LOCUS       sequence-367766-       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 sgNRF2-3
FEATURES             Location/Qualifiers
     source          1..12824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(15..35)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        20..349
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      200..335
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     262..281
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        397..444
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             463..837
                     /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    967..1088
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1004..1023)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1058..1077
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1137..1153)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1137..1153)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1150..1172)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1161..1177
                     /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(1185..1191,1192..1209,1210..1215))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1230..1251
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1368..1385)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1539..2127)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1619..1638)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2298..3089,3090..3158))
                     /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     2921..2940
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3159..3263)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3338..3357)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3529..3908
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3910..4108
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4063..4083
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4126..4306
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4353..4478
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4971..5204
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5389..5433
                     /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    5731..5848
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5899..6147
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5899..5919
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6070..6089
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6169..6244
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        6306..6517
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6536..6545
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6542..10645
                     /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             6542..10645
                     /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             10646..10693
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             10694..10717
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             10727..10783
                     /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             10784..11182
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    11207..11795
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11260..11280)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11678..11689)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             11867..12100
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(12126..12143)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12132..12356
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(12402..12824,1..6)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12489..12508)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12699..12720
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
       61 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
      121 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
      181 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
      241 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
      301 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
      361 agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag
      421 tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt
      481 gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg
      541 ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg
      601 accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg
      661 tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc
      721 cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc
      781 gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac
      841 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
      901 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
      961 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     1021 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     1081 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     1141 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     1201 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     1261 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     1321 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     1381 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     1441 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     1501 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1561 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     1621 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     1681 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     1741 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     1801 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     1861 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     1921 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     1981 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2041 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2101 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2161 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     2221 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     2281 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     2341 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     2401 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     2461 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     2521 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     2581 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     2641 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     2701 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     2761 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     2821 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     2881 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     2941 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3001 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     3061 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3121 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     3181 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     3241 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     3301 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
     3361 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     3421 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
     3481 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat
     3541 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     3601 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     3661 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     3721 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     3781 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     3841 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     3901 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     3961 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     4021 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     4081 gtgtacggtg ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta
     4141 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     4201 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     4261 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca
     4321 gggacttgaa agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg
     4381 aagcgcgcac ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta
     4441 gcggaggcta gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta
     4501 gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac
     4561 atatagtatg ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa
     4621 catcagaagg ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag
     4681 aagaacttag atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag
     4741 agataaaaga caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga
     4801 ccaccgcaca gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat
     4861 tggagaagtg aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc
     4921 accaaggcaa agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg
     4981 ttccttgggt tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg
     5041 gtacaggcca gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct
     5101 attgaggcgc aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca
     5161 agaatcctgg ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc
     5221 tctggaaaac tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct
     5281 ctggaacaga tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac
     5341 acaagcttaa tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa
     5401 gaattattgg aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg
     5461 ctgtggtata taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt
     5521 tttgctgtac tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag
     5581 acccacctcc caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga
     5641 gagagagaca gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt
     5701 ctgcagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta
     5761 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
     5821 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt
     5881 ttggttaatt aaggtaccga gggcctattt cccatgattc cttcatattt gcatatacga
     5941 tacaaggctg ttagagagat aattagaatt aatttgactg taaacacaaa gatattagta
     6001 caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt
     6061 tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta
     6121 tatatcttgt ggaaaggacg aaacaccgtt acaactagat gaagagacgt tttagagcta
     6181 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     6241 gtgctttttt gaattcgcta gctaggtctt gaaaggagtg ggaattggct ccggtgcccg
     6301 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
     6361 ttgatccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
     6421 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
     6481 cgttcttttt cgcaacgggt ttgccgccag aacacaggac cggttctaga gcgctgccac
     6541 catggacaag aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt
     6601 gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg
     6661 gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga
     6721 ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg
     6781 ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag
     6841 actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg
     6901 caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa
     6961 gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca
     7021 catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga
     7081 cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc
     7141 catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag
     7201 acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa
     7261 cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga
     7321 ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc
     7381 ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat
     7441 cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc
     7501 tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg
     7561 gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc
     7621 cggctacatt gacggcggag ccagccagga agagttctac aagttcatca agcccatcct
     7681 ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg
     7741 gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca
     7801 cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat
     7861 cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag
     7921 cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga
     7981 agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa
     8041 gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt
     8101 gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct
     8161 gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac
     8221 cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat
     8281 ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat
     8341 tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt
     8401 gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc
     8461 ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg
     8521 caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct
     8581 ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga
     8641 cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct
     8701 gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac
     8761 agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt
     8821 gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag
     8881 aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc
     8941 cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg
     9001 ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca
     9061 tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag
     9121 cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa
     9181 gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct
     9241 gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca
     9301 gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa
     9361 cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc
     9421 caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa
     9481 ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa
     9541 gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa
     9601 gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag
     9661 caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg
     9721 gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt
     9781 tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt
     9841 gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat
     9901 cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc
     9961 ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt
    10021 gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga
    10081 ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa
    10141 gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact
    10201 gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag
    10261 ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga
    10321 acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt
    10381 gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa
    10441 gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc
    10501 ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa
    10561 agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat
    10621 cgacctgtct cagctgggag gcgacaagcg acctgccgcc acaaagaagg ctggacaggc
    10681 taagaagaag aaagattaca aagacgatga cgataaggga tccggcgcaa caaacttctc
    10741 tctgctgaaa caagccggag atgtcgaaga gaatcctgga ccgatggcca agcctttgtc
    10801 tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca tccccatctc
    10861 tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct tcactggtgt
    10921 caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg gcactgctgc
    10981 tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat
    11041 cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc
    11101 catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat tgctgccctc
    11161 tggttatgtg tgggagggct aagaattcga tatcaagctt atcggtaatc aacctctgga
    11221 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
    11281 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
    11341 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
    11401 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
    11461 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
    11521 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
    11581 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
    11641 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
    11701 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
    11761 gacgagtcgg atctcccttt gggccgcctc cccgcgtcga ctttaagacc aatgacttac
    11821 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
    11881 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
    11941 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
    12001 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
    12061 tccctcagac ccttttagtc agtgtggaaa atctctagca gggcccgttt aaacccgctg
    12121 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
    12181 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
    12241 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
    12301 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
    12361 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
    12421 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
    12481 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
    12541 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
    12601 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
    12661 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
    12721 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
    12781 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacg
//
