LOCUS       sequence-365000-       11590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus all in one CRISPR vector targeting human FDX1 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11590)
  AUTHORS   Tsvetkov P, Coy S, Petrova B, Dreishpoon M, Verma A, Abdusamad M, 
            Rossen J, Joesch-Cohen L, Humeidi R, Spangler RD, Eaton JK, Frenkel 
            E, Kocak M, Corsello SM, Lutsenko S, Kanarek N, Santagata S, Golub 
            TR
  TITLE     Copper induces cell death by targeting lipoylated TCA cycle 
            proteins.
  JOURNAL   Science. 2022 Mar 18;375(6586):1254-1261. doi: 
            10.1126/science.abf0529. Epub 2022 Mar 17.
  PUBMED    35298263
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"; doi: 
            "10.1126/science.abf0529"; journalName: "Science"; date: 
            "2022-03-18- 18"; volume: "375"; issue: "6586"; pages: "1254-1261"
COMMENT     Sequence Label: plentiCRISPv2_FDX1-1
FEATURES             Location/Qualifiers
     source          1..11590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     114..133
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     367..384
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    501..522
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(537..542,543..560,561..567)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    575..591
                     /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     580..602
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     599..615
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     599..615
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     634..654
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        636..654
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        682..908
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             909..1089
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1136..1261
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1754..1987
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2172..2216
                     /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        2394..2642
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2394..2414
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2565..2584
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2664..2739
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2787..2904
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2973..3184
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3204..3213
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3213..3236
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3243..3263
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3288..7388
                     /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             3288..7388
                     /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             7389..7436
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7452..7508
                     /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             7509..8108
                     /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(7509..7528)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8005..8025
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8124..8712
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8177..8197)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8595..8606)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8784..9017
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9095..9216
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9132..9151)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9186..9205
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9256..9391
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9411..9430)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9412..9430)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9440..9457)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9440..9456)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9449..9471)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9598..10053
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9685..9704)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9895..9916
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10079..10183
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10184..10252,10253..11044)
                     /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(10402..10421)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(11215..11590,1..213)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
       61 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
      121 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
      181 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
      241 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
      301 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
      361 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
      421 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
      481 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
      541 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
      601 ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa
      661 aagctggagc tgcaagctta atgtagtctt atgcaatact cttgtagtct tgcaacatgg
      721 taacgatgag ttagcaacat gccttacaag gagagaaaaa gcaccgtgca tgccgattgg
      781 tggaagtaag gtggtacgat cgtgccttat taggaaggca acagacgggt ctgacatgga
      841 ttggacgaac cactgaattg ccgcattgca gagatattgt atttaagtgc ctagctcgat
      901 acataaacgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
      961 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     1021 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     1081 tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc
     1141 tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt
     1201 gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt
     1261 cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg
     1321 aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc
     1381 agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca
     1441 accatccctt cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct
     1501 ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag acaagataga
     1561 ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg
     1621 gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa
     1681 ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa
     1741 gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg
     1801 gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc
     1861 agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct
     1921 ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac
     1981 agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga
     2041 atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt
     2101 gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa
     2161 accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga
     2221 attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag
     2281 gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg
     2341 gatattcacc attatcgttt cagacccacc tcccaacccc gaggggaccc agagagggcc
     2401 tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga gagataatta
     2461 gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat
     2521 aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta
     2581 ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca
     2641 ccggcaggcc gctggatcca gcggttttag agctagaaat agcaagttaa aataaggcta
     2701 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttgaatt ctagatcttg
     2761 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     2821 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     2881 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccactttgg
     2941 cgccggctcg agtggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
     3001 cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt
     3061 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
     3121 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
     3181 acaggtgtcg tgacgcggga tccgccacca tggattacaa agacgatgac gataagatgg
     3241 ccccaaagaa gaagcggaag gtcggtatcc acggagtccc agcagccgac aagaagtaca
     3301 gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca
     3361 aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga
     3421 acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga
     3481 gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg caagagatct
     3541 tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc
     3601 tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg
     3661 tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca
     3721 ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg
     3781 gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca
     3841 tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg
     3901 tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga
     3961 tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc
     4021 tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc
     4081 tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt
     4141 acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc
     4201 tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg
     4261 acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga
     4321 agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg
     4381 gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca
     4441 ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg
     4501 acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc
     4561 aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct
     4621 tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga
     4681 ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg
     4741 cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga
     4801 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca
     4861 aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa
     4921 aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag
     4981 aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc
     5041 ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact
     5101 tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt
     5161 ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca
     5221 aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc
     5281 tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg
     5341 acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag
     5401 aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca
     5461 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg
     5521 agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag
     5581 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag
     5641 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc
     5701 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc
     5761 aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct
     5821 ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca
     5881 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc
     5941 tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag
     6001 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc
     6061 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga
     6121 atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt
     6181 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg
     6241 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa
     6301 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg
     6361 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt
     6421 tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa
     6481 acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag
     6541 tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca
     6601 gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact
     6661 gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg
     6721 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga
     6781 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg
     6841 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc
     6901 tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac
     6961 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga
     7021 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc
     7081 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta
     7141 atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg
     7201 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt
     7261 actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca
     7321 ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg
     7381 gaggcgacaa gcgtcctgct gctactaaga aagctggtca agctaagaaa aagