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     5041 ctgcccgaca accactacct gagcgtgcag tccaaacttt cgaaagaccc caacgagaag
     5101 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     5161 gagctgtaca agggcggtac cggagggagc atggtgagca agggcgagga gctgttcacc
     5221 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     5281 tccggcgagg gtgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     5341 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     5401 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     5461 gaaggctaca tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     5521 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     5581 ttcaaggagg acggcaacat cctggggcac aagctggagt acaacctgcc ggaccaactg
     5641 actgaagagc agatcgcaga atttaaagag gctttctccc tatttgacaa ggacggggat
     5701 gggacaataa caaccaagga gctggggacg gtgatgcggt ctctggggca gaaccccaca
     5761 gaagcagagc tgcaggacat gatcaatgaa gtagatgccg acggtgacgg cacaatcgac
     5821 ttccctgagt tcctgacaat gatggcaaga aaaatgaaat acagggacac ggaagaagaa
     5881 attagagaag cgttcggtgt gtttgataag gatggcaatg gctacatcag tgcagcagag
     5941 cttcgccacg tgatgacaaa ccttggagag aagttaacag atgaagaggt tgatgaaatg
     6001 atcagggaag cagacatcga tggggatggt caggtaaact acgaagagtt tgtacaaatg
     6061 atgacagcga agtaaacgcg ttacgtaaag cttgtcgata atcaacctct ggattacaaa
     6121 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     6181 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     6241 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     6301 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     6361 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     6421 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     6481 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt
     6541 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     6601 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     6661 cggatctccc tttgggccgc ctccccgcat cgacaccgag cgctgctcga ggcctcgact
     6721 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     6781 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     6841 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     6901 gaagacaata gcaggcatgc tggggacacg tgggatcctc tagtctagag ccccatcctc
     6961 actgactccg tcctggagtt ggatgagaga taatggcctt acgttgtgcc aggggagggt
     7021 cgggctggat ttagcaagat ttaccttctc caaagagcgg tgctgcagtg gcacagctgc
     7081 ccacggaggt gggggggtca ccgtccctgg aggtgatgaa gaactgtggg gatgtggcac
     7141 tgagggacat ggccagtggg cacggtgggt gggttggggt tggtcttggg gatcttggag
     7201 ggcttttcca gccttcatga tttgacgatt gtatgaacat ctacatggca attctccagc
     7261 tgcctgtccc agtcctactg acccagctgt atctctccag gcaagctctt ccaccccttc
     7321 tgcttgcatc cagacaccat caaacatgca ggctcagaca cagggaccag cagtgtctgt
     7381 ggcctttttg tgctcctctc catgctgggt tttaacttgc tctttgtcct tctatcctat
     7441 cttcttatcc ttaaggctgt tctgaacgct gtgacttgga gagtgtccca gagccctcaa
     7501 cacctgcatg tcccacgtcc atgctgtcct gcacttcctt atccccaaga tctgcctctc
     7561 cgtgatgcac tgaattggca aacatgtgtc accccagacc aacaatgtca cagcaaactc
     7621 ccccttgata ggacaagggg gaatggcttt acactgagac aggggaggtt tgggttggat
     7681 atgaggaggc agtttttccc ccagagggtg gtgacgcact gaacaggttg cccaaggagg
     7741 ctgtggatgc cccatccctg caggcattca aggccaggct ggatgtggct ctgggcagcc
     7801 tgggctgctg gttgatgacc ctgcacatag cagggggttg gatctggatg agcactgtgc
     7861 tcctttgcaa cccaggccgt tctatgattc tgtcattcta aatctctctt tcagcctaaa
     7921 gctttttccc cgtatccccc caggtgtctg caggctcaaa gagcagcgag aagcgttcag
     7981 aggaaagcga tcccgtgcca ccttccccgt gcccgggctg tccccgcacg ctgccggctc
     8041 ggggatgcgg ggggagcgcc ggaccggagc ggagccccgg gcggctcgct gctgccccct
     8101 agcgggggag ggacgtaatt acatccctgg gggctttggg ggggggctgt ccctctagag
     8161 ccccatcctc actgactccg tcctggagtt ggatgagaga taatggcctt acgttgtgcc
     8221 aggggagggt cgggctggat ttagcaagat ttaccttctc caaagagcgg tgctgcagtg
     8281 gcacagctgc ccacggaggt gggggggtca ccgtccctgg aggtgatgaa gaactgtggg
     8341 gatgtggcac tgagggacat ggccagtggg cacggtgggt gggttggggt tggtcttggg
     8401 gatcttggag ggcttttcca gccttcatga tttgacgatt gtatgaacat ctacatggca
     8461 attctccagc tgcctgtccc agtcctactg acccagctgt atctctccag gcaagctctt
     8521 ccaccccttc tgcttgcatc cagacaccat caaacatgca ggctcagaca cagggaccag
     8581 cagtgtctgt ggcctttttg tgctcctctc catgctgggt tttaacttgc tctttgtcct
     8641 tctatcctat cttcttatcc ttaaggctgt tctgaacgct gtgacttgga gagtgtccca
     8701 gagccctcaa cacctgcatg tcccacgtcc atgctgtcct gcacttcctt atccccaaga
     8761 tctgcctctc cgtgatgcac tgaattggca aacatgtgtc accccagacc aacaatgtca
     8821 cagcaaactc ccccttgata ggacaagggg gaatggcttt acactgagac aggggaggtt
     8881 tgggttggat atgaggaggc agtttttccc ccagagggtg gtgacgcact gaacaggttg
     8941 cccaaggagg ctgtggatgc cccatccctg caggcattca aggccaggct ggatgtggct
     9001 ctgggcagcc tgggctgctg gttgatgacc ctgcacatag cagggggttg gatctggatg
     9061 agcactgtgc tcctttgcaa cccaggccgt tctatgattc tgtcattcta aatctctctt
     9121 tcagcctaaa gctttttccc cgtatccccc caggtgtctg caggctcaaa gagcagcgag
     9181 aagcgttcag aggaaagcga tcccgtgcca ccttccccgt gcccgggctg tccccgcacg
     9241 ctgccggctc ggggatgcgg ggggagcgcc ggaccggagc ggagccccgg gcggctcgct
     9301 gctgccccct agcgggggag ggacgtaatt acatccctgg gggctttggg ggggggctgt
     9361 ccctctagaa gtcgacggcg cgccctgcag gaatctagtt attaatagta atcaattacg
     9421 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     9481 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     9541 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     9601 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     9661 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     9721 tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc
     9781 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt
     9841 taattatttt gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg
     9901 gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc
     9961 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag
    10021 cgaagcgcgc ggcgggcggg gagtcgctgc gacgctgcct tcgccccgtg ccccgctccg
    10081 ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg
    10141 ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt
    10201 cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga
    10261 gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc
    10321 tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc
    10381 gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca
    10441 aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg
    10501 ggctgcaacc ccccctgcac ccccctcccc gagttgctga gcacggcccg gcttcgggtg
    10561 cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg
    10621 tgggggtgcc gggcggggcg gggccgcctc gggccgggga gggctcgggg gaggggcgcg
    10681 gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat
    10741 ggtaatcgtg cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat
    10801 ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca
    10861 ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc
    10921 tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg
    10981 ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct
    11041 tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg
    11101 gcaaagaatt gatttgatac cgcgggcccc gcgtggacgg taccataact tcgtataaag
    11161 tatcctatac gaagttatcc atggctagcg gcagcctcgg agtttgaata gatagaataa
    11221 aatatcttta ttttcattcc atctgtgtgt tggttttttg tgtgagatct acgggtggca
    11281 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac
    11341 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa
    11401 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag
    11461 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc
    11521 aatctccgcc tcctgggttc aagcgattct cctgcctcag gctcccgagt tgttgggatt
    11581 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac
    11641 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc
    11701 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgcctcagct
    11761 aattgagtag gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
    11821 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
    11881 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
    11941 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
    12001 agggctcgca gccaacgtcg gggcggcagg ccgtggaagt tcctattctc tagaataact
    12061 tcgtataaag tatcctatac gaagttatac gcgcgcgtcc atttaattca ccatgtctag
    12121 actggacaag agcaaagtca taaactctgc tctggaatta ctcaatgaag tcggtatcga
    12181 aggcctgacg acaaggaaac tcgctcaaaa gctgggagtt gagcagccta ccctgtactg
    12241 gcacgtgaag aacaagcggg ccctgctcga tgccctggca atcgagatgc tggacaggca
    12301 tcatacccac ttctgccccc tggaaggcga gtcatggcaa gactttctgc ggaacaacgc
    12361 caagtcattc cgctgtgctc tcctctcaca tcgcgacggg gctaaagtgc atctcggcac
    12421 ccgcccaaca gagaaacagt acgaaaccct ggaaaatcag ctcgcgttcc tgtgtcagca
    12481 aggcttctcc ctggagaacg cactgtacgc tctgtccgcc gtgggccact ttacactggg
    12541 ctgcgtattg gaggatcagg agcatcaagt agcaaaagag gaaagagaga cacctaccac
    12601 cgattctatg cccccacttc tgagacaagc aattgagctg ttcgaccatc agggagccga
    12661 acctgccttc cttttcggcc tggaactaat catatgtggc ctggagaaac agctaaagtg
    12721 cgaaagcggc gggccggccg acgcccttga cgattttgac ttagacatgc tcccagccga
    12781 tgcccttgac gactttgacc ttgatatgct gcctgctgac gctcttgacg attttgacct
    12841 tgacatgctc cccgggtaac taagtaagga tcaaatggcg cgttacgtaa agcttgtcga
    12901 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
