LOCUS       sequence-449911-       10110 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Lentiviral expression of AID degron tagged H3.3(K27M) and mTurquoise
            fused BlasticidinR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10110)
  AUTHORS   Sinha J, Nickels JF, Thurm AR, Ludwig CH, Archibald BN, Hinks MM, 
            Wan J, Fang D, Bintu L
  TITLE     The H3.3 K36M oncohistone disrupts the establishment of epigenetic 
            memory through loss of DNA methylation.
  JOURNAL   bioRxiv [Preprint]. 2023 Oct 14:2023.10.13.562147. doi: 
            10.1101/2023.10.13.562147.
  PUBMED    37873347
REFERENCE   2  (bases 1 to 10110)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2023 Oct 14:2023.10.13.562147. doi: 
            10.1101/2023.10.13.562147."
COMMENT     Sequence Label: pLenti-H3.3(K27M)-3AID-HA-2A-mTurquoiseBSD
FEATURES             Location/Qualifiers
     source          1..10110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             651..831
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     1340..1356
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1341..1357
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1360..1948
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1466..1477
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1875..1895
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    2004..2037
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(2057..2455)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(join(2474..3184,3185..3187,3188..3190))
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2010)"
                     /label=mTurquoise
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2516..2537)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2865..2884
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3124..3145
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(3191..3244)
                     /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             complement(3254..3280)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(3293..3319)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(3320..3523)
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Kubota et al., 2013)"
                     /label=mini-AID
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRMYK"
     CDS             complement(3530..3733)
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Kubota et al., 2013)"
                     /label=mini-AID
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRMYK"
     CDS             complement(3740..3943)
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Kubota et al., 2013)"
                     /label=mini-AID
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRMYK"
     CDS             complement(3968..3994)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(3995..4198)
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Kubota et al., 2013)"
                     /label=mini-AID
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRMYK"
     CDS             complement(4205..4408)
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Kubota et al., 2013)"
                     /label=mini-AID
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRMYK"
     CDS             complement(4415..4618)
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Kubota et al., 2013)"
                     /label=mini-AID
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRMYK"
     CDS             complement(4643..4669)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          5113..6051
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(5136..5156)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        complement(6052..6263)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    6386..6503
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(6801..6845)
                     /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    7030..7263
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    7760..7885
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             7932..8112
