pLenti-DsRed_IRES_MAPT:EGFP

LOCUS       pLenti-DsRed_IRES_MAPT:EGFP               11269 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Lentiviral overexpression vector to make stable bicistronic cell 
            line for Tau screen.
ACCESSION   .
VERSION     .
KEYWORDS    pLenti-DsRed_IRES_MAPT:EGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11269)
  AUTHORS   Rousseaux MW, de Haro M, Lasagna-Reeves CA, De Maio A, Park J, 
            Jafar-Nejad P, Al-Ramahi I, Sharma A, See L, Lu N, Vilanova-Velez L,
            Klisch TJ, Westbrook TF, Troncoso JC, Botas J, Zoghbi HY
  TITLE     TRIM28 regulates the nuclear accumulation and toxicity of both 
            alpha-synuclein and tau.
  JOURNAL   Elife. 2016 Oct 25;5. pii: e19809. doi: 10.7554/eLife.19809.
  PUBMED    27779468
REFERENCE   2  (bases 1 to 11269)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..11269
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             3..636
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    683..808
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1305..1538
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1723..1767
                     /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    2060..2177
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2229..2532
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2533..2736
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2687..2707
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2733..2757
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2840..2849
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2846..3523
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(2976..2994)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(3026..3046)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     3244..3263
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3438..3461
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    3549..4122
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3572..4124
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3716..3733)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3943..3962
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5477..6196
                     /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             5477..6193
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5522..5543)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5783..5802)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6130..6151
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6216..6235)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6217..6235)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6280..6303)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6665..6684
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6753..7547
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6807..6826)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7417..7436
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     misc_feature    7661..8249
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7714..7734)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8132..8143)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8324..8557
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8676..8694)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8762..8866
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8867..9727
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(9085..9104)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9898..10486
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10387..10406
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10640..10657
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     10749..10768
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     protein_bind    10897..10918
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        10933..10963
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10971..10987
                     /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)."
     primer_bind     10976..10998
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10995..11011
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10995..11011
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11061..11080
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 gttggaaggg ctaattcact cccaaagaag acaagatatc cttgatctgt ggatctacca
       61 cacacaaggc tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc
      121 actgaccttt ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc
      181 caataaagga gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc
      241 ggagagagaa gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg
      301 agagctgcat ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc
      361 cgctggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag
      421 atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct
      481 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
      541 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
      601 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
      661 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
      721 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
      781 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
      841 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
      901 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
      961 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     1021 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     1081 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     1141 gcaagcggcc ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga
     1201 agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag
     1261 gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt
     1321 gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag
     1381 gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag
     1441 gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc
     1501 ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga
     1561 aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa
     1621 cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc
     1681 ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta
     1741 ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg
     1801 tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct
     1861 gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac
     1921 ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga
     1981 gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgccg aattcacaaa
     2041 tggcagtatt catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg
     2101 aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta
     2161 caaaaattca aaattttcgg gtttattaca gggacagcag agatccagtt tggactagtg
     2221 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     2281 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     2341 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     2401 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     2461 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     2521 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     2581 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     2641 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     2701 aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcgcc
     2761 tggagacgcc atccacgctg ttttgacctc catagaagac accggcggcc tagcgctacc
     2821 ggactcagat ccaccggtcg ccaccatggc ctcctccgag gacgtcatca aggagttcat
     2881 gcgcttcaag gtgcgcatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
     2941 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg
     3001 ccccctgccc ttcgcctggg acatcctgtc cccccagttc cagtacggct ccaaggtgta
     3061 cgtgaagcac cccgccgaca tccccgacta caagaagctg tccttccccg agggcttcaa
     3121 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
     3181 cctgcaggac ggctccttca tctacaaggt gaagttcatc ggcgtgaact tcccctccga
     3241 cggccccgta atgcagaaga agactatggg ctgggaggcc tccaccgagc gcctgtaccc
     3301 ccgcgacggc gtgctgaagg gcgagatcca caaggccctg aagctgaagg acggcggcca
     3361 ctacctggtg gagttcaagt ccatctacat ggccaagaag cccgtgcagc tgcccggcta
     3421 ctactacgtg gactccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga
     3481 gcagtacgag cgcgccgagg gccgccacca cctgttcctg tagcggccaa ttctgcagtc
     3541 gacatccgcc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa
     3601 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     3661 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     3721 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     3781 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     3841 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     3901 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     3961 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     4021 cctcggtaca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     4081 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca caatcgatat
     4141 ggctgagccc cgccaggagt tcgaagtgat ggaagatcac gctgggacgt acgggttggg
     4201 ggacaggaaa gatcaggggg gctacaccat gcaccaagac caagagggtg acacggacgc
     4261 tggcctgaaa gaatctcccc tgcagacccc cactgaggac ggatctgagg aaccgggctc
     4321 tgaaacctct gatgctaaga gcactccaac agcggaagat gtgacagcac ccttagtgga
     4381 tgagggagct cccggcaagc aggctgccgc gcagccccac acggagatcc cagaaggaac
     4441 cacagctgaa gaagcaggca ttggagacac ccccagcctg gaagacgaag ctgctggtca
     4501 cgtgacccaa gctcgcatgg tcagtaaaag caaagacggg actggaagcg atgacaaaaa
     4561 agccaagggg gctgatggta aaacgaagat cgccacaccg cggggagcag cccctccagg
     4621 ccagaagggc caggccaacg ccaccaggat tccagcaaaa accccgcccg ctccaaagac
     4681 accacccagc tctggtgaac ctccaaaatc aggggatcgc agcggctaca gcagccccgg
     4741 ctccccaggc actcccggca gccgctcccg caccccgtcc cttccaaccc cacccacccg
     4801 ggagcccaag aaggtggcag tggtccgtac tccacccaag tcgccgtctt ccgccaagag
     4861 ccgcctgcag acagcccccg tgcccatgcc agacctgaag aatgtcaagt ccaagatcgg
     4921 ctccactgag aacctgaagc accagccggg aggcgggaag gtgcagataa ttaataagaa
     4981 gctggatctt agcaacgtcc agtccaagtg tggctcaaag gataatatca aacacgtccc
     5041 gggaggcggc agtgtgcaaa tagtctacaa accagttgac ctgagcaagg tgacctccaa
     5101 gtgtggctca ttaggcaaca tccatcataa accaggaggt ggccaggtgg aagtaaaatc
     5161 tgagaagctt gacttcaagg acagagtcca gtcgaagatt gggtccctgg acaatatcac
     5221 ccacgtccct ggcggaggaa ataaaaagat tgaaacccac aagctgacct tccgcgagaa
     5281 cgccaaagcc aagacagacc acggggcgga gatcgtgtac aagtcgccag tggtgtctgg
     5341 ggacacgtct ccacggcatc tcagcaatgt ctcctccacc ggcagcatcg acatggtaga
     5401 ctcgccccag ctcgccacgc tagctgacga ggtgtctgcc tccctggcca agcagggttt
     5461 gctcgaggcg gccgctatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     5521 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     5581 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     5641 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     5701 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     5761 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     5821 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     5881 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     5941 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     6001 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     6061 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     6121 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     6181 cgagctgtac aagtaatcta gataactgat cctctcccta tagtgagtcg tattaaattc
     6241 taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg
     6301 ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt
     6361 aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta
     6421 gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta
     6481 gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc
     6541 ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga
     6601 aggggtgggt ccgggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc
     6661 ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct
     6721 ccgggccttt cgacctgcag cccaagctta ccatgattga acaagatgga ttgcacgcag
     6781 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacgatcg
     6841 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     6901 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     6961 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     7021 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     7081 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     7141 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     7201 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     7261 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     7321 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     7381 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     7441 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     7501 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgacct gaggatcata
     7561 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     7621 ctgaacctga aacataaaat gaatgcaatt gatagatcct aatcaacctc tggattacaa
     7681 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     7741 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     7801 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     7861 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     7921 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     7981 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     8041 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     8101 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     8161 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     8221 tcggatctcc ctttgggccg cctccccgcc tgagatcctt taagaccaat gacttacaag
     8281 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     8341 tcccaacgaa gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     8401 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     8461 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     8521 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     8581 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggcc cgggttaatt
     8641 aaggaaaggg ctagatcatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     8701 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     8761 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     8821 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     8881 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     8941 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     9001 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     9061 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     9121 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     9181 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     9241 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     9301 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     9361 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     9421 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     9481 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     9541 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     9601 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     9661 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     9721 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     9781 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     9841 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     9901 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     9961 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
    10021 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
    10081 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
    10141 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
    10201 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
    10261 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
    10321 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
    10381 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
    10441 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
    10501 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
    10561 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
    10621 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
    10681 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agcaagctca tggctgacta
    10741 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
    10801 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tccccgtggc acgacaggtt
    10861 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
    10921 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
    10981 ataacaattt cacacaggaa acagctatga catgattacg aatttcacaa ataaagcatt
    11041 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
    11101 gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat
    11161 taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt
    11221 ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagta
//