LOCUS       sequence-367767-       12824 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  knock out NRF2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12824)
  AUTHORS   Koppula P, Lei G, Zhang Y, Yan Y, Mao C, Kondiparthi L, Shi J, Liu 
            X, Horbath A, Das M, Li W, Poyurovsky MV, Olszewski K, Gan B
  TITLE     A targetable CoQ-FSP1 axis drives ferroptosis- and 
            radiation-resistance in KEAP1 inactive lung cancers.
  JOURNAL   Nat Commun. 2022 Apr 22;13(1):2206. doi: 10.1038/s41467-022-29905-1.
  PUBMED    35459868
REFERENCE   2  (bases 1 to 12824)
  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: "Nat 
            Commun."; date: "2022-04-22"; volume: "13(1):2206. doi"; pages: " 
            10.1038/s41467-022-29905-1"
COMMENT     Sequence Label: lentiCRISPR v2 sgNRF2-4
FEATURES             Location/Qualifiers
     source          1..12824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     25..46
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(165..185)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        170..499
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      350..485
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     412..431
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        547..594
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             613..987
                     /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    1117..1238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1154..1173)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1208..1227
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1287..1303)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1287..1303)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1300..1322)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1311..1327
                     /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(1335..1341,1342..1359,1360..1365))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1380..1401
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1518..1535)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1689..2277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1769..1788)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2448..3239,3240..3308))
                     /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     3071..3090
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3309..3413)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3488..3507)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3679..4058
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4060..4258
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4213..4233
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4276..4456
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4503..4628
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5121..5354
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5539..5583
                     /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    5881..5998
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6049..6297
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6049..6069
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6220..6239
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6319..6394
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        6456..6667
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6686..6695
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6692..10795
                     /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             6692..10795
                     /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             10796..10843
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             10844..10867
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             10877..10933
                     /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             10934..11332
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    11357..11945
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11410..11430)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11828..11839)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             12017..12250
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(12276..12293)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12282..12506
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(12552..12824,1..156)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12639..12658)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
       61 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
      121 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
      181 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
      241 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
      301 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
      361 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
      421 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
      481 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
      541 gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
      601 aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc
      661 gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac
      721 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag
      781 gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
      841 gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag
      901 atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg
      961 cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc
     1021 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     1081 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     1141 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     1201 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     1261 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1321 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1381 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1441 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1501 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1561 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1621 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1681 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1741 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     1801 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     1861 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     1921 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     1981 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2041 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2101 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2161 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2221 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2281 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2341 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2401 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2461 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2521 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2581 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2641 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     2701 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     2761 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     2821 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     2881 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     2941 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3001 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3061 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3121 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3181 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3241 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3301 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3361 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3421 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     3481 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     3541 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     3601 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     3661 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     3721 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     3781 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     3841 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     3901 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     3961 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     4021 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     4081 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     4141 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     4201 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg
     4261 cgttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     4321 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     4381 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     4441 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
     4501 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
     4561 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
     4621 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
     4681 tagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac
     4741 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac
     4801 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag
     4861 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca
     4921 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc
     4981 agacctggag gaggagatat gagggacaat tggagaagtg aattatataa atataaagta
     5041 gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga
     5101 gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc
     5161 actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata
     5221 gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc
     5281 acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag
     5341 gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg
     5401 ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg
     5461 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa
     5521 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt
     5581 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata
     5641 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt
     5701 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac
     5761 aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta
     5821 gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa atggcagtat tcatccacaa
     5881 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat
     5941 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg
     6001 ggtttattac agggacagca gagatccagt ttggttaatt aaggtaccga gggcctattt
     6061 cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat aattagaatt
     6121 aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt
     6181 cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta
     6241 acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccgca
     6301 gatccactgg tttctgacgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg
     6361 ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt gaattcgcta gctaggtctt
     6421 gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     6481 ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa ggtggcgcgg
     6541 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     6601 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     6661 aacacaggac cggttctaga gcgctgccac catggacaag aagtacagca tcggcctgga
     6721 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
     6781 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc
     6841 cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag
     6901 aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat
     6961 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
     7021 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
     7081 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
     7141 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
     7201 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
     7261 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
     7321 catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc
     7381 cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc
     7441 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
     7501 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
     7561 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
     7621 cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca
     7681 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
     7741 tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga
     7801 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
     7861 cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat
     7921 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta
     7981 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
     8041 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
     8101 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag
     8161 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
     8221 gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt
     8281 gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga
     8341 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
     8401 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
     8461 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
     8521 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
     8581 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
     8641 gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
     8701 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
     8761 cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa
     8821 agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag
     8881 ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt
     8941 gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac
     9001 ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga
     9061 gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa
     9121 gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat
     9181 caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga
     9241 ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt
     9301 gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa
     9361 gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga
     9421 actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca
     9481 cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat
     9541 ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt
     9601 ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa
     9661 cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta
     9721 cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg
     9781 caaggctacc gccaagtact tcttctacag caacatcatg aactttttca agaccgagat
     9841 taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg
     9901 ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc
     9961 ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat
    10021 cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa
    10081 gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga
    10141 aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga
    10201 aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt
    10261 gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg
    10321 gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc
    10381 caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga
    10441 ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat
    10501 cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt
    10561 gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat
    10621 ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac
    10681 catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca
    10741 gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacaagcg
    10801 acctgccgcc acaaagaagg ctggacaggc taagaagaag aaagattaca aagacgatga
    10861 cgataaggga tccggcgcaa caaacttctc tctgctgaaa caagccggag atgtcgaaga
    10921 gaatcctgga ccgatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc
    10981 aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct
    11041 ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg
    11101 tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat
    11161 cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt
    11221 gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg gacagccgac
    11281 ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct aagaattcga
    11341 tatcaagctt atcggtaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
    11401 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
    11461 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
    11521 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
    11581 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
    11641 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
    11701 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
    11761 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
    11821 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
    11881 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
    11941 cccgcgtcga ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
    12001 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt
    12061 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
    12121 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
    12181 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
    12241 atctctagca gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
    12301 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
    12361 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
    12421 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
    12481 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
    12541 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
    12601 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
    12661 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
    12721 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
    12781 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttga
//
