LOCUS       sequence-450971-       13432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual fluorescent reporter for histone H2B and for exogenous Rb.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13432)
  AUTHORS   Kim S, Armand J, Safonov A, Zhang M, Soni RK, Schwartz G, McGuinness
            JE, Hibshoosh H, Razavi P, Kim M, Chandarlapaty S, Yang HW
  TITLE     Sequential activation of E2F via Rb degradation and c-Myc drives 
            resistance to CDK4/6 inhibitors in breast cancer.
  JOURNAL   Cell Rep. 2023 Nov 28;42(11):113198. doi: 
            10.1016/j.celrep.2023.113198. Epub 2023 Oct 21.
  PUBMED    37865915
REFERENCE   2  (bases 1 to 13432)
  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: "Cell Rep.";
            date: "2023-11-28"; volume: "42(11):113198. doi"; pages: " 
            10.1016/j.celrep.2023.113198. Epub 2023 Oct 21"
COMMENT     Sequence Label: pLV-H2B-iRFP670-p2a-YFP-Rb-IRES-blasticidin
FEATURES             Location/Qualifiers
     source          1..13432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    680..805
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1302..1535
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1720..1764
                     /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    2027..2142
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        2220..2431
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          2432..3370
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3327..3347
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3393..3770
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             3777..4706
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="ARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAGA
                     FFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLHR
                     HDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVML
                     YRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSRI
                     VPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYEP
                     RAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     CDS             4725..4781
                     /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             4782..5489
                     /codon_start=1
                     /product="yellow fluorescent protein optimized for FRET 
                     with CyPet (Nguyen and Daugherty, 2005)"
                     /label=YPet
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKL
                     LCTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLLEF
                     LTAAGITEGMNELY"
     misc_feature    8396..8945
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(8540..8557)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     8767..8786
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    9400..9988
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9453..9473)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9871..9882)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10195..10828
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10956..10972)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10956..10972)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10969..10991)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10980..10996
                     /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(11004..11010,11011..11028,11029..11034))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11049..11070
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11187..11204)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11358..11946)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11438..11457)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12117..12908,12909..12977))
                     /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     12740..12759
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12978..13082)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    13130..13264
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13167..13186)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13221..13240
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             join(13432,1..633)
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 ggaagggcta attcactccc aaagaagaca agatatcctt gatctgtgga tctaccacac
       61 acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact
      121 gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa
      181 taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga
      241 gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga
      301 gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc
      361 tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc
      421 ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag
      481 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
      541 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