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(8130..8328)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(8155..8175)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        8330..8709
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     8881..8900
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        8975..9079
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9080..9148,9149..9940)
                     /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     complement(9298..9317)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgc tgctagagat
      661 tttccacact gactaaaagg gtctgaggga tctctagtta ccagagtcac acaacagacg
      721 ggcacacact acttgaagca ctcaaggcaa gctttattga ggcttaagca gtgggttccc
      781 tagttagcca gagagctccc aggctcagat ctggtctaac cagagagacc cagtacagtc
      841 cggatgcagc tctcgggcca tgtgatgaaa tgctaggcgg ctgtcaaacc tccactctaa
      901 tacttctctc tccgggtcat ccatcccatg caggctcaca gggtgtaaca agcgggtgtt
      961 ctctccttca ttggcttctt ctaccttctc ttgctcaact ggtactagct tgtagcacca
     1021 tccaaaggtc agtggatatc tgatccctgg ccctggtgtg tagttctgcc aatcagggaa
     1081 gtagccttgt gtgtggtaga tccacagatc aaggatatct tgtcttcgtt gggagtgaat
     1141 tagcccttcc agtcccccct tttcttttaa aaagtggcta agatctacag ctgccttgta
     1201 agtcattggt cttaaaggta cctgaggtgt gactggaaaa cccacctcct cctcctcttg
     1261 tgcttctagc caggcacaat cagcattggt agctgctgta ttgctacttg tgattgctcc
     1321 atgtttttct aggtctcgat cgaggtcgac ggtatcgatg cggggaggcg gcccaaaggg
     1381 agatccgact cgtctgaggg cgaaggcgaa gacgcggaag aggccgcaga gccggcagca
     1441 ggccgcggga aggaaggtcc gctggattga gggccgaagg gacgtagcag aaggacgtcc
     1501 cgcgcagaat ccaggtggca acacaggcga gcagccaagg aaaggacgat gatttccccg
     1561 acaacaccac ggaattgtca gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc
     1621 aggcggcgat gagttccgcc gtggcaatag ggagggggaa agcgaaagtc ccggaaagga
     1681 gctgacaggt ggtggcaatg ccccaaccag tgggggttgc gtcagcaaac acagtgcaca
     1741 ccacgccacg ttgcctgaca acgggccaca actcctcata aagagacagc aaccaggatt
     1801 tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac aaaggcatta
     1861 aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca gtcaatcttt
     1921 cacaaatttt gtaatccaga ggttgattat cgataagctt gatatcgaat tgtacctagt
     1981 ggaaccggaa cccttaaaca tgtataactt cgtataatgt atgctatacg aagttattag
     2041 gtccctcgac gaattcttag ccctcccaca cataaccaga gggcagcaat tcacgaatcc
     2101 caactgccgt cggctgtcca tcactgtcct tcactatggc tttgatccca ggatgcagat
     2161 cgagaagcac ctgtcggcac cgtccgcagg ggctcaagat gcccctgttc tcatttccga
     2221 tcgcgacgat acaagtcagg ttgccagctg ccgcagcagc agcagtgccc agcaccacga
     2281 gttctgcaca aggtccccca gtaaaatgat atacattgac accagtgaag atgcggccgt
     2341 cgctagagag agctgcgctg gcgacgctgt agtcttcaga gatggggatg ctgttgattg
     2401 tagccgttgc tctttcaatg agggtggatt cttcttgaga caaaggcttg gccatgctac
     2461 cgctgccgct acccttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga
     2521 actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagc ttggactggg
     2581 tgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct
     2641 ggtagtggtc ggcgagctgc acgccgccgt cctcgatgtt gtggcggatc ttgaagttgg
     2701 ccttgatgcc gttcttctgc ttgtcggcgg tgatatagac gttgtcgctg atgtagttgt
     2761 actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga
     2821 tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt
     2881 cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga
     2941 agtcgtgctg cttcatgtgg tcggggtagc gggcgaagca ctgcacgccc caggacaggg
     3001 tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg
     3061 tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt
     3121 ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct
     3181 tgctcaccat tgggccagga ttctcctcga catctccgca agttaggagg cttccccttc
     3241 cctctccaga tccagcataa tcaggcacat cataagggta tccggcgctg ccagcgtagt
     3301 caggcacatc ataggggtat ttatacattc gcagatcaat tttccgcaag tagggggctc
     3361 cgtccatact tacttttaca aatgctgccg cctcggggcc tccagaactc ttttggcaac
