LOCUS       sequence-191769-       10441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses dTomato and GCaMP6s under control of the Tet-off system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10441)
  AUTHORS   Wallace J, Lord J, Dissing-Olesen L, Stevens B, Murthy VN
  TITLE     Microglial depletion disrupts normal functional development of 
            adult-born neurons in the olfactory bulb.
  JOURNAL   Elife. 2020 Mar 9;9. pii: 50531. doi: 10.7554/eLife.50531.
  PUBMED    32150529
REFERENCE   2  (bases 1 to 10441)
  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"; journalName: "Elife."; 
            date: "2020-03-9"; volume: "9. pii: 50531. doi"; pages: " 
            10.7554/eLife.50531"
COMMENT     Sequence Label: pLenti-TRE-dTomato-GCaMP6s
FEATURES             Location/Qualifiers
     source          1..10441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        99..478
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        480..678
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             696..876
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    923..1048
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1541..1774
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1959..2003
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2301..2418
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2476..2790
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    2484..2502
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2520..2538
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2555..2573
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2591..2609
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2627..2645
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2662..2680
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2698..2716
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2763..2787
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2822..2831
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2828..3529
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(2961..2979)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3229..3248
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3423..3446
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             3548..3601
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(3602..3610,3611..3628,3629..3631,3632..3664,3665..3667,
                     3668..3691,3692..3718,3719..3775,3776..3781,3782..4051,
                     4052..4069,4070..4504,4505..4510,4511..4954)
                     /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      3698..3707
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     3988..4009
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4115..4136)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4376..4395)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4994..5582
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5047..5067)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5465..5476)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5585..5601)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5586..5602)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6107..6287
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6313..6330)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6319..6543
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6589..7017
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6676..6695)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6886..6907
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7026..7046)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7031..7360
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7211..7346
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7273..7292
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7408..7455
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7474..7848
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    7978..8099
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8015..8034)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8069..8088
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8148..8164)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8148..8164)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8161..8183)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8172..8188
                     /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(8196..8202,8203..8220,8221..8226))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8241..8262
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8379..8396)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8550..9138)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8630..8649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9309..10100,10101..10169))
                     /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     9932..9951
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10170..10274)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10349..10368)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
       61 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
      121 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      181 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      241 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      301 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      361 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      421 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      481 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      541 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      601 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      661 ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct
      721 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
      781 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
      841 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
      901 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
      961 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     1021 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     1081 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     1141 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     1201 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     1261 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     1321 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     1381 gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
     1441 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
     1501 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
     1561 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
     1621 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
     1681 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
     1741 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
     1801 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
     1861 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
     1921 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
     1981 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
     2041 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
     2101 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
     2161 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
     2221 gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa
     2281 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
     2341 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     2401 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttggttaatc
     2461 gcatgctaga gagtcgagtt tactccctat cagtgataga gaacgtatgt cgagtttact
     2521 ccctatcagt gatagagaac gatgtcgagt ttactcccta tcagtgatag agaacgtatg
     2581 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
     2641 agagaacgta tgtcgagttt atccctatca gtgatagaga acgtatgtcg agtttactcc
     2701 ctatcagtga tagagaacgt atgtcgaggt aggcgtgtac ggtgggaggc ctatataagc
     2761 agagctcgtt tagtgaaccg tcagatcgcc tggagaattc gagctcggta cccggggatc
     2821 cgccaccatg gtgagcaagg gcgaggaggt catcaaagag ttcatgcgct tcaaggtgcg
     2881 catggagggc tccatgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     2941 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc
