LOCUS       sequence-416685-       13214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual expression of Cas9 and sgRNA targeting ZNF746.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13214)
  AUTHORS   LaPak KM, Saeidi S, Bok I, Wamsley NT, Plutzer IB, Bhatt DP, Luo J, 
            Ashrafi G, Ben Major M
  TITLE     Proximity proteomic analysis of the NRF family reveals the 
            Parkinson's disease protein ZNF746/PARIS as a co-complexed repressor
            of NRF2.
  JOURNAL   Sci Signal. 2023 Dec 12;16(815):eadi9018. doi: 
            10.1126/scisignal.adi9018. Epub 2023 Dec 12.
  PUBMED    38085818
REFERENCE   2  (bases 1 to 13214)
  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: "Sci 
            Signal."; date: "2023-12-12"; volume: "16(815):eadi9018. doi"; 
            pages: " 10.1126/scisignal.adi9018. Epub 2023 Dec 12"
COMMENT     Sequence Label: pLenti-Cas9-ZNF746
FEATURES             Location/Qualifiers
     source          1..13214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             471..704
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(730..747)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    736..960
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1006..1434
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1093..1112)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1303..1324
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1443..1463)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1448..1777
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1628..1763
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1690..1709
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1825..1872
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1891..2265
                     /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    2395..2516
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2432..2451)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2486..2505
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2565..2581)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2565..2581)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2578..2600)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2589..2605
                     /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(2613..2619,2620..2637,2638..2643))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2658..2679
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2796..2813)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2967..3555)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3047..3066)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3726..4517,4518..4586))
                     /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     4349..4368
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4587..4691)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4766..4785)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4957..5336
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5338..5536
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5491..5511
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5554..5734
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5781..5906
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6399..6632
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6817..6861
                     /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    7159..7276
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7327..7575
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7327..7347
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7498..7517
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7597..7672
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        7734..7945
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      7964..7973
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7970..12073
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7970..12073
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             12074..12121
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             12122..12145
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             12155..12211
                     /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             12212..12610
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    12626..13214
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(12679..12699)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(13097..13108)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 gtcgacttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
       61 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
      121 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
      181 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
      241 gttgtgtgac tctggtaact agagatccac ytccctswtt krctcaactm aacvcarcgg
      301 ccmtkcctcm gckmgccmck gwccctgcgm tgskgcctct tccgcgtctt cgccttcgcc
      361 ctcagacgag tcggatctcc ctttgggccg cctccccgcg tcgactttaa gaccaatgac
      421 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
      481 aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga
      541 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
      601 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
      661 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagggccc gtttaaaccc
      721 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
      781 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
      841 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
      901 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
      961 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     1021 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     1081 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     1141 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     1201 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     1261 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     1321 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     1381 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     1441 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     1501 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     1561 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     1621 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     1681 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     1741 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     1801 cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata
     1861 gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg
     1921 ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc
     1981 cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc
     2041 agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc
     2101 ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc
     2161 gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac
     2221 ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt gctacgagat
     2281 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     2341 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     2401 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2461 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     2521 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     2581 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     2641 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     2701 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     2761 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2821 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2881 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2941 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3001 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3061 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3121 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3181 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3241 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3301 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3361 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3421 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3481 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3541 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3601 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3661 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3721 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3781 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3841 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3901 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3961 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4021 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4081 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4141 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4201 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4261 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4321 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4381 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4441 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4501 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4561 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4621 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4681 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     4741 ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     4801 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     4861 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
     4921 cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt
     4981 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     5041 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     5101 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     5161 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     5221 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     5281 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     5341 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     5401 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     5461 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     5521 aggtctatat aagcagcgcg ttttgcctgt actgggtctc tctggttaga ccagatctga
     5581 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     5641 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     5701 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
     5761 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
     5821 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
     5881 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
     5941 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
     6001 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
     6061 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     6121 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     6181 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     6241 aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa
     6301 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag
     6361 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc
     6421 ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga
     6481 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
     6541 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
     6601 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
     6661 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
     6721 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata
     6781 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
     6841 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
     6901 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
     6961 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
     7021 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga
     7081 gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat
     7141 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
     7201 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
     7261 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt ggttaattaa
     7321 ggtaccgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     7381 agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     7441 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     7501 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
     7561 aaaggacgaa acaccgggct ctacaagcac gtgatggttt tagagctaga aatagcaagt
     7621 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttga
     7681 attcgctagc taggtcttga aaggagtggg aattggctcc ggtgcccgtc agtgggcaga
     7741 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gatccggtgc
     7801 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
     7861 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
     7921 caacgggttt gccgccagaa cacaggaccg gttctagagc gctgccacca tggacaagaa
     7981 gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga
     8041 gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa
     8101 gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct
     8161 gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga
     8221 gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc
     8281 cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga
     8341 cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga
     8401 cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt
     8461 ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct
     8521 gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag
     8581 cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa
     8641 tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct
     8701 gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact
     8761 gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga
     8821 ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga
     8881 catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag
     8941 atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc
     9001 tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga
     9061 cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga
     9121 cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac
     9181 cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg
     9241 gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct
     9301 gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg
     9361 gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa
     9421 gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa
     9481 cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct
     9541 gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca
     9601 gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct
     9661 gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga
     9721 agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa
     9781 ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac
     9841 actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga
     9901 cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg
     9961 gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa
    10021 gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt
    10081 taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat
    10141 tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt
    10201 ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc
    10261 cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat
    10321 cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac
    10381 ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt
    10441 ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca
    10501 gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg
    10561 gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg
    10621 gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga
    10681 gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac
    10741 ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga
    10801 cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc
    10861 cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc
    10921 ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct
    10981 ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa
    11041 gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa
    11101 ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga
    11161 gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg
    11221 gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg
    11281 cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa
    11341 ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct
    11401 ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct
    11461 ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc
    11521 caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt
    11581 cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa
    11641 cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa
    11701 gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca
    11761 ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga
    11821 cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga
    11881 gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt
    11941 caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga
    12001 cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca
    12061 gctgggaggc gacaagcgac ctgccgccac aaagaaggct ggacaggcta agaagaagaa
    12121 agattacaaa gacgatgacg ataagggatc cggcgcaaca aacttctctc tgctgaaaca
    12181 agccggagat gtcgaagaga atcctggacc gatggccaag cctttgtctc aagaagaatc
    12241 caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag
    12301 cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca
    12361 ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc
    12421 tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg
    12481 cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga
    12541 cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg
    12601 ggagggctaa acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt
    12661 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
    12721 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
    12781 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
    12841 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
    12901 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
    12961 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
    13021 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
    13081 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
    13141 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
    13201 ggccgcctcc ccgc
//