     3421 tgaccattac attcttgcgg tatgatctga cggggggcca accgactact tgagcttttg
     3481 ccgggggttt agctgggtct ttcggacatg cactcttctc tttggaagat ttatacatgc
     3541 ggagatctat tttcctaagg tacggggcgc cgtccatgct aactttaacg aatgctgcag
     3601 cctctggtcc gccactagac ttttgacaag ataccataac attcttgcga taggatctga
     3661 caggcggcca accaactact tgagctttgg cgggtggctt cgcagggtcc ttcgggcagg
     3721 cgctcttttc tttgctgctc ttatacatcc taaggtcgat tttcctcaag tacggcgcac
     3781 catccatgga aacctttaca aacgctgcgg cctctggacc tccgcttgat ttttgacatg
     3841 ataccataac atttttcctg tatgagcgca caggaggcca accgacgacc tgagccttcg
     3901 ccggcggctt tgcagggtct tttggacagg cacttttctc cttactcaat tggccggaaa
     3961 tcgatggtgc ataatcaggc acatcatagg ggtatttata cattcgcaga tcaattttcc
     4021 gcaagtaggg ggctccgtcc atacttactt ttacaaatgc tgccgcctcg gggcctccag
     4081 aactcttttg gcaactgacc attacattct tgcggtatga tctgacgggg ggccaaccga
     4141 ctacttgagc ttttgccggg ggtttagctg ggtctttcgg acatgcactc ttctctttgg
     4201 aagatttata catgcggaga tctattttcc taaggtacgg ggcgccgtcc atgctaactt
     4261 taacgaatgc tgcagcctct ggtccgccac tagacttttg acaagatacc ataacattct
     4321 tgcgatagga tctgacaggc ggccaaccaa ctacttgagc tttggcgggt ggcttcgcag
     4381 ggtccttcgg gcaggcgctc ttttctttgc tgctcttata catcctaagg tcgattttcc
     4441 tcaagtacgg cgcaccatcc atggaaacct ttacaaacgc tgcggcctct ggacctccgc
     4501 ttgatttttg acatgatacc ataacatttt tcctgtatga gcgcacagga ggccaaccga
     4561 cgacctgagc cttcgccggc ggctttgcag ggtcttttgg acaggcactt ttctccttac
     4621 tcaattggcc ggaaatcgat ggtgcataat caggaacatc ataaggatag ccaccgttaa
     4681 cagctctctc tccccgtatc cggcgagcca actggatgtc tttgggcatg atggtgactc
     4741 tcttagcgtg gatggcacac aggttggtat cttcgaacag acccaccagg tacgcttcgc
     4801 tagcctcctg cagcgcaccg atggctgcgc tctgaaacct caggtcggtt ttgaaatcct
     4861 gcgcgatctc cctcaccaac ctctggaagg gcagcttccg gatgagcagc tcggtcgact
     4921 tctgataacg acgaatctct cgaagcgcca cggtcccggg cctgtagcga tgaggcttct
     4981 tcaccccgcc ggtagaggga gcgctcatcc tggcggcttt cgtggccagc tgtttgcggg
     5041 gggctttccc accggtggac ttacgagcag tctgcttggt tcgggccata tttaaatgct
     5101 agctcacgac acctgaaatg gaagaaaaaa actttgaacc actgtctgag gcttgagaat
     5161 gaaccaagat ccaaactcaa aaagggcaaa ttccaaggag aattacatca agtgccaagc
     5221 tggcctaact tcagtctcca cccactcagt gtggggaaac tccatcgcat aaaacccctc
     5281 cccccaacct aaagacgacg tactccaaaa gcgcgagaac taatcgaggt gcctggacgg
     5341 cgcccggtac tccgtggagt cacatgaagc gacggctgag gacggaaagg cccttttcct
     5401 ttgtgtgggt gactcacccg cccgctctcc cgagcgccgc gtcctccatt ttgagctccc
     5461 tgcagcaggg ccgggaagcg gccatctttc cgctcacgca actggtgccg accgggccag
     5521 ccttgccgcc cagggcgggg cgatacacgg cggcgcgagg ccaggcacca gagcaggccg
     5581 gccagcttga gactaccccc gtccgattct cggtggccgc gctcgcaggc cccgcctcgc
     5641 cgaacatgtg cgctgggacg cacgggcccc gtcgccgccc gcggccccaa aaaccgaaat
     5701 accagtgtgc agatcttggc ccgcatttac aagactatct tgccagaaaa aaagcgtcgc
     5761 agcaggtcat caaaaatttt aaatggctag agacttatcg aaagcagcga gacaggcgcg
     5821 aaggtgccac cagattcgca cgcggcggcc ccagcgccca ggccaggcct caactcaagc
     5881 acgaggcgaa ggggctcctt aagcgcaagg cctcgaactc tcccacccac ttccaacccg
     5941 aagctcggga tcaagaatca cgtactgcag ccaggtggaa gtaattcaag gcacgcaagg
     6001 gccataaccc gtaaagaggc caggcccgcg ggaaccacac acggcactta cctgtgttct
     6061 ggcggcaaac ccgttgcgaa aaagaacgtt cacggcgact actgcactta tatacggttc
     6121 tcccccaccc tcgggaaaaa ggcggagcca gtacacgaca tcactttccc agtttacccc
     6181 gcgccacctt ctctaggcac cggttcaatt gccgacccct ccccccaact tctcggggac
     6241 tgtgggcgat gtgcgctctg cccactgacg ggcacatgca tggcggtaat acggttatcc
     6301 acgcggccgc ctgagtcaaa tgggatctct agagcgggcc cggtactaac caaactggat
     6361 ctctgctgtc cctgtaataa acccgaaaat tttgaatttt tgtaatttgt ttttgtaatt
     6421 ctttagtttg tatgtctgtt gctattatgt ctactattct ttcccctgca ctgtaccccc
     6481 caatcccccc ttttctttta aaattgtgga tgaatactgc catttgtctg cagaattggc
     6541 gcacgcagtg ccgatccgtt cactaatcga atggatctgt ctctgtctct ctctccacct
     6601 tcttcttcta ttccttcggg cctgtcgggt cccctcgggg ttgggaggtg ggtctgaaac
