LOCUS       sequence-205499-       11590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cutting NFS1 genomic locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11590)
  AUTHORS   Alvarez SW, Sviderskiy VO, Terzi EM, Papagiannakopoulos T, Moreira 
            AL, Adams S, Sabatini DM, Birsoy K, Possemato R
  TITLE     NFS1 undergoes positive selection in lung tumours and protects cells
            from ferroptosis.
  JOURNAL   Nature. 2017 Nov 22. doi: 10.1038/nature24637.
  PUBMED    29168506
REFERENCE   2  (bases 1 to 11590)
  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: "Nature. 
            2017 Nov 22. doi: 10.1038/nature24637."
COMMENT     Sequence Label: pLENTICRISPR sgNFS1
FEATURES             Location/Qualifiers
     source          1..11590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      44..632
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     533..552
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     786..803
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    920..941
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(956..961,962..979,980..986)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    994..1010
                     /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)."
     primer_bind     999..1021
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1018..1034
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1018..1034
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1053..1073
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1055..1073
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1101..1327
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1328..1508
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1555..1680
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2173..2406
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2591..2635
                     /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"
     promoter        2813..3061
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2813..2833
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2984..3003
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        3083..3158
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    3206..3323
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3392..3603
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3623..3632
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3632..3655
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3662..3682
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3707..7807
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             3707..7807
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7808..7855
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7871..7927
                     /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             7928..8527
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7928..7947)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8424..8444
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8543..9131
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8596..8616)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9014..9025)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9203..9436
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9514..9635
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9551..9570)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9605..9624
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9675..9810
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9830..9849)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9831..9849)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9859..9876)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9859..9875)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9868..9890)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10017..10472
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10104..10123)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10314..10335
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10498..10602
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10603..10671,10672..11463)
                     /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     complement(10821..10840)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
       61 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
      121 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
      181 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
      241 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
      301 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
      361 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
      421 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
      481 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
      541 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
      601 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
      661 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
      721 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
      781 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
      841 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
      901 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
      961 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     1021 gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa
     1081 agctggagct gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt gcaacatggt
     1141 aacgatgagt tagcaacatg ccttacaagg agagaaaaag caccgtgcat gccgattggt
     1201 ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa cagacgggtc tgacatggat
     1261 tggacgaacc actgaattgc cgcattgcag agatattgta tttaagtgcc tagctcgata
     1321 cataaacggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     1381 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     1441 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     1501 ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct
     1561 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg
     1621 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc
     1681 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga
     1741 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca
     1801 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa
     1861 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc
     1921 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag
     1981 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg
     2041 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat
     2101 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag
     2161 agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg
     2221 cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca
     2281 gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg
     2341 gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca
     2401 gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa
     2461 tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg
     2521 ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa
     2581 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa
     2641 ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg
     2701 cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg
     2761 atattcacca ttatcgtttc agacccacct cccaaccccg aggggaccca gagagggcct
     2821 atttcccatg attccttcat atttgcatat acgatacaag gctgttagag agataattag
     2881 aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata
     2941 atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac
     3001 cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac
     3061 cggcggattt gcagttccag aagttttaga gctagaaata gcaagttaaa ataaggctag
     3121 tccgttatca acttgaaaaa gtggcaccga gtcggtgctt ttttgaattc tagatcttga
     3181 gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg
     3241 caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac
     3301 aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat ccactttggc
     3361 gccggctcga gtggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     3421 gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta
     3481 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     3541 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     3601 caggtgtcgt gacgcgggat ccgccaccat ggattacaaa gacgatgacg ataagatggc
     3661 cccaaagaag aagcggaagg tcggtatcca cggagtccca gcagccgaca agaagtacag
     3721 catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa
     3781 ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa
     3841 cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag
     3901 aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt
     3961 cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag agtccttcct
     4021 ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt
     4081 ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac
     4141 cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg
     4201 ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat
     4261 ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt
     4321 ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat
     4381 cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct
     4441 gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct
     4501 gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta
     4561 cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct
     4621 gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga
     4681 cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa
     4741 gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg
     4801 agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac
     4861 cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga
     4921 caacggcagc atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca
     4981 ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt
     5041 ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac
     5101 cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc
     5161 ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa
     5221 ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa
     5281 agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa
     5341 ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga
     5401 ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg
     5461 gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt
     5521 cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt
     5581 tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa
     5641 agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct
     5701 gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga
     5761 cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga
     5821 ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa
     5881 tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga
     5941 gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga
     6001 gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga
     6061 gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct
     6121 gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca
     6181 ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt
     6241 tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa
     6301 gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct
     6361 gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg
     6421 cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca
     6481 gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa
     6541 tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt
     6601 ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga
     6661 cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag
     6721 cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga
     6781 gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt
     6841 caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa
     6901 cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt
     6961 gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag
     7021 caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg
     7081 ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt
     7141 ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat
     7201 caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg
     7261 ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct
     7321 ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact
     7381 ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa
     7441 gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct
     7501 ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa
     7561 tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc
     7621 cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta
     7681 ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac
     7741 cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg
     7801 aggcgacaag cgtcctgctg ctactaagaa agctggtcaa gctaagaaaa agaaagctag
     7861 cggcagcggc gccaccaact tcagcctgct gaagcaggcc ggcgacgtgg aggagaaccc
     7921 cggccccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
     7981 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     8041 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     8101 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     8161 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     8221 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     8281 caaggagccc gcgtggttcc tggccaccgt cggagtctcg cccgaccacc agggcaaggg
     8341 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     8401 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     8461 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     8521 tgcctgaacg cgttaagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     8581 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     8641 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     8701 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     8761 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     8821 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     8881 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     8941 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     9001 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     9061 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     9121 cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     9181 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg
     9241 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     9301 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     9361 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     9421 tggaaaatct ctagcagtac gtatagtagt tcatgtcatc ttattattca gtatttataa
     9481 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
     9541 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     9601 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
     9661 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     9721 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     9781 agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga
     9841 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     9901 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     9961 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc
    10021 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
    10081 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
    10141 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
    10201 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
    10261 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
    10321 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
    10381 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
    10441 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
    10501 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
    10561 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
    10621 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
    10681 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
    10741 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
    10801 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
    10861 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
    10921 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
    10981 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
    11041 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
    11101 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
    11161 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
    11221 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
    11281 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
    11341 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
    11401 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
    11461 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
    11521 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
    11581 gagttttcgt
//
