LOCUS       Exported               10317 bp ds-DNA     circular SYN 27-FEB-2025
DEFINITION  Dox-inducible expression of the engineered catalytic-domain (CD) of 
            epigenetic effector Dnmt3a fused with Dnmt3l and scFV-sfGFP to 
            programme DNA methylation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10317)
  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;56:1168-1180
  PUBMED    38724747
REFERENCE   2  (bases 1 to 10317)
  AUTHORS   Epitome Tx
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Feb 28, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Under patent. Not for commercial usage or modification.
FEATURES             Location/Qualifiers
     source          1..10317
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(870..1104)
                     /label=Piggy 3'
     promoter        1386..1764
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    1394..1412
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1430..1448
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1466..1484
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1502..1520
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1538..1556
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1574..1592
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1610..1628
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1763..1798
                     /label=Fragment 1.FOR (2)
     CDS             1780..2481
                     /codon_start=1
                     /label=Recoded scFV
                     /translation="MGPDIVMTQSPSSLSASVGDRVTITCRSSTGAVTTSNYASWVQEK
                     PGKLFKGLIGGTNNRAPGVPSRFSGSLIGDKATLTISSLQPEDFATYFCALWYSNHWVF
                     GQGTKVELKRGGGGSGGGGSGGGGSSGGGSEVKLLESGGGLVQPGGSLKLSCAVSGFSL
                     TDYGVNWVRQAPGRGLEWIGVIWGDGITDYNSALKDRFIISKDNGKNTVYLQMSKVRSD
                     DTALYYCVTGLF"
     primer_bind     complement(2171..2190)
                     /label=scFV-recodedgenot_R
     CDS             2521..2547
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2548..2637
                     /codon_start=1
                     /label=linker
                     /translation="GGGGGSGGGGSGGGGSGGGGSLDPGGGGSG"
     CDS             2638..3348
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             3367..3387
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3394..3414
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    3427..3462
                     /label=Linker
     primer_bind     complement(3447..3470)
                     /label=Fragment 1.REV (2)
     primer_bind     3449..3480
                     /label=OJH599_SV40-Dnmt3a3L-scFValone.FOR
     primer_bind     3454..3491
                     /label=3a3L_KRAB_OJH091
     CDS             3469..4371
                     /codon_start=1
                     /label=Dnmt3a CD
                     /translation="NHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATGLLVLKDLGIQ
                     VDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVIGGSPCNDLSI
                     VNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVSDKRDISRFLE
                     SNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGRIAKFSKVRTI
                     TTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSRLARQRLLGRS
                     WSVPVIRHLFAPLKEYFACV"
     misc_feature    3625..3627
                     /label=E 660 A
     misc_feature    3901..3903
                     /label=E to A for catalytic mutant
     primer_bind     4325..4342
                     /label=Dnmt3a_qPCR_Fw
     CDS             4372..4452
                     /codon_start=1
                     /label=Linker
                     /translation="SSGNSNANSRGPSFSSGLVPLSLRGSH"
     CDS             4453..5097
                     /codon_start=1
                     /label=Dnmt3L
                     /translation="MGPMEIYKTVSAWKRQPVRVLSLFRNIDKVLKSLGFLESGSGSGG
                     GTLKYVEDVTNVVRRDVEKWGPFDLVYGSTQPLGSSCDRCPGWYMFQFHRILQYALPRQ
                     ESQRPFFWIFMDNLLLTEDDQETTTRFLQTEAVTLQDVRGRDYQNAMRVWSNIPGLKSK
                     HAPLTPKEEEYLQAQVRSRSKLDAPKVDLLVKNCLLPLREYFKYFSQNSLPL"
     primer_bind     complement(4490..4507)
                     /label=Dnmt3L_qPCR_rv
     primer_bind     complement(5075..5101)
                     /label=3a3L_TRE_OJH092
     primer_bind     complement(5075..5100)