     6661 gataatggtg aatatccctg cctaactcta ttcactatag aaagtacagc aaaaactatt
     6721 cttaaaccta ccaagcctcc tactatcatt atgaataatt ttatatacca cagccaattt
     6781 gttatgttaa accaattcca caaacttgcc catttatcta attccaataa ttcttgttca
     6841 ttcttttctt gctggttttg cgattcttca attaaggagt gtattaagct tgtgtaattg
     6901 ttaatttctc tgtcccactc catccaggtc gtgtgattcc aaatctgttc cagagattta
     6961 ttactccaac tagcattcca aggcacagca gtggtgcaaa tgagttttcc agagcaaccc
     7021 caaatcccca ggagctgttg atcctttagg tatctttcca cagccaggat tcttgcctgg
     7081 agctgcttga tgccccagac tgtgagttgc aacagatgct gttgcgcctc aatagccctc
     7141 agcaaattgt tctgctgctg cactatacca gacaataatt gtctggcctg taccgtcagc
     7201 gtcattgacg ctgcgcccat agtgcttcct gctgctccca agaacccaag gaacaaagct
     7261 cctattccca ctgctctttt ttctctctgc accactcttc tctttgcctt ggtgggtgct
     7321 actcctaatg gttcaatttt tactacttta tatttatata attcacttct ccaattgtcc
     7381 ctcatatctc ctcctccagg tctgaagatc agcggccggc cgcttgctgt gcggtggtct
     7441 tacttttgtt ttgctcttcc tctatcttgt ctaaagcttc cttggtgtct tttatctcta
     7501 tcctttgatg cacacaatag agggttgcta ctgtattata taatgatcta agttcttctg
     7561 atcctgtctg aagggatggt tgtagctgtc ccagtatttg tctacagcct tctgatgttt
     7621 ctaacaggcc aggattaact gcgaatcgtt ctagctccct gcttgcccat actatatgtt
     7681 ttaatttata ttttttcttt ccccctggcc ttaaccgaat tttttcccat cgcgatctaa
     7741 ttctcccccg cttaatactg acgctctcgc acccatctct ctccttctag cctccgctag
     7801 tcaaaatttt tggcgtactc accagtcgcc gcccctcgcc tcttgccgtg cgcgcttcag
     7861 caagccgagt cctgcgtcga gagagctcct ctggtttccc tttcgctttc aagtccctgt
     7921 tcgggcgcca ctgctagaga ttttccacac tgactaaaag ggtctgaggg atctctagtt
     7981 accagagtca cacaacagac gggcacacac tacttgaagc actcaaggca agctttattg
     8041 aggcttaagc agtgggttcc ctagttagcc agagagctcc caggctcaga tctggtctaa
     8101 ccagagagac ccagtacagg caaaacgcgc tgcttatata gacctcccac cgtacacgcc
     8161 taccgcccat ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat
     8221 tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg
     8281 agtcaaaccg ctatccacgc ccattgatgt actgccaaaa ccgcatcacc atggtaatag
     8341 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg
     8401 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat
     8461 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     8521 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc
     8581 gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact
     8641 ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata
     8701 atcaatgtca acgcgtatat ctggcccgta catcgcgaag cagcgcaaaa cgcctaaccc
     8761 taagcagatt cttcatgcaa ttgtcggtca agccttgcct tgttgtagct taaattttgc
     8821 tcgcgcacta ctcagcgacc tccaacacac aagcagggag cagatactgg cttaactatg
     8881 cggcatcaga gcagattgta ctgagagtgc accatagggg atcgggagat ctcccgatcc
     8941 gtcgacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     9001 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     9061 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     9121 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     9181 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     9241 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     9301 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     9361 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     9421 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     9481 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     9541 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     9601 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     9661 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     9721 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     9781 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     9841 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     9901 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     9961 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
    10021 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
    10081 agaccccgta gaaaagatca aaggatcttc
//
