LOCUS       sequence-470355-       10085 bp    DNA     circular SYN 29-APR-2025
DEFINITION  Dox-inducible expression of the catalytic-domain (CD) of epigenetic 
            effector Kmt5c fused with scFV to programme H4K20me3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10085)
  AUTHORS   Policarpi C, Munafo M, Tsagkris S, Carlini V, Hackett JA
  TITLE     Systematic epigenome editing captures the context-dependent 
            instructive function of chromatin modifications.
  JOURNAL   Nat Genet. 2024 Jun;56(6):1168-1180. doi: 
            10.1038/s41588-024-01706-w. Epub 2024 May 9.
  PUBMED    38724747
REFERENCE   2  (bases 1 to 10085)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 29, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41588-024-01706-w"; journalName: "Nat Genet"; date: 
            "2024-06"; volume: "56"; issue: "6"; pages: "1168-1180"
COMMENT     Sequence Label: pPB_TRE3G_scFV-sfGFP-Kmt5cFL
FEATURES             Location/Qualifiers
     source          1..10085
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(268..289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     480..499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     713..735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     727..744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   874..936
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        1385..1763
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    1393..1411
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1429..1447
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1465..1483
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1501..1519
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1537..1555
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1573..1591
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1609..1627
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1672..1696
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2520..2546
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2637..3347
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             2637..3347
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(2664..2692)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3287..3307
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             3366..3386
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3393..3413
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4983..5104
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             5666..6460
                     /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(5720..5739)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6330..6349
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6655..6776
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   7539..7573
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8064..8080)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8064..8080)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8077..8099)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8088..8104
                     /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(8112..8118,8119..8136,8137..8142))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8157..8178
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8295..8312)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8466..9054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8546..8565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9225..10016,10017..10085))
                     /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     9848..9867
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc acggccacta gtctttcgtc ttcaagaatt
     1381 cctcgagttt actccctatc agtgatagag aacgtatgaa gagtttactc cctatcagtg
     1441 atagagaacg tatgcagact ttactcccta tcagtgatag agaacgtata aggagtttac
     1501 tccctatcag tgatagagaa cgtatgacca gtttactccc tatcagtgat agagaacgta
     1561 tctacagttt actccctatc agtgatagag aacgtatatc cagtttactc cctatcagtg
     1621 atagagaacg tataagcttt aggcgtgtac ggtgggcgcc tataaaagca gagctcgttt
     1681 agtgaaccgt cagatcgcct ggagcaattc cacaacactt ttgtcttata ccaactttcc
     1741 gtaccacttc ctaccctcgt aaaggtctag agctagccat gggccccgac atcgtgatga
     1801 cccagagccc cagcagcctg agcgccagcg tgggcgaccg cgtgaccatc acctgccgca
     1861 gcagcaccgg cgccgtgacc accagcaact acgccagctg ggtgcaggag aagcccggca
     1921 agctgttcaa gggcctgatc ggcggcacca acaaccgcgc ccccggcgtg cccagccgct
     1981 tcagcggcag cctgatcggc gacaaggcca ccctgaccat cagcagcctg cagcccgagg
     2041 acttcgccac ctacttctgc gccctgtggt acagcaacca ctgggtgttc ggccagggca
     2101 ccaaggtgga gctgaagcgt ggaggcggag gtagtggagg tggaggctct ggtgggggag
     2161 gaagcagtgg aggcggaagt gaggtgaagc tgctggagag cggaggcgga ctggtgcagc
     2221 ccggcggatc tctgaagctg agctgcgccg tgagcggctt cagcctgacc gactacggcg
     2281 tgaactgggt gcgccaggcc cccggacgcg gactggagtg gatcggtgtg atctggggcg
     2341 acggcatcac cgactacaac agcgccctga aggaccgctt catcatcagc aaggacaacg
     2401 gcaagaacac cgtgtacctg cagatgagca aggtgcgcag cgacgacacc gccctgtact
     2461 actgcgtgac cggcctgttc gactactggg gacagggcac cctggtgacc gtgagcagct
     2521 acccatacga tgttccagat tacgctggtg gaggcggagg ttctggggga ggaggtagtg
     2581 gcggtggtgg ttcaggaggc ggcggaagct tggatccagg tggaggtgga agcggtagca
     2641 aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta
     2701 atgggcacaa attttctgtc cgtggagagg gtgaaggtga tgctacaaac ggaaaactca
     2761 cccttaaatt tatttgcact actggaaaac tacctgttcc gtggccaaca cttgtcacta
     2821 ctctgaccta tggtgttcaa tgcttttccc gttatccgga tcacatgaaa cggcatgact
     2881 ttttcaagag tgccatgccc gaaggttatg tacaggaacg cactatatct ttcaaagatg
     2941 acgggaccta caagacgcgt gctgaagtca agtttgaagg tgataccctt gttaatcgta
     3001 tcgagttaaa gggtattgat tttaaagaag atggaaacat tcttggacac aaactcgagt
     3061 acaactttaa ctcacacaat gtatacatca cggcagacaa acaaaagaat ggaatcaaag
     3121 ctaacttcaa aattcgccac aacgttgaag atggttccgt tcaactagca gaccattatc
     3181 aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat tacctgtcga
     3241 cacaatctgt cctttcgaaa gatcccaacg aaaagcgtga ccacatggtc cttcttgagt
     3301 ttgtaactgc tgctgggatt acacatggca tggatgagct ctacaaaggt ggaggtcgga
     3361 ccggccccaa gaaaaaacgc aaggtggaag atcctaagaa aaagcggaaa gtggacggca
     3421 ttggtagtgg gagcaacggc agcagcggat ccggaggcgg tggtaccggg cctgatcgag
     3481 tgacagcccg agaactgtgc gagaatgatg acctggccac cagccttgtt cttgacccct
     3541 acctcggctt ccgcacccat aagatgaatg tcagtcctgt gcccaccttg cgacgacaac
     3601 accaccttcg ttcagcactg gaagctttcc taaggcaacg ggacctggag gctgcattcc
     3661 gggccctgac gctaggaggc tggatggccc actacttcca gagccgtgcc cctcggcagg
     3721 aggctgctct caagacccac attttctgct acctccgcgc cttcctgcct gagagtggct
     3781 tcaccattct gccctgcacc cgctactcca tggagaccaa tggggccaag attgtgtcta
     3841 cccgtgcttg gaagaagaat gagaagctgg agctgctggt gggctgcatt gcagagctgc
     3901 gtgaagagga tgaagacctg cttagggccg gtgagaacga cttcagcatc atgtactcca
     3961 cccgaaaaag gagtgcccag ctgtggcttg gcccagctgc cttcatcaac catgactgca
     4021 aacccaactg caagtttgtg ccctcagatg ggaatactgc ctgtgtgaag gtgctacggg
     4081 acattgagcc aggggatgaa gtgacttgct tctatggtga gggcttcttc ggcgagaaga
     4141 atgagcactg tgaatgctat acctgtgaga ggaaaggcga gggagctttc agacttcagc
     4201 ccagggagcc tgagctgcga ccaaagcccc tagacaagta tgagctccga gagacaaaga
     4261 ggcgtctgca gcagggcttg gtcagcagcc agcagagcct gatgagccgg tgggcctgct
     4321 cccacctgtc cccactgcgc cctgacccat tctgtgctgc ctgccagccc tcatgcctgc
     4381 tgcctgccag cccccacatg gattacttgc ccctttggct ccagcgggcg cctcaacccc
     4441 agcccatagt acctcccagg aagcgccacc gccgccgccg ccccaggatc cgccaggctt
     4501 cattgcctcc tgtcctccgc actgcctgtg tccccctgca ccgatgggga ggctgtggtc
     4561 cccactgcca gctacgtgct gaggccatgg tgaccctaca cctgcgcccc cagactcgct
     4621 ggaccccaca acaggactgg tactgggccc ggcgctatgg gctaccttct gtggggcgtg
     4681 tggaacttac ccgcctagcc ccagccctgc cagctgctcc tgcccctgct gggaacccag
     4741 gtcctgtccc cactcctgat ttcatcccta agcaggccct tgcctttgct ccgttctgcc
     4801 cacctaagcg cctccggcta gtggtcagtc atggttctat tgatctggac atcaatagtg
     4861 gtgagccatg acaaaagatc ataatcagcc atatcacatc tgtagaggtt ttacttgctt
     4921 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     4981 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5041 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5101 cttatcatgt ctcatatggc cttctgcagg agcgtacaga acccagggcc ctggcacccg
     5161 tgcagaccct ggcccacccc acctgggcgc tcagtgccca agagatgtcc acacctagga
     5221 tgtcccgcgg tgggtggggg gcccgagaga cgggcaggcc gggggcaggc ctggccatgc
     5281 ggggccgaac cgggcactgc ccagcgtggg gcgcgggggc cacggcgcgc gcccccagcc
     5341 cccgggccca gcaccccaag gcggccaacg ccaaaactct ccctcctcct cttcctcaat
     5401 ctcgctctcg ctcttttttt ttttcgcaaa aggaggggag agggggtaaa aaaatgctgc
     5461 actgtgcggc gaagccggtg agtgagcggc gcggggccaa tcagcgtgcg ccgttccgaa
     5521 agttgccttt tatggctcga gcggccgcgg cggcgcccta taaaacccag cggcgcgacg
     5581 cgccaccacc gccgagaccg cgtccgcccc gcgagcacag agcctcgcct ttgccgatcc
     5641 tctagagtcg agatccgccg ccaccatgat tgaacaagat ggattgcacg caggttctcc
     5701 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     5761 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     5821 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     5881 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     5941 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     6001 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     6061 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     6121 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     6181 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     6241 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     6301 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     6361 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     6421 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     6481 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     6541 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     6601 ggggatctca tgctggagtt cttcgcccac cccccggatc taagctctag agttaacttg
     6661 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6721 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6781 gtctggaatt gactcaaatg atgtcaatta gtctatcaga agctcatctg gtctcccttc
     6841 cgggggacaa gacatccctg tttaatattt aaacagcagt gttcccaaac tgggttctta
     6901 tatcccttgc tctggtcaac caggttgcag ggtttcctgt cctcacagga acgaagtccc
     6961 taaagaaaca gtggcagcca ggtttagccc cggaattgac tggattcctt ttttagggcc
     7021 cattggtatg gctttttccc cgtatccccc caggtgtctg caggctcaaa gagcagcgag
     7081 aagcgttcag aggaaagcga tcccgtgcca ccttccccgt gcccgggctg tccccgcacg
     7141 ctgccggctc ggggatgcgg ggggagcgcc ggaccggagc ggagccccgg gcggctcgct
     7201 gctgccccct agcgggggag ggacgtaatt acatccctgg gggctttggg ggggggctgt
     7261 ccctgatatc tataacaaga aaatatatat ataataagtt atcacgtaag tagaacatga
     7321 aataacaata taattatcgt atgagttaaa tcttaaaagt cacgtaaaag ataatcatgc
     7381 gtcattttga ctcacgcggt cgttatagtt caaaatcagt gacacttacc gcattgacaa
     7441 gcacgcctca cgggagctcc aagcggcgac tgagatgtcc taaatgcaca gcgacggatt
     7501 cgcgctattt agaaagagag agcaatattt caagaatgca tgcgtcaatt ttacgcagac
     7561 tatctttcta gggttaatct agctgcatca ggatcatatc gtcgggtctt ttttccggct
     7621 cagtcatcgc ccaagctggc gctatctggg catcggggag gaagaagccc gtgccttttc
     7681 ccgcgaggtt gaagcggcat ggaaagagtt tgccgaggat gactgctgct gcattgacgt
     7741 tgagcgaaaa cgcacgttta ccatgatgat tcgggaaggt gtggccatgc acgcctttaa
     7801 cggtgaactg ttcgttcagg ccacctggga taccagttcg tcgcggcttt tccggacaca
     7861 gttccggatg gtcagcccga agcgcatcag caacccgaac aataccggcg acagccggaa
     7921 ctgccgtgcc ggtgtgcaga ttaatgacag cggtgcggcg ctgggatatt acgtcagcga
     7981 ggacgggtat cctggctgga tgccgcagaa atggacatgg ataccccgtg agttacccgg
     8041 cgggcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     8101 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     8161 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     8221 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     8281 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     8341 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     8401 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     8461 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     8521 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     8581 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     8641 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     8701 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     8761 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     8821 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     8881 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     8941 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     9001 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     9061 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     9121 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     9181 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     9241 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     9301 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     9361 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     9421 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     9481 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     9541 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     9601 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     9661 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     9721 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     9781 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     9841 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     9901 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     9961 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
    10021 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
    10081 ctcat
//