                     /label=OJH_366 3a3L-pB_R
     primer_bind     complement(5078..5120)
                     /label=OJH600_Dnmt3a3L-scFValone.REV
     primer_bind     5101..5141
                     /label=Fragment 2.FOR
     primer_bind     complement(5101..5116)
                     /label=Fragment 1.REV (2)
     misc_feature    5109..6884
     primer_bind     complement(5214..5231)
                     /label=Fragment 2.REV
     polyA_signal    5217..5338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    5349..6884
     misc_feature    5349..6884
     misc_feature    5353..5873
                     /label=human beta actin promoter
     misc_feature    complement(5628..5647)
                     /label=bact-rev
     misc_feature    5628..5647
                     /label=bact-fwd
     misc_feature    5749..5768
                     /label=betact5
     CDS             5899..6693
                     /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"
     misc_feature    6024..6043
                     /label=neoseq-fwd
     misc_feature    6068..6087
                     /label=neoPstI_5
     misc_feature    complement(6134..6153)
                     /label=neoseq-rev
     misc_feature    complement(6134..6153)
                     /label=neoseq-rev
     misc_feature    6405..6423
                     /label=neoSphI_5
     misc_feature    6563..6582
                     /label=neoseq2-fwd
     misc_feature    6698..6717
                     /label=neoBstB1_5
     primer_bind     complement(6862..6902)
                     /label=Fragment 2.REV
     polyA_signal    6888..7009
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    complement(7500..7809)
                     /label='Piggy 5'
     primer_bind     complement(8296..8312)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8320..8336
                     /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(8344..8374)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8389..8410
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(8698..9286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9457..10317)
                     /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"
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 ataatcatgc gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaaa caaaaactca
     1081 aaatttcttc tataaagtaa caaaactttt atgagggaca gccccccccc aaagccccca
     1141 gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct
     1201 ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg
     1261 gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag
     1321 acacctgggg ggatacgggg aaaaggcctc cacggccact agtctttcgt cttcaagaat
     1381 tcctcgagtt tactccctat cagtgataga gaacgtatga agagtttact ccctatcagt
     1441 gatagagaac gtatgcagac tttactccct atcagtgata gagaacgtat aaggagttta
     1501 ctccctatca gtgatagaga acgtatgacc agtttactcc ctatcagtga tagagaacgt
     1561 atctacagtt tactccctat cagtgataga gaacgtatat ccagtttact ccctatcagt
     1621 gatagagaac gtataagctt taggcgtgta cggtgggcgc ctataaaagc agagctcgtt
     1681 tagtgaaccg tcagatcgcc tggagcaatt ccacaacact tttgtcttat accaactttc
     1741 cgtaccactt cctaccctcg taaaggtcta gagctagcca tgggccccga catcgtgatg
     1801 acccagagcc ccagcagcct gagcgccagc gtgggcgacc gcgtgaccat cacctgccgc
     1861 agcagcaccg gcgccgtgac caccagcaac tacgccagct gggtgcagga gaagcccggc
     1921 aagctgttca agggcctgat cggcggcacc aacaaccgcg cccccggcgt gcccagccgc
     1981 ttcagcggca gcctgatcgg cgacaaggcc accctgacca tcagcagcct gcagcccgag
     2041 gacttcgcca cctacttctg cgccctgtgg tacagcaacc actgggtgtt cggccagggc
     2101 accaaggtgg agctgaagcg tggaggcgga ggtagtggag gtggaggctc tggtggggga
     2161 ggaagcagtg gaggcggaag tgaggtgaag ctgctggaga gcggaggcgg actggtgcag
     2221 cccggcggat ctctgaagct gagctgcgcc gtgagcggct tcagcctgac cgactacggc
     2281 gtgaactggg tgcgccaggc ccccggacgc ggactggagt ggatcggtgt gatctggggc
     2341 gacggcatca ccgactacaa cagcgccctg aaggaccgct tcatcatcag caaggacaac
     2401 ggcaagaaca ccgtgtacct gcagatgagc aaggtgcgca gcgacgacac cgccctgtac
     2461 tactgcgtga ccggcctgtt cgactactgg ggccagggca ccctggtgac cgtgagcagc
     2521 tacccatacg atgttccaga ttacgctggt ggaggcggag gttctggggg aggaggtagt
     2581 ggcggtggtg gttcaggagg cggcggaagc ttggatccag gtggaggtgg aagcggtagc
     2641 aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt
     2701 aatgggcaca aattttctgt ccgtggagag ggtgaaggtg atgctacaaa cggaaaactc