    12961 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
    13021 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
    13081 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
    13141 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
    13201 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
    13261 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
    13321 cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
    13381 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
    13441 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgacaccg
    13501 agcgctgctc gaggcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
    13561 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
    13621 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
    13681 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggaca cgtgggatcc
    13741 ggtgccaggg cgtgcccttg ggctccccgg gcgcgtttgg gcggccgcat cgaattctac
    13801 cgggtagggg aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg
    13861 ggcacttggc gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg
    13921 cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac cttctactcc tcccctagtc
    13981 aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca
    14041 cgtctcacta gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt
    14101 tggggcagcg gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg
    14161 ggtgggtccg ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc
    14221 tccggaggcc cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct
    14281 tcctcatctc cgggcctttc gacctgcaat cgccgctagc cgccaccatg aaaaagcctg
    14341 aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc
    14401 tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg
    14461 gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc
    14521 ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg
    14581 agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg
    14641 aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg
    14701 ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca
    14761 ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg
    14821 tgatggacga caccgtcagt gtaagtgcta gcccctaggc ctgaatcgaa gttcctattc
    14881 ttcaaaaggt ataggaactt cgagctcgtc gactgcagag gcctgcatgc aagcttggcg
    14941 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
    15001 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
    15061 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
    15121 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
    15181 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
    15241 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
    15301 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
    15361 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
    15421 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
    15481 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
    15541 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
    15601 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
    15661 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
    15721 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
    15781 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
    15841 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
    15901 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
    15961 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
    16021 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
    16081 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
    16141 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
    16201 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
    16261 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
    16321 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
    16381 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
    16441 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
    16501 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
    16561 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
    16621 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
    16681 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
    16741 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
    16801 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
    16861 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
    16921 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
    16981 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
    17041 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
    17101 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
    17161 ccctttcgtc
//
