LOCUS       sequence-364866-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  knock out FSP1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13013)
  AUTHORS   Mao C, Liu X, Zhang Y, Lei G, Yan Y, Lee H, Koppula P, Wu S, Zhuang 
            L, Fang B, Poyurovsky MV, Olszewski K, Gan B
  TITLE     DHODH-mediated ferroptosis defence is a targetable vulnerability in 
            cancer.
  JOURNAL   Nature. 2021 May;593(7860):586-590. doi: 10.1038/s41586-021-03539-7.
            Epub 2021 May 12.
  PUBMED    33981038
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"; doi: 
            "10.1038/s41586-021-03539-7"; journalName: "Nature"; date: 
            "2021-05"; volume: "593"; issue: "7860"; pages: "586-590"
COMMENT     Sequence Label: lentiCRISPR v2-sgFSP1
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             6..186
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    233..358
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    851..1084
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1269..1313
                     /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    1611..1728
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1779..2027
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1779..1799
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1950..1969
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2049..2124
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2186..2397
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2416..2425
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2422..6525
                     /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             2422..6525
                     /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             6526..6573
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6574..6597
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6607..6663
                     /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             6664..7260
                     /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     7157..7177
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7276..7864
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7329..7349)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7747..7758)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7936..8169
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8195..8212)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8201..8425
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8471..8899
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8558..8577)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8768..8789
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8908..8928)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8913..9242
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9093..9228
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9155..9174
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9290..9337
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9356..9730
                     /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    9860..9981
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9897..9916)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9951..9970
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10030..10046)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10030..10046)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10043..10065)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10054..10070
                     /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(10078..10084,10085..10102,10103..10108))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10123..10144
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10261..10278)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10432..11020)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10512..10531)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11191..11982,11983..12051))
                     /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     11814..11833
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12052..12156)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12231..12250)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        12422..12801
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        12803..13001
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     12956..12976
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
       61 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
      121 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
      181 ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg
      241 acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag
      301 tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag
      361 tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa
      421 gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt
      481 taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc
      541 atcccttcag acaggatcag aagaacttag atcattatat aatacagtag caaccctcta
      601 ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga
      661 agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag
      721 gaggagatat gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg
      781 aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag
      841 cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg
      901 cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc
      961 agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg
     1021 gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc
     1081 tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg
     1141 ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg
     1201 acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc
     1261 agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt
     1321 ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct
     1381 tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat
     1441 attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag
     1501 gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat
     1561 cggcactgcg tgcgccaatt ctgcagacaa atggcagtat tcatccacaa ttttaaaaga
     1621 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     1681 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac
     1741 agggacagca gagatccagt ttggttaatt aaggtaccga gggcctattt cccatgattc
     1801 cttcatattt gcatatacga tacaaggctg ttagagagat aattagaatt aatttgactg
     1861 taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt
     1921 ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt
     1981 atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccgtc ccgattccac
     2041 cgagacctgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
     2101 gaaaaagtgg caccgagtcg gtgctttttt gaattcgcta gctaggtctt gaaaggagtg
     2161 ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     2221 ttggggggag gggtcggcaa ttgatccggt gcctagagaa ggtggcgcgg ggtaaactgg
     2281 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     2341 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggac
     2401 cggttctaga gcgctgccac catggacaag aagtacagca tcggcctgga catcggcacc
     2461 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
     2521 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc
     2581 gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc
     2641 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
     2701 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
     2761 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
     2821 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
     2881 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
     2941 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
     3001 cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct
     3061 gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag
     3121 aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
     3181 agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac
     3241 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
     3301 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
     3361 aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc
     3421 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
     3481 cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga agagttctac
     3541 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
     3601 aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag
     3661 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
     3721 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
     3781 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
     3841 accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag
     3901 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
     3961 ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga
     4021 atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc
     4081 aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag
     4141 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
     4201 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
     4261 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
     4321 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
     4381 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
     4441 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
     4501 atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg
     4561 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
     4621 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg
     4681 cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga
     4741 cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc
     4801 cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg
     4861 tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg
     4921 tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac
     4981 aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa
     5041 gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc
     5101 cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag
     5161 gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag
     5221 atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg
     5281 aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac
     5341 aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg
     5401 ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac
     5461 aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc
     5521 gccaagtact tcttctacag caacatcatg aactttttca agaccgagat taccctggcc
     5581 aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg
     5641 tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat
     5701 atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag
     5761 aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc
     5821 ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag
     5881 tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc
     5941 ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac
     6001 ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg
     6061 ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg
     6121 aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag
     6181 cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc
     6241 agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc
     6301 tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt
     6361 accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg
     6421 aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
     6481 ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacaagcg acctgccgcc
     6541 acaaagaagg ctggacaggc taagaagaag aaagattaca aagacgatga cgataaggga
     6601 tccggcgcaa caaacttctc tctgctgaaa caagccggag atgtcgaaga gaatcctgga
     6661 ccgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     6721 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     6781 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     6841 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     6901 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     6961 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     7021 cccgcgtggt tcctggccac cgtcggagtc tcgcccgacc accagggcaa gggtctgggc
     7081 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     7141 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     7201 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     7261 acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     7321 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     7381 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     7441 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     7501 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     7561 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     7621 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     7681 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     7741 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     7801 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     7861 ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     7921 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg
     7981 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     8041 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     8101 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     8161 tctctagcag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     8221 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     8281 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     8341 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     8401 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     8461 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     8521 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     8581 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     8641 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     8701 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     8761 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     8821 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     8881 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     8941 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     9001 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     9061 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     9121 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     9181 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     9241 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta
     9301 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc
     9361 caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt
     9421 ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt
     9481 ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac
     9541 cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt
     9601 gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg
     9661 ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga
     9721 gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     9781 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     9841 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     9901 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     9961 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
    10021 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
    10081 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
    10141 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
    10201 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
    10261 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
    10321 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
    10381 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
    10441 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
    10501 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
    10561 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
    10621 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
    10681 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
    10741 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
    10801 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
    10861 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
    10921 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
    10981 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
    11041 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
    11101 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
    11161 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
    11221 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
    11281 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
    11341 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
    11401 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
    11461 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
    11521 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
    11581 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
    11641 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
    11701 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
    11761 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
    11821 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
    11881 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
    11941 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
    12001 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
    12061 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
    12121 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
    12181 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
    12241 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
    12301 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
    12361 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
    12421 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
    12481 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
    12541 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
    12601 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
    12661 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
    12721 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
    12781 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
    12841 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
    12901 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
    12961 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cct
//
