LOCUS       sequence-246501-       10602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for cyclin E/A-CDK and CDK4/6 activities.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10602)
  AUTHORS   Yang HW, Cappell SD, Jaimovich A, Liu C, Chung M, Daigh LH, Pack LR,
            Fan Y, Regot S, Covert M, Meyer T
  TITLE     Stress-mediated exit to quiescence restricted by increasing 
            persistence in CDK4/6 activation.
  JOURNAL   Elife. 2020 Apr 7;9. pii: 44571. doi: 10.7554/eLife.44571.
  PUBMED    32255427
REFERENCE   2  (bases 1 to 10602)
  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: "2020-04-7"; volume: "9. pii: 44571. doi"; pages: " 
            10.7554/eLife.44571"
COMMENT     Sequence Label: pLenti-DHB-mVenus-p2a-mCherry-CDK4KTR
FEATURES             Location/Qualifiers
     source          1..10602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    181..296
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        374..585
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          586..1524
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1481..1501
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             join(1849..1851,1852..2562)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1891..1912)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2152..2171)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2499..2520
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2572..2628
                     /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             2629..3333
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2771..2789)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3039..3058
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3720..4269
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3864..3881)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4091..4110
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    4724..5312
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4777..4797)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5195..5206)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5519..6152
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6280..6296)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6280..6296)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6293..6315)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6304..6320
                     /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(6328..6334,6335..6352,6353..6358))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6373..6394
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6511..6528)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6682..7270)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6762..6781)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7441..8232,8233..8301))
                     /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     8064..8083
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8302..8406)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    8454..8588
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8491..8510)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8545..8564
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             8756..9389
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    9436..9561
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10058..10291
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10476..10520
                     /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"
ORIGIN
        1 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
       61 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
      121 gtggagagag agacagagac agatccattc gattagtgaa cggatctcga cggtatcgcc
      181 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
      241 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg
      301 gtttattaca gggacagcag agatccagtt tatcgatgag gccctttcgt cttcactcga
      361 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
      421 gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
      481 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
      541 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt
      601 ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc
      661 acctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt
      721 tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg
      781 cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat
      841 aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat
      901 agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc
      961 ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg
     1021 ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc
     1081 gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg
     1141 tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg
     1201 cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca
     1261 gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc
     1321 tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt
     1381 ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc
     1441 ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt
     1501 caaagttttt ttcttccatt tcaggtgtcg tgaggatcca ccatgacaaa tgatgtcacc
     1561 tggagcgagg cctcttcgcc tgatgagagg acactcacct ttgctgaaag atggcaatta
     1621 tcttcacctg atggagtaga tacagatgat gatttaccaa aatcgcgagc atccaaaaga
     1681 acctgtggtg tgaatgatga tgaaagtcca agcaaaattt ttatggtggg agaatctcca
     1741 caagtgtctt ccagacttca gaatttgaga ctgaataatt taattcccag gcaacttttc
     1801 aagcccaccg ataatcaaga aactggatcc ggggcccagg gcagcggcgt gagcaagggc
     1861 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1921 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     1981 aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     2041 ggctacggcc tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     2101 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     2161 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2221 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2281 tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     2341 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag
     2401 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     2461 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     2521 accgccgccg ggatcactct cggcatggac gagctgtaca agggttccgg agccacgaac
     2581 ttctctctgt taaagcaagc aggagacgtg gaagaaaatc ccggtcctgt gagcaagggc
     2641 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     2701 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     2761 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     2821 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     2881 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     2941 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     3001 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     3061 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     3121 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     3181 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     3241 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     3301 cactccaccg gcggcatgga cgagctgtac aagagtgctg gtggtagtgc tggtggtaag
     3361 cgctcggccg gtctcaacga agacgagtcg ccatctaaga agctgccatc gccgaccgtg
     3421 tcgtcaaggc tggagcgcct gaccctccag agatcaagaa tcttgagcac tagtgatgga
     3481 agtaaacatc tcccaggaga gtccaaattt cagcagaaac tggcagaaat gacttctact
     3541 cgaacacgaa tgcaaaagca gaaaatgaat gatagcatgg atacctcaaa caaggaagag
     3601 aagtaagaat tcctcgaggg cggccgctct agagtcgacg ggccgcggta acaattgtta