     2761 acccttaaat ttatttgcac tactggaaaa ctacctgttc cgtggccaac acttgtcact
     2821 actctgacct atggtgttca atgcttttcc cgttatccgg atcacatgaa acggcatgac
     2881 tttttcaaga gtgccatgcc cgaaggttat gtacaggaac gcactatatc tttcaaagat
     2941 gacgggacct acaagacgcg tgctgaagtc aagtttgaag gtgataccct tgttaatcgt
     3001 atcgagttaa agggtattga ttttaaagaa gatggaaaca ttcttggaca caaactcgag
     3061 tacaacttta actcacacaa tgtatacatc acggcagaca aacaaaagaa tggaatcaaa
     3121 gctaacttca aaattcgcca caacgttgaa gatggttccg ttcaactagc agaccattat
     3181 caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcg
     3241 acacaatctg tcctttcgaa agatcccaac gaaaagcgtg accacatggt ccttcttgag
     3301 tttgtaactg ctgctgggat tacacatggc atggatgagc tctacaaagg tggaggtcgg
     3361 accggcccca agaaaaaacg caaggtggaa gatcctaaga aaaagcggaa agtggacggc
     3421 attggtagtg ggagcaacgg cagcagcgga tccggaggcg gtggtaccaa ccatgaccag
     3481 gaatttgacc ccccaaaggt ttacccacct gtgccagctg agaagaggaa gcccatccgc
     3541 gtgctgtctc tctttgatgg gattgctaca gggctcctgg tgctgaagga cctgggcatc
     3601 caagtggacc gctacattgc ctccgaggtg tgtgaggact ccatcacggt gggcatggtg
     3661 cggcaccagg gaaagatcat gtacgtcggg gacgtccgca gcgtcacaca gaagcatatc
     3721 caggagtggg gcccattcga cctggtgatt ggaggcagtc cctgcaatga cctctccatt
     3781 gtcaaccctg cccgcaaggg actttatgag ggtactggcc gcctcttctt tgagttctac
     3841 cgcctcctgc atgatgcgcg gcccaaggag ggagatgatc gccccttctt ctggctcttt
     3901 gagaatgtgg tggccatggg cgttagtgac aagagggaca tctcgcgatt tcttgagtct
     3961 aaccccgtga tgattgacgc caaagaagtg tctgctgcac acagggcccg ttacttctgg
     4021 ggtaaccttc ctggcatgaa caggcctttg gcatccactg tgaatgataa gctggagctg
     4081 caagagtgtc tggagcacgg cagaatagcc aagttcagca aagtgaggac cattaccacc
     4141 aggtcaaact ctataaagca gggcaaagac cagcatttcc ccgtcttcat gaacgagaag
     4201 gaggacatcc tgtggtgcac tgaaatggaa agggtgtttg gcttccccgt ccactacaca
     4261 gacgtctcca acatgagccg cttggcgagg cagagactgc tgggccgatc gtggagcgtg
     4321 ccggtcatcc gccacctctt cgctccgctg aaggaatatt ttgcttgtgt gtctagcggc
     4381 aatagtaacg ctaacagccg cgggccgagc ttcagcagcg gcctggtgcc gttaagcttg
     4441 cgcggcagcc atatgggccc tatggagata tacaagacag tgtctgcatg gaagagacag
     4501 ccagtgcggg tactgagcct ttttagaaat attgataaag tactaaagag tttgggcttt
     4561 ttagaaagcg gttctggttc tgggggagga acgctgaagt acgtggaaga tgtcacaaat
     4621 gtcgtgagga gagacgtgga gaaatggggc ccctttgacc tggtgtacgg ctcgacgcag
     4681 cccctaggca gctcctgtga tcgctgtccc ggctggtaca tgttccagtt ccaccggatc
     4741 ctgcagtatg cgctgcctcg ccaggagagt cagcggccct tcttctggat attcatggac
     4801 aatctgctgc tgactgagga tgaccaagag acaactaccc gcttccttca gacagaggct
     4861 gtgaccctcc aggatgtccg tggcagggac taccagaatg ctatgcgggt gtggagcaac
     4921 attccagggc tgaagagcaa gcatgcgccc ctgaccccaa aggaagaaga gtatctgcaa
     4981 gcccaagtca gaagcaggag caagctggac gccccgaaag ttgacctcct ggtgaagaac
     5041 tgccttctcc cgctgagaga gtacttcaag tatttttctc aaaactcact tcctctttag
     5101 ggtaccaaaa gatcataatc agccatatca catctgtaga ggttttactt gctttaaaaa
     5161 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     5221 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5281 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5341 atgtctcata tggccttctg caggagcgta cagaacccag ggccctggca cccgtgcaga
     5401 ccctggccca ccccacctgg gcgctcagtg cccaagagat gtccacacct aggatgtccc
     5461 gcggtgggtg gggggcccga gagacgggca ggccgggggc aggcctggcc atgcggggcc
     5521 gaaccgggca ctgcccagcg tgggcgcggg ggccacggcg cgcgccccca gcccccgggc
     5581 ccagcacccc aaggcggcca acgccaaaac tctccctcct cctcttcctc aatctcgctc
     5641 tcgctctttt tttttttcgc aaaaggaggg gagagggggt aaaaaaatgc tgcactgtgc
     5701 ggcgaagccg gtgagtgagc ggcgcggggc caatcagcgt gcgccgttcc gaaagttgcc
     5761 ttttatggct cgagcggccg cggcggcgcc ctataaaacc cagcggcgcg acgcgccacc
     5821 accgccgaga ccgcgtccgc cccgcgagca cagagcctcg cctttgccga tcctctagag
     5881 tcgagatccg ccgccaccat gattgaacaa gatggattgc acgcaggttc tccggccgct
