LOCUS       sequence-364863-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  knock out DHODH 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-sgDHODH-2
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(117..137)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        122..451
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      302..437
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     364..383
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        499..546
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             565..939
                     /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    1069..1190
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1106..1125)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1160..1179
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1239..1255)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1239..1255)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1252..1274)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1263..1279
                     /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(1287..1293,1294..1311,1312..1317))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1332..1353
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1470..1487)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1641..2229)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1721..1740)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2400..3191,3192..3260))
                     /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     3023..3042
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3261..3365)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3440..3459)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3631..4010
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4012..4210
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4165..4185
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4228..4408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4455..4580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5073..5306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5491..5535
                     /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    5833..5950
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        6001..6249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     6001..6021
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6172..6191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6271..6346
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        6408..6619
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6638..6647
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6644..10747
                     /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             6644..10747
                     /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             10748..10795
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             10796..10819
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             10829..10885
                     /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             10886..11482
                     /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     11379..11399
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    11498..12086
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11551..11571)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11969..11980)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             12158..12391
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(12417..12434)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12423..12647
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(12693..13013,1..108)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12780..12799)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12990..13011
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
       61 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
      121 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
      181 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
      241 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
      301 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
      361 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
      421 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
      481 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
      541 tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc
      601 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
      661 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
      721 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac
      781 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg
      841 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc
      901 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat
      961 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     1021 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     1081 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     1141 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     1201 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     1261 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     1321 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     1381 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     1441 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     1501 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     1561 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     1621 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     1681 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1741 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1801 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1861 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1921 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1981 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2041 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2101 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2161 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     2221 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     2281 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     2341 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     2401 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2461 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     2521 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     2581 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     2641 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     2701 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     2761 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     2821 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     2881 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     2941 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3001 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3061 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     3121 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     3181 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     3241 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     3301 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     3361 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     3421 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     3481 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     3541 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     3601 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
     3661 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     3721 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     3781 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     3841 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     3901 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     3961 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     4021 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     4081 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     4141 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     4201 atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
     4261 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     4321 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     4381 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
     4441 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     4501 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     4561 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     4621 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     4681 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     4741 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     4801 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     4861 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     4921 ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat
     4981 aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga
     5041 gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg gttcttggga
     5101 gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc cagacaatta
     5161 ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc gcaacagcat
     5221 ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct ggctgtggaa
     5281 agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa actcatttgc
     5341 accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca gatttggaat
     5401 cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt aatacactcc
     5461 ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt ggaattagat
     5521 aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta tataaaatta
     5581 ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt actttctata
     5641 gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct cccaaccccg
     5701 aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga cagagacaga
     5761 tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac aaatggcagt
     5821 attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat
     5881 agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat
     5941 tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttaa ttaaggtacc
     6001 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
     6061 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
     6121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
     6181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
     6241 cgaaacaccg catcttataa agtccgtcca gttttagagc tagaaatagc aagttaaaat
     6301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgaattcgc
     6361 tagctaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac
     6421 atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgatccg gtgcctagag
     6481 aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga
     6541 gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg
     6601 gtttgccgcc agaacacagg accggttcta gagcgctgcc accatggaca agaagtacag
     6661 catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa
     6721 ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa
     6781 cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag
     6841 aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt
     6901 cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag agtccttcct
     6961 ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt
     7021 ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac
     7081 cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg
     7141 ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat
     7201 ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt
     7261 ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat
     7321 cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct
     7381 gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct
     7441 gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta
     7501 cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct
     7561 gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga
     7621 cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa
     7681 gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg
     7741 agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac
     7801 cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga
     7861 caacggcagc atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca
     7921 ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt
     7981 ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac
     8041 cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc
     8101 ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa
     8161 ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa
     8221 agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa
     8281 ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga
     8341 ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg
     8401 gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt
     8461 cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt
     8521 tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa
     8581 agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct
     8641 gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga
     8701 cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga
     8761 ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa
     8821 tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga
     8881 gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga
     8941 gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga
     9001 gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct
     9061 gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca
     9121 ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt
     9181 tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa
     9241 gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct
     9301 gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg
     9361 cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca
     9421 gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa
     9481 tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt
     9541 ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga
     9601 cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag
     9661 cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga
     9721 gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt
     9781 caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa
     9841 cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt
     9901 gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag
     9961 caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg
    10021 ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt
    10081 ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat
    10141 caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg
    10201 ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct
    10261 ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact
    10321 ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa
    10381 gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct
    10441 ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa
    10501 tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc
    10561 cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta
    10621 ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac
    10681 cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg
    10741 aggcgacaag cgacctgccg ccacaaagaa ggctggacag gctaagaaga agaaagatta
    10801 caaagacgat gacgataagg gatccggcgc aacaaacttc tctctgctga aacaagccgg
    10861 agatgtcgaa gagaatcctg gaccgaccga gtacaagccc acggtgcgcc tcgccacccg
    10921 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
    10981 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
    11041 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
    11101 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
    11161 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
    11221 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggag tctcgcccga
    11281 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
    11341 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
    11401 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
    11461 gacccgcaag cccggtgcct gaacgcgtta agtcgacaat caacctctgg attacaaaat
    11521 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
    11581 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
    11641 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
    11701 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
    11761 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
    11821 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
    11881 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
    11941 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
    12001 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
    12061 gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct
    12121 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
    12181 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
    12241 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
    12301 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
    12361 cccttttagt cagtgtggaa aatctctagc agggcccgtt taaacccgct gatcagcctc
    12421 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
    12481 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
    12541 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
    12601 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
    12661 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
    12721 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
    12781 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
    12841 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
    12901 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
    12961 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaa
//