     3661 actaacttaa gctagcaacg gtttccctct agcgggatca attccgcccc ccccccctaa
     3721 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     3781 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     3841 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
     3901 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg
     3961 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     4021 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     4081 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     4141 accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc
     4201 gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     4261 acgataatac catggtcatg aaaacattta acatttctca acaagatcta gaattagtag
     4321 aagtagcgac agagaagatt acaatgcttt atgaggataa taaacatcat gtgggagcgg
     4381 caattcgtac gaaaacagga gaaatcattt cggcagtaca tattgaagcg tatataggac
     4441 gagtaactgt ttgtgcagaa gccattgcga ttggtagtgc agtttcgaat ggacaaaagg
     4501 attttgacac gattgtagct gttagacacc cttattctga cgaagtagat agaagtattc
     4561 gagtggtaag tccttgtggt atgtgtaggg agttgatttc agactatgca ccagattgtt
     4621 ttgtgttaat agaaatgaat ggcaagttag tcaaaactac gattgaagaa ctcattccac
     4681 tcaaatatac ccgaaattaa cccgggcggg gcgcgtctgg aacaatcaac ctctggatta
     4741 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     4801 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     4861 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     4921 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     4981 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     5041 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     5101 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     5161 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     5221 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     5281 gagtcggatc tccctttggg ccgcctcccc gcctggaatt aattctgcag tcgagaccta
     5341 gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg
     5401 ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga
     5461 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa gaggggactg
     5521 gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca
     5581 caaggctact tccctgatta gcagaactac acaccagggc caggggtcag atatccactg
     5641 acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga agaggccaat
     5701 aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga tgacccggag
     5761 agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt ggcccgagag
     5821 ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg actttccgct
     5881 ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc cctcagatcc
     5941 tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc agatctgagc
     6001 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     6061 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     6121 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     6181 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggccttgac attgctagcg
     6241 tttaccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6301 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6361 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6421 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6481 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6541 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6601 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6661 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6721 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6781 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6841 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6901 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6961 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7021 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7081 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     7141 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7201 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7261 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7321 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7381 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7441 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7501 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7561 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7621 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7681 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7741 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7801 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7861 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7921 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7981 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8041 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8101 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8161 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8221 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8281 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8341 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8401 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcaacttgt
     8461 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     8521 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     8581 tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc accccatacc
     8641 ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat tcctctgaat
     8701 tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagttggaa
     8761 gggctaattc actcccaaag aagacaagat atccttgatc tgtggatcta ccacacacaa
     8821 ggctacttcc ctgattagca gaactacaca ccagggccag gggtcagata tccactgacc
     8881 tttggatggt gctacaagct agtaccagtt gagccagata aggtagaaga ggccaataaa
     8941 ggagagaaca ccagcttgtt acaccctgtg agcctgcatg ggatggatga cccggagaga
     9001 gaagtgttag agtggaggtt tgacagccgc ctagcatttc atcacgtggc ccgagagctg
     9061 catccggagt acttcaagaa ctgctgatat cgagcttgct acaagggact ttccgctggg
     9121 gactttccag ggaggcgtgg cctgggcggg actggggagt ggcgagccct cagatcctgc
     9181 atataagcag ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg
     9241 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     9301 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     9361 cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa
     9421 gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga
     9481 ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag
     9541 agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa
     9601 attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc
     9661 agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga
     9721 caaatactgg gacagctaca accatccctt cagacaggat cagaagaact tagatcatta
     9781 tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag
     9841 gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg
     9901 gccggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat
     9961 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga
    10021 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct
    10081 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac
    10141 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac
    10201 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg
    10261 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca
    10321 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt
    10381 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac
    10441 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat
    10501 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa
    10561 aattattcat aatgatagta ggaggcttgg taggtttaag aa
//