      601 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
      661 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
      721 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
      781 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
      841 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
      901 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
      961 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     1021 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     1081 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     1141 agcggccggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
     1201 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1261 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1321 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1381 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1441 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1501 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     1561 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     1621 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     1681 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     1741 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     1801 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     1861 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     1921 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     1981 agagacagat ccattcgatt agtgaacgga tctcgacggt atcgccttta aaagaaaagg
     2041 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     2101 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     2161 cagcagagat ccagtttatc gatgaggccc tttcgtcttc actcgaggtg cccgtcagtg
     2221 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
     2281 cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
     2341 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
     2401 ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc
     2461 tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct ggctgcagta
     2521 cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc
     2581 ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg
     2641 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat
     2701 ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc
     2761 gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg
     2821 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg
     2881 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat
     2941 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg
     3001 gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg
     3061 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg
     3121 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag
     3181 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg
     3241 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct
     3301 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct
     3361 tccatttcag gtgtcgtgag ggatccgcca ccatgccaga gccagcgaag tctgctcccg
     3421 ccccgaaaaa gggctccaag aaggcggtga ctaaggcgca gaagaaaggc ggcaagaagc
     3481 gcaagcgcag ccgcaaggag agctattcca tctatgtgta caaggttctg aagcaggtcc
     3541 accctgacac cggcatttcg tccaaggcca tgggcatcat gaattcgttt gtgaacgaca
     3601 ttttcgagcg catcgcaggt gaggcttccc gcctggcgca ttacaacaag cgctcgacca
     3661 tcacctccag ggagatccag acggccgtgc gcctgctgct gcctggggag ttggccaagc
     3721 acgccgtgtc cgagggtact aaggccatca ccaagtacac cagcgctaag gatccagcgc
     3781 gtaaggtcga tctcacctcc tgcgatcgcg agccgatcca catccccggc agcattcagc
     3841 cgtgcggctg cctgctagcc tgcgacgcgc aggcggtgcg gatcacgcgc attacggaaa
     3901 atgccggcgc gttctttgga cgcgaaactc cgcgggtcgg tgagctactc gccgattact
     3961 tcggcgagac cgaagcccat gcgctgcgca acgcactggc gcagtcctcc gatccaaagc
     4021 gaccggcgct gatcttcggt tggcgcgacg gcctgaccgg ccgcaccttc gacatctcac
     4081 tgcatcgcca tgacggtaca tcgatcatcg agttcgagcc tgcggcggcc gaacaggccg
     4141 acaatccgct gcggctgacg cggcagatca tcgcgcgcac caaagaactg aagtcgctcg
     4201 aagagatggc cgcacgggtg ccgcgctatc tgcaggcgat gctcggctat caccgcgtga
     4261 tgttgtaccg cttcgcggac gacggctccg ggatggtgat cggcgaggcg aagcgcagcg
     4321 acctcgagag ctttctcggt cagcactttc cggcgtcgct ggtcccgcag caggcgcggc
     4381 tactgtactt gaagaacgcg atccgcgtgg tctcggattc gcgcggcatc agcagccgga
     4441 tcgtgcccga gcacgacgcc tccggcgccg cgctcgatct gtcgttcgcg cacctgcgca
     4501 gcatctcgcc ctgccatctc gaatttctgc ggaacatggg cgtcagcgcc tcgatgtcgc
     4561 tgtcgatcat cattgacggc acgctatggg gattgatcat ctgtcatcat tacgagccgc
     4621 gtgccgtgcc gatggcgcag cgcgtcgcgg ccgaaatgtt cgccgacttc ttatcgctgc
     4681 acttcaccgc cgcccaccac caacgcctgt acaagggttc cggagccacg aacttctctc
     4741 tgttaaagca agcaggagac gtggaagaaa atcccggtcc ttctaaaggt gaagaattat
     4801 tcactggtgt tgtcccaatt ttggttgaat tagatggtga tgttaatggt cacaaatttt
     4861 ctgtctccgg tgaaggtgaa ggtgatgcta cgtacggtaa attgacctta aaattactct