     5941 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     6001 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     6061 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     6121 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     6181 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     6241 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     6301 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     6361 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6421 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     6481 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     6541 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     6601 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     6661 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     6721 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     6781 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     6841 tcatgctgga gttcttcgcc caccccccgg atctaagctc tagagttaac ttgtttattg
     6901 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6961 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     7021 attgactcaa atgatgtcaa ttagtctatc agaagctatc tggtctccct tccgggggac
     7081 aagacatccc tgtttaatat ttaaacagca gtgttcccaa actgggttct tatatccctt
     7141 gctctggtca accaggttgc agggtttcct gtcctcacag gaacgaagtc cctaaagaaa
     7201 cagtggcagc caggtttagc cccggaattg actggattcc ttttttaggg cccattggta
     7261 tggctttttc cccgtatccc cccaggtgtc tgcaggctca aagagcagcg agaagcgttc
     7321 agaggaaagc gatcccgtgc caccttcccc gtgcccgggc tgtccccgca cgctgccggc
     7381 tcggggatgc ggggggagcg ccggaccgga gcggagcccc gggcggctcg ctgctgcccc
     7441 ctagcggggg agggacgtaa ttacatccct gggggctttg ggggggggct gtccctgata
     7501 tctataacaa gaaaatatat atataataag ttatcacgta agtagaacat gaaataacaa
     7561 tataattatc gtatgagtta aatcttaaaa gtcacgtaaa agataatcat gcgtcatttt
     7621 gactcacgcg gtcgttatag ttcaaaatca gtgacactta ccgcattgac aagcacgcct
     7681 cacgggagct ccaagcggcg actgagatgt cctaaatgca cagcgacgga ttcgcgctat
     7741 ttagaaagag agagcaatat ttcaagaatg catgcgtcaa ttttacgcag actatctttc
     7801 tagggttaat ctagctgcat caggatcata tcgtcgggtc ttttttccgg ctcagtcatc
     7861 gcccaagctg gcgctatctg ggcatcgggg aggaagaagc ccgtgccttt tcccgcgagg
     7921 ttgaagcggc atggaaagag tttgccgagg atgactgctg ctgcattgac gttgagcgaa
     7981 aacgcacgtt taccatgatg attcgggaag gtgtggccat gcacgccttt aacggtgaac
     8041 tgttcgttca ggccacctgg gataccagtt cgtcgcggct tttccggaca cagttccgga
     8101 tggtcagccc gaagcgcatc agcaacccga acaataccgg cgacagccgg aactgccgtg
     8161 ccggtgtgca gattaatgac agcggtgcgg cgctgggata ttacgtcagc gaggacgggt
     8221 atcctggctg gatgccgcag aaatggacat ggataccccg tgagttaccc ggcgggcgcg
     8281 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     8341 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     8401 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     8461 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     8521 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     8581 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     8641 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     8701 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     8761 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     8821 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     8881 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     8941 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     9001 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     9061 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     9121 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     9181 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     9241 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     9301 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     9361 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     9421 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     9481 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     9541 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     9601 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     9661 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     9721 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     9781 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     9841 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     9901 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     9961 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
    10021 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
    10081 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
    10141 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
    10201 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
    10261 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcat
//
