LOCUS       sequence-367906-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  knock out KEAP1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13013)
  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 13013)
  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 sgKEAP1-2
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..46)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(200..788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(280..299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(959..1750,1751..1819))
                     /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     1582..1601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1820..1924)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1999..2018)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2190..2569
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2571..2769
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2724..2744
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2787..2967
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3014..3139
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3632..3865
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4050..4094
                     /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    4392..4509
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4560..4808
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4560..4580
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4731..4750
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4830..4905
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        4967..5178
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5197..5206
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5203..9306
                     /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             5203..9306
                     /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             9307..9354
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9355..9378
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             9388..9444
                     /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             9445..10041
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     9938..9958
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    10057..10645
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10110..10130)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10528..10539)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10717..10950
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10976..10993)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10982..11206
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11252..11680
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11339..11358)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11549..11570
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11689..11709)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11694..12023
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11874..12009
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11936..11955
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12071..12118
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12137..12511
                     /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    12641..12762
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12678..12697)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12732..12751
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12811..12827)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12811..12827)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12824..12846)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12835..12851
                     /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(12859..12865,12866..12883,12884..12889))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12904..12925
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
       61 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
      121 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
      181 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
      241 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
      301 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
      361 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
      421 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
      481 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
      541 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
      601 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
      661 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
      721 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
      781 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
      841 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
      901 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
      961 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     1021 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     1081 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     1141 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     1201 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     1261 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     1321 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     1381 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     1441 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     1501 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     1561 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     1621 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     1681 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     1741 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     1801 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     1861 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     1921 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     1981 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     2041 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     2101 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
     2161 tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta
     2221 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     2281 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     2341 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     2401 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     2461 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     2521 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     2581 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     2641 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     2701 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     2761 tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg
     2821 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     2881 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     2941 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
     3001 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
     3061 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
     3121 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
     3181 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
     3241 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
     3301 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
     3361 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
     3421 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
     3481 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
     3541 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
     3601 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
     3661 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
     3721 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
     3781 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
     3841 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
     3901 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
     3961 acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta atacactcct
     4021 taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg gaattagata
     4081 aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat ataaaattat
     4141 tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta ctttctatag
     4201 tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc ccaaccccga
     4261 ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac agagacagat
     4321 ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca aatggcagta
     4381 ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata
     4441 gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt
     4501 caaaattttc gggtttatta cagggacagc agagatccag tttggttaat taaggtaccg
     4561 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga
     4621 taattagaat taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa
     4681 agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata
     4741 tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac
     4801 gaaacaccgt gtgtcctcca cgtcatgaag ttttagagct agaaatagca agttaaaata
     4861 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgaattcgct
     4921 agctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca
     4981 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgatccgg tgcctagaga
     5041 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     5101 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     5161 tttgccgcca gaacacagga ccggttctag agcgctgcca ccatggacaa gaagtacagc
     5221 atcggcctgg acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag
     5281 gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac
     5341 ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga
     5401 accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc
     5461 agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg
     5521 gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg
     5581 gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc
     5641 gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc
     5701 cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc
     5761 cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg
     5821 gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc
     5881 gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg
     5941 ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg
     6001 agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac
     6061 gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg
     6121 agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac
     6181 gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag
     6241 tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga
     6301 gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc
     6361 gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac
     6421 aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag
     6481 gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc
     6541 cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc
     6601 agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct
     6661 tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag
     6721 gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa
     6781 gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag
     6841 gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag
     6901 gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg
     6961 ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc
     7021 ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt
     7081 gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa
     7141 gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg
     7201 atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac
     7261 ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag
     7321 gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat
     7381 ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag
     7441 ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag
     7501 aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag
     7561 ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg
     7621 cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag
     7681 gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt
     7741 ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag
     7801 agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg
     7861 ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc
     7921 ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag
     7981 atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat
     8041 gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc
     8101 cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac
     8161 gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc
     8221 gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag
     8281 caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc
     8341 aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac
     8401 ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg
     8461 ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc
     8521 aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg
     8581 gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg
     8641 gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc
     8701 accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc
     8761 tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg
     8821 gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg
     8881 gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag
     8941 ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg
     9001 gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat
     9061 ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc
     9121 gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac
     9181 tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc
     9241 ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga
     9301 ggcgacaagc gacctgccgc cacaaagaag gctggacagg ctaagaagaa gaaagattac
     9361 aaagacgatg acgataaggg atccggcgca acaaacttct ctctgctgaa acaagccgga
     9421 gatgtcgaag agaatcctgg accgaccgag tacaagccca cggtgcgcct cgccacccgc
     9481 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     9541 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     9601 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     9661 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     9721 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     9781 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggagt ctcgcccgac
     9841 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     9901 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     9961 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
    10021 acccgcaagc ccggtgcctg aacgcgttaa gtcgacaatc aacctctgga ttacaaaatt
    10081 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
    10141 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
    10201 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
    10261 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
    10321 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
    10381 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
    10441 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
    10501 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
    10561 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
    10621 atctcccttt gggccgcctc cccgcgtcga ctttaagacc aatgacttac aaggcagctg
    10681 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
    10741 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
    10801 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
    10861 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
    10921 ccttttagtc agtgtggaaa atctctagca gggcccgttt aaacccgctg atcagcctcg
    10981 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
    11041 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
    11101 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
    11161 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
    11221 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
    11281 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
    11341 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
    11401 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
    11461 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
    11521 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
    11581 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
    11641 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
    11701 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
    11761 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
    11821 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
    11881 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
    11941 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
    12001 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
    12061 atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
    12121 ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga
    12181 cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga
    12241 ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga
    12301 ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta
    12361 cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac
    12421 cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg
    12481 cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc
    12541 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
    12601 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
    12661 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
    12721 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
    12781 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
    12841 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
    12901 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
    12961 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcg
//