     4921 gtactactgg taaattgcca gttccatggc caaccttagt cactacttta ggttatggtg
     4981 ttcaatgttt tgctagatac ccagatcata tgaaacaaca tgactttttc aagtctgcca
     5041 tgccagaagg ttatgttcaa gaaagaacta tttttttcaa agatgacggt aactacaaga
     5101 ccagagctga agtcaagttt gaaggtgata ccttagttaa tagaatcgaa ttaaaaggta
     5161 ttgattttaa agaagatggt aacattttag gtcacaaatt ggaatacaac tataactctc
     5221 acaatgttta catcactgct gacaaacaaa agaatggtat caaagctaac ttcaaaatta
     5281 gacacaacat tgaagatggt ggtgttcaat tagctgacca ttatcaacaa aatactccaa
     5341 ttggtgatgg tccagtcttg ttaccagaca accattactt atcctatcaa tctgccttat
     5401 tcaaagatcc aaacgaaaag agagaccaca tggtcttgtt agaatttttg actgctgctg
     5461 gtattaccga gggtatgaat gaattgtacc tcgagatgcc gcccaaaacc ccccgaaaaa
     5521 cggccgccac cgccgccgct gccgccgcgg aacccccggc accgccgccg ccgccccctc
     5581 ctgaggagga cccagagcag gacagcggcc cggaggacct gcctctcgtc aggcttgagt
     5641 ttgaagaaac agaagaacct gattttactg cattatgtca gaaattaaag ataccagatc
     5701 atgtcagaga gagagcttgg ttaacttggg agaaagtttc atctgtggat gaagtattgg
     5761 gaggttatat tcaaaagaaa aaggaactgt ggggaatctg tatctttatt gcagcagttg
     5821 acctagatga gatgtcgttc acttttactg agctacagaa aaacatagaa atcagtgtcc
     5881 ataaattctt taacttacta aaagaaattg ataccagtac caaagttgat aatgctatgt
     5941 caagactgtt gaagaagtat gatgtattgt ttgcactctt cagcaaattg gaaaggacat
     6001 gtgaacttat atatttgaca caacccagca gttcgatatc tactgaaata aattctgcat
     6061 tggtgctaaa agtttcttgg atcacatttt tattagctaa aggggaagta ttacaaatgg
     6121 aagatgatct ggtgatttca tttcagttaa tgctatgtgt ccttgactat tttattaaac
     6181 tctcacctcc catgttgctc aaagaaccat ataaaacagc tgttataccc attaatggtt
     6241 cacctcgaac acccaggcga ggtcagaaca ggagtgcacg gatagcaaaa caactagaaa
     6301 atgatacaag aattattgaa gttctctgta aagaacatga atgtaatata gatgaggtga
     6361 aaaatgttta tttcaaaaat tttatacctt ttatgaattc tcttggactt gtaacatcta
     6421 atggacttcc agaggttgaa aatctttcta aacgatacga agaaatttat cttaaaaata
     6481 aagatctaga tgcaagatta tttttggatc atgataaaac tcttcagact gattctatag
     6541 acagttttga aacacagaga acaccacgaa aaagtaacct tgatgaagag gtgaatgtaa
     6601 ttcctccaca cactccagtt aggactgtta tgaacactat ccaacaatta atgatgattt
     6661 taaattcagc aagtgatcaa ccttcagaaa atctgatttc ctattttaac aactgcacag
     6721 tgaatccaaa agaaagtata ctgaaaagag tgaaggatat aggatacatc tttaaagaga
     6781 aatttgctaa agctgtggga cagggttgtg tcgaaattgg atcacagcga tacaaacttg
     6841 gagttcgctt gtattaccga gtaatggaat ccatgcttaa atcagaagaa gaacgattat
     6901 ccattcaaaa ttttagcaaa cttctgaatg acaacatttt tcatatgtct ttattggcgt
     6961 gcgctcttga ggttgtaatg gccacatata gcagaagtac atctcagaat cttgattctg
     7021 gaacagattt gtctttccca tggattctga atgtgcttaa tttaaaagcc tttgattttt
     7081 acaaagtgat cgaaagtttt atcaaagcag aaggcaactt gacaagagaa atgataaaac
     7141 atttagaacg atgtgaacat cgaatcatgg aatcccttgc atggctctca gattcacctt
     7201 tatttgatct tattaaacaa tcaaaggacc gagaaggacc aactgatcac cttgaatctg
     7261 cttgtcctct taatcttcct ctccagaata atcacactgc agcagatatg tatctttctc
     7321 ctgtaagatc tccaaagaaa aaaggttcaa ctacgcgtgt aaattctact gcaaatgcag
     7381 agacacaagc aacctcagcc ttccagaccc agaagccatt gaaatctacc tctctttcac
     7441 tgttttataa aaaagtgtat cggctagcct atctccggct aaatacactt tgtgaacgcc
     7501 ttctgtctga gcacccagaa ttagaacata tcatctggac ccttttccag cacaccctgc
     7561 agaatgagta tgaactcatg agagacaggc atttggacca aattatgatg tgttccatgt
     7621 atggcatatg caaagtgaag aatatagacc ttaaattcaa aatcattgta acagcataca
     7681 aggatcttcc tcatgctgtt caggagacat tcaaacgtgt tttgatcaaa gaagaggagt
     7741 atgattctat tatagtattc tataactcgg tcttcatgca gagactgaaa acaaatattt
     7801 tgcagtatgc ttccaccagg ccccctacct tgtcaccaat acctcacatt cctcgaagcc
     7861 cttacaagtt tcctagttca cccttacgga ttcctggagg gaacatctat atttcacccc
     7921 tgaagagtcc atataaaatt tcagaaggtc tgccaacacc aacaaaaatg actccaagat
     7981 caagaatctt agtatcaatt ggtgaatcat tcgggacttc tgagaagttc cagaaaataa
     8041 atcagatggt atgtaacagc gaccgtgtgc tcaaaagaag tgctgaagga agcaaccctc
     8101 ctaaaccact gaaaaaacta cgctttgata ttgaaggatc agatgaagca gatggaagta
     8161 aacatctccc aggagagtcc aaatttcagc agaaactggc agaaatgact tctactcgaa
     8221 cacgaatgca aaagcagaaa atgaatgata gcatggatac ctcaaacaag gaagagaaat
     8281 aagaattcct cgagggcggc cgctctagag tcgacgggcc gcggtaacaa ttgttaacta
     8341 acttaagcta gcaacggttt ccctctagcg ggatcaattc cgcccccccc ccctaacgtt
     8401 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     8461 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     8521 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     8581 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     8641 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     8701 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     8761 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     8821 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg
     8881 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     8941 taataccatg gtcatgaaaa catttaacat ttctcaacaa gatctagaat tagtagaagt