     3001 ctgggacatc ctgtcccccc agttcatgta cggctccaag gcgtacgtga agcaccccgc
     3061 cgacatcccc gattacaaga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     3121 gaacttcgag gacggcggtc tggtgaccgt gacccaggac tcctccctgc aggacggcac
     3181 gctgatctac aaggtgaaga tgcgcggcac caacttcccc cccgacggcc ccgtaatgca
     3241 gaagaagacc atgggctggg aggcctccac cgagcgcctg tacccccgcg acggcgtgct
     3301 gaagggcgag atccaccagg ccctgaagct gaaggacggc ggccactacc tggtggagtt
     3361 caagaccatc tacatggcca agaagcccgt gcaactgccc ggctactact acgtggacac
     3421 caagctggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgagcgctc
     3481 cgagggccgc caccacctgt tcctgtacgg catggacgag ctgtacaagt atttaaatgg
     3541 cagtggtgag ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc
     3601 tatgggttct catcatcatc atcatcatgg tatggctagc atgactggtg gacagcaaat
     3661 gggtcgggat ctgtacgacg atgacgataa ggatctcgcc accatggtcg actcatcacg
     3721 tcgtaagtgg aataagacag gtcacgcagt cagagctata ggtcggctga gctcactcga
     3781 gaacgtctat atcaaggccg acaagcagaa gaacggcatc aaggcgaact tccacatccg
     3841 ccacaacatc gaggacggcg gcgtgcagct cgcctaccac taccagcaga acacccccat
     3901 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcgtgcagt ccaaactttc
     3961 gaaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     4021 gatcactctc ggcatggacg agctgtacaa gggcggtacc ggagggagca tggtgagcaa
     4081 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     4141 cggccacaag ttcagcgtgt ccggcgaggg tgagggcgat gccacctacg gcaagctgac
     4201 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     4261 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     4321 cttcaagtcc gccatgcccg aaggctacat ccaggagcgc accatcttct tcaaggacga
     4381 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     4441 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     4501 caacctgccg gaccaactga ctgaagagca gatcgcagaa tttaaagagg ctttctccct
     4561 atttgacaag gacggggatg ggacaataac aaccaaggag ctggggacgg tgatgcggtc
     4621 tctggggcag aaccccacag aagcagagct gcaggacatg atcaatgaag tagatgccga
     4681 cggtgacggc acaatcgact tccctgagtt cctgacaatg atggcaagaa aaatgaaata
     4741 cagggacacg gaagaagaaa ttagagaagc gttcggtgtg tttgataagg atggcaatgg
     4801 ctacatcagt gcagcagagc ttcgccacgt gatgacaaac cttggagaga agttaacaga
     4861 tgaagaggtt gatgaaatga tcagggaagc agacatcgat ggggatggtc aggtaaacta
     4921 cgaagagttt gtacaaatga tgacagcgaa gtgagcggcc gcgactctag aattcgatat
     4981 caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     5041 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     5101 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     5161 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     5221 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     5281 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     5341 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     5401 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     5461 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     5521 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     5581 gcatcgatac cgtcgacctc gagacctaga aaaacatgga gcaatcacaa gtagcaatac
     5641 agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt
     5701 tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     5761 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     5821 tatccttgat ctgtggatct accacacaca aggctacttc cctgattggc agaactacac
     5881 accagggcca gggatcagat atccactgac ctttggatgg tgctacaagc tagtaccagt
     5941 tgagcaagag aaggtagaag aagccaatga aggagagaac acccgcttgt tacaccctgt
     6001 gagcctgcat gggatggatg acccggagag agaagtatta gagtggaggt ttgacagccg
     6061 cctagcattt catcacatgg cccgagagct gcatccggac tgtactgggt ctctctggtt
     6121 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     6181 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     6241 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcaggg cctgtttaaa
     6301 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     6361 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     6421 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     6481 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     6541 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     6601 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     6661 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     6721 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     6781 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     6841 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     6901 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     6961 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     7021 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     7081 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     7141 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     7201 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     7261 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     7321 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     7381 atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata gtatatcggc
     7441 atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag tgccgttccg
     7501 gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc
     7561 tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc
     7621 atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc
     7681 ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc
     7741 tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc
     7801 gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca cgtgctacga
     7861 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     7921 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     7981 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     8041 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     8101 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     8161 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     8221 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     8281 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     8341 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     8401 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     8461 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     8521 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     8581 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     8641 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     8701 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     8761 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     8821 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     8881 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     8941 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     9001 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     9061 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     9121 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     9181 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     9241 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     9301 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     9361 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     9421 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     9481 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     9541 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     9601 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     9661 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     9721 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     9781 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     9841 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     9901 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     9961 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
    10021 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
    10081 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
    10141 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
    10201 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
    10261 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
    10321 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
    10381 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
    10441 c
//
