LOCUS       sequence-416687-       13218 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Dual expression of Cas9 and non-specific sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13218)
  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 13218)
  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-NS-B
FEATURES             Location/Qualifiers
     source          1..13218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             475..708
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(734..751)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    740..964
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1010..1438
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1097..1116)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1307..1328
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1447..1467)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1452..1781
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1632..1767
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1694..1713
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1829..1876
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1895..2269
                     /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    2399..2520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2436..2455)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2490..2509
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2569..2585)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2569..2585)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2582..2604)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2593..2609
                     /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(2617..2623,2624..2641,2642..2647))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2662..2683
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2800..2817)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2971..3559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3051..3070)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3730..4521,4522..4590))
                     /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     4353..4372
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4591..4695)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4770..4789)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4961..5340
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5342..5540
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5495..5515
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5558..5738
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5785..5910
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6403..6636
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6821..6865
                     /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    7163..7280
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7331..7579
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7331..7351
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7502..7521
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7601..7676
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        7738..7949
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      7968..7977
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7974..12077
                     /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             7974..12077
                     /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             12078..12125
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             12126..12149
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             12159..12215
                     /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             12216..12614
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    12630..13218
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(12683..12703)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(13101..13112)
                     /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 caagtagkkg kkgccrwswk
      241 wtsygtgayt ctggtaacta gagatccctc rgctacttcc ctnmttsggm cctcaatcca
      301 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
      361 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa
      421 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
      481 ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt
      541 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      601 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      661 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa
      721 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
      781 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
      841 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
      901 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
      961 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     1021 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     1081 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     1141 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     1201 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     1261 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     1321 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     1381 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     1441 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     1501 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     1561 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     1621 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     1681 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     1741 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     1801 tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg
     1861 catagtataa tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc
     1921 ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt
     1981 ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt
     2041 catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg
     2101 cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc
     2161 ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg
     2221 cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg
     2281 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     2341 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     2401 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     2461 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     2521 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     2581 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     2641 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2701 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2761 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2821 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2881 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2941 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3001 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3061 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3121 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3181 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3241 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3301 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3361 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3421 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3481 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3541 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3601 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3661 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3721 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3781 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3841 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3901 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3961 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     4021 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     4081 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     4141 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     4201 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     4261 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     4321 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     4381 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4441 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4501 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4561 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4621 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4681 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     4741 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4801 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     4861 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
     4921 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
     4981 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     5041 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     5101 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     5161 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     5221 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     5281 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     5341 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     5401 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     5461 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     5521 tgggaggtct atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat
     5581 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     5641 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     5701 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
     5761 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
     5821 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
     5881 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
     5941 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
     6001 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
     6061 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
     6121 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
     6181 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
     6241 cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     6301 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     6361 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     6421 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     6481 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     6541 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     6601 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     6661 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     6721 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     6781 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     6841 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     6901 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     6961 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     7021 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     7081 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
     7141 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     7201 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     7261 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttaa
     7321 ttaaggtacc gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc
     7381 tgttagagag ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac
     7441 gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat
     7501 ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt catatatctt
     7561 gtggaaagga cgaaacaccg gtcatcaagg agcattccgt gttttagagc tagaaatagc
     7621 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     7681 ttgaattcgc tagctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg
     7741 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg
     7801 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     7861 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     7921 ttcgcaacgg gtttgccgcc agaacacagg accggttcta gagcgctgcc accatggaca
     7981 agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg
     8041 acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca
     8101 tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc
     8161 ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc
     8221 aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag
     8281 agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg
     8341 tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg
     8401 tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca
     8461 agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca
     8521 agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg
     8581 ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg
     8641 aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg
     8701 ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca
     8761 aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg
     8821 gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga
     8881 gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca
     8941 agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc
     9001 tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca
     9061 ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga
     9121 tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc
     9181 ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg cacgccattc
     9241 tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga
     9301 tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg
     9361 cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg
     9421 acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc
     9481 ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg
     9541 agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg
     9601 agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc
     9661 agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg
     9721 tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg
     9781 acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc
     9841 tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt
     9901 tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga
     9961 gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc
    10021 tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga
    10081 cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc
    10141 acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg
    10201 tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa
    10261 tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc
    10321 ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa
    10381 acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt
    10441 acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc
    10501 ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga
    10561 accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact
    10621 ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg
    10681 ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg
    10741 aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt
    10801 acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg
    10861 tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc
    10921 acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta
    10981 agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg
    11041 ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca
    11101 tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga
    11161 tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg
    11221 tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag
    11281 gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa
    11341 agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg
    11401 tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc
    11461 tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg
    11521 aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc
    11581 tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg
    11641 gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg
    11701 agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca
    11761 agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg
    11821 ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca
    11881 gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg
    11941 ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc
    12001 tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt
    12061 ctcagctggg aggcgacaag cgacctgccg ccacaaagaa ggctggacag gctaagaaga
    12121 agaaagatta caaagacgat gacgataagg gatccggcgc aacaaacttc tctctgctga
    12181 aacaagccgg agatgtcgaa gagaatcctg gaccgatggc caagcctttg tctcaagaag
    12241 aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact
    12301 acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat
    12361 atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg
    12421 cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc
    12481 cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga
    12541 aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg
    12601 tgtgggaggg ctaaacgcgt taagtcgaca atcaacctct ggattacaaa atttgtgaaa
    12661 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
    12721 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
    12781 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
    12841 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
    12901 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
    12961 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
    13021 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
    13081 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
    13141 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
    13201 tttgggccgc ctccccgc
//