     9001 agcgacagag aagattacaa tgctttatga ggataataaa catcatgtgg gagcggcaat
     9061 tcgtacgaaa acaggagaaa tcatttcggc agtacatatt gaagcgtata taggacgagt
     9121 aactgtttgt gcagaagcca ttgcgattgg tagtgcagtt tcgaatggac aaaaggattt
     9181 tgacacgatt gtagctgtta gacaccctta ttctgacgaa gtagatagaa gtattcgagt
     9241 ggtaagtcct tgtggtatgt gtagggagtt gatttcagac tatgcaccag attgttttgt
     9301 gttaatagaa atgaatggca agttagtcaa aactacgatt gaagaactca ttccactcaa
     9361 atatacccga aattaacccg ggcggggcgc gtctggaaca atcaacctct ggattacaaa
     9421 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     9481 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     9541 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     9601 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     9661 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     9721 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     9781 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     9841 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     9901 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     9961 cggatctccc tttgggccgc ctccccgcct ggaattaatt ctgcagtcga gacctagaaa
    10021 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag
    10081 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa
    10141 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagagg ggactggaag
    10201 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag
    10261 gctacttccc tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct
    10321 ttggatggtg ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag
    10381 gagagaacac cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag
    10441 aagtgttaga gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc
    10501 atccggagta cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg
    10561 actttccagg gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca
    10621 tataagcagc tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
    10681 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
    10741 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
    10801 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
    10861 ttataacttg caaagaaatg aatatcagag agtgagaggc cttgacattg ctagcgttta
    10921 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
    10981 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
    11041 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
    11101 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
    11161 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
    11221 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
    11281 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
    11341 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
    11401 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
    11461 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
    11521 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
    11581 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
    11641 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
    11701 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
    11761 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
    11821 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
    11881 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
    11941 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
    12001 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
    12061 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
    12121 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
    12181 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
    12241 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
    12301 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
    12361 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
    12421 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
    12481 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
    12541 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
    12601 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
    12661 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
    12721 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
    12781 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
    12841 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
    12901 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
    12961 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
    13021 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
    13081 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat
    13141 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
    13201 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
    13261 gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat
    13321 taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt
    13381 ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag tt
//
