LOCUS       sequence-424951-       11790 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  inducible expression of TDP-43 harboring mutant NLS and C-terminal 
            mClover3. Expression yields cytosolic diffuse TDP-43.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11790)
  AUTHORS   Sweeney KM, Chantarawong S, Barbieri EM, Cajka G, Liu M, Spruce L, 
            Fazelinia H, Portz B, Copley K, Lapidot T, Duhamel L, Greenwald P, 
            Saida N, Shalgi R, Shorter J, Shalem O
  TITLE     CRISPR screen for protein inclusion formation uncovers a role for 
            SRRD in the regulation of intermediate filament dynamics and 
            aggresome assembly.
  JOURNAL   PLoS Genet. 2024 Feb 5;20(2):e1011138. doi: 
            10.1371/journal.pgen.1011138. eCollection 2024 Feb.
  PUBMED    38315730
REFERENCE   2  (bases 1 to 11790)
  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: "PLoS 
            Genet."; date: "2024-02-5"; volume: "20(2):e1011138. doi"; pages: " 
            10.1371/journal.pgen.1011138. eCollection 2024 Feb"
COMMENT     Sequence Label: TRE-TDP-43-Delta-NLS-mClover3
FEATURES             Location/Qualifiers
     source          1..11790
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     55..76
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(195..215)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        200..529
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      380..515
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     442..461
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        577..624
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             643..1017
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    1147..1268
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1184..1203)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1238..1257
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1317..1333)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1317..1333)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1330..1352)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1341..1357
                     /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(1365..1371,1372..1389,1390..1395))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1410..1431
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1548..1565)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1719..2307)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1799..1818)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2478..3269,3270..3338))
                     /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     3101..3120
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3339..3443)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3518..3537)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3709..4088
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4090..4288
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             4306..4486
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4533..4658
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5151..5384
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5569..5613
                     /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    5911..6028
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6123..6437
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    6131..6149
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6167..6185
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6202..6220
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6238..6256
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6274..6292
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6309..6327
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6345..6363
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     6410..6434
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(7721..7723,7724..8434)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(7763..7784)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     8371..8392
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             8444..8500
                     /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             8501..9301
                     /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="GSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(8561..8580)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9171..9190
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     promoter        9357..9568
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             join(9593..10210,10211..10216,10217..10252,10253..10255,
                     10256..10291,10292..10294,10295..10330,10331..10339)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(10379..10399)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10797..10808)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             11013..11246
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(11272..11289)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11278..11502
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(11548..11790,1..186)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11635..11654)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
       61 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
      121 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
      181 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
      241 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
      301 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
      361 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
      421 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
      481 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
      541 agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag
      601 tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt
      661 gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg
      721 ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg
      781 accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg
      841 tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc
      901 cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc
      961 gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac
     1021 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     1081 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     1141 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     1201 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     1261 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     1321 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     1381 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     1441 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     1501 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     1561 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     1621 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     1681 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1741 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     1801 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     1861 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     1921 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     1981 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2041 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2101 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     2161 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2221 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2281 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2341 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     2401 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     2461 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     2521 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     2581 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     2641 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     2701 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     2761 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     2821 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     2881 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     2941 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3001 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3061 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3121 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3181 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     3241 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3301 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     3361 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     3421 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     3481 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
     3541 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     3601 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
     3661 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat
     3721 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     3781 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     3841 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     3901 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     3961 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     4021 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     4081 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     4141 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     4201 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     4261 gtgtacggtg ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta
     4321 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     4381 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     4441 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     4501 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     4561 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     4621 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     4681 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     4741 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     4801 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     4861 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     4921 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     4981 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     5041 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     5101 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     5161 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     5221 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     5281 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     5341 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     5401 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     5461 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     5521 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     5581 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     5641 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     5701 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     5761 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
     5821 gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt
     5881 ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta
     5941 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
     6001 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt
     6061 ttggttaatc gggtttatta cagggacagc agagatccac tttggccgcg aatcgatatg
     6121 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
     6181 agagaacgat gtcgagttta ctccctatca gtgatagaga acgtatgtcg agtttactcc
     6241 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
     6301 cgagtttatc cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag
     6361 agaacgtatg tcgaggtagg cgtgtacggt gggaggccta tataagcaga gctcgtttag
     6421 tgaaccgtca gatcgcctgg agaattggca catgtctgaa tatattcggg taaccgaaga
     6481 tgagaacgat gagcccattg aaataccatc ggaagacgat gggacggtgc tgctctccac
     6541 ggttacagcc cagtttccag gggcgtgtgg gcttcgctac aggaatccag tgtctcagtg
     6601 tatgagaggt gtccggctgg tagaaggaat tctgcatgcc ccagatgctg gctggggaaa
     6661 tctggtgtat gttgtcaact atccaaaaga taacgcagca gcaatggatg agacagatgc
     6721 ttcatcagca gtggcagtgg cagcagcagc agtccagaaa acatccgatt taatagtgtt
     6781 gggtctccca tggaaaacaa ccgaacagga cctgaaagag tattttagta cctttggaga
     6841 agttcttatg gtgcaggtca agaaagatct taagactggt cattcaaagg ggtttggctt
     6901 tgttcgtttt acggaatatg aaacacaagt gaaagtaatg tcacagcgac atatgataga
     6961 tggacgatgg tgtgactgca aacttcctaa ttctaagcaa agccaagatg agcctttgag
     7021 aagcagaaaa gtgtttgtgg ggcgctgtac agaggacatg actgaggatg agctgcggga
     7081 gttcttctct cagtacgggg atgtgatgga tgtcttcatc cccaagccat tcagggcctt
     7141 tgcctttgtt acatttgcag atgatcagat tgcgcagtct ctttgtggag aggacttgat
     7201 cattaaagga atcagcgttc atatatccaa tgccgaacct aagcacaata gcaatagaca
     7261 gttagaaaga agtggaagat ttggtggtaa tccaggtggc tttgggaatc agggtggatt
     7321 tggtaatagc agagggggtg gagctggttt gggaaacaat caaggtagta atatgggtgg
     7381 tgggatgaac tttggtgcgt tcagcattaa tccagccatg atggctgccg cccaggcagc
     7441 actacagagc agttggggta tgatgggcat gttagccagc cagcagaacc agtcaggccc
     7501 atcgggtaat aaccaaaacc aaggcaacat gcagagggag ccaaaccagg ccttcggttc
     7561 tggaaataac tcttatagtg gctctaattc tggtgcagca attggttggg gatcaacatc
     7621 caatgcaggg tcgggcagtg gttttaatgg aggctttggc tcaagcatgg attctaagtc
     7681 ttctggctgg ggaatgggat ccggaagtgg ctcaagcgga gtgagcaagg gcgaggagct
     7741 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     7801 cagcgtccgc ggcgagggcg agggcgatgc caccaacggc aagctgaccc tgaagttcat
     7861 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg
     7921 cgtggcctgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     7981 catgcccgaa ggctacgtcc aggagcgcac catctctttc aaggacgacg gtacctacaa
     8041 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     8101 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca acttcaacag
     8161 ccactacgtc tatatcacgg ccgacaagca gaagaactgc atcaaggcta acttcaagat
     8221 ccgccacaac gttgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     8281 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagccatc agtccaagct
     8341 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     8401 cgggattaca catggcatgg acgagctgta caagggatcc ggcgcaacaa acttctctct
     8461 gctgaaacaa gccggagatg tcgaagagaa tcctggaccg ggatcggcca ttgaacaaga
     8521 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     8581 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     8641 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     8701 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     8761 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     8821 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     8881 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     8941 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     9001 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     9061 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     9121 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     9181 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     9241 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     9301 agaattcgct agctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagtgggc
     9361 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgatccgg
     9421 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     9481 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     9541 tcgcaacggg tttgccgcca gaacacagga ccggttctag agcgctgcca ccatgtctag
     9601 actggacaag agcaaagtca taaacggagc tctggaatta ctcaatggtg tcggtatcga
     9661 aggcctgacg acaaggaaac tcgctcaaaa gctgggagtt gagcagccta ccctgtactg
     9721 gcacgtgaag aacaagcggg ccctgctcga tgccctgcca atcgagatgc tggacaggca
     9781 tcatacccac ttctgccccc tggaaggcga gtcatggcaa gactttctgc ggaacaacgc
     9841 caagtcatac cgctgtgctc tcctctcaca tcgcgacggg gctaaagtgc atctcggcac
     9901 ccgcccaaca gagaaacagt acgaaaccct ggaaaatcag ctcgcgttcc tgtgtcagca
     9961 aggcttctcc ctggagaacg cactgtacgc tctgtccgcc gtgggccact ttacactggg
    10021 ctgcgtattg gaggaacagg agcatcaagt agcaaaagag gaaagagaga cacctaccac
    10081 cgattctatg cccccacttc tgagacaagc aattgagctg ttcgaccggc agggagccga
    10141 acctgccttc cttttcggcc tggaactaat catatgtggc ctggagaaac agctaaagtg
    10201 cgaaagcggc gggccgaccg acgcccttga cgattttgac ttagacatgc tcccagccga
    10261 tgcccttgac gactttgacc ttgatatgct gcctgctgac gctcttgacg attttgacct
    10321 tgacatgctc cccgggtaac taagtaagga tcgatccaag atatcgtatt cttaactatg
    10381 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
    10441 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
    10501 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
    10561 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
    10621 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
    10681 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
    10741 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
    10801 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
    10861 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctgttt
    10921 cgcctcggcg tccggactag aggtaccttt aagaccaatg acttacaagg cagctgtaga
    10981 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
    11041 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
    11101 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
    11161 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
    11221 ttagtcagtg tggaaaatct ctagcagggc ccgtttaaac ccgctgatca gcctcgactg
    11281 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
    11341 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
    11401 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
    11461 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
    11521 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
    11581 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
    11641 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
    11701 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
    11761 attagggtga tggttcacgt agtgggccat
//
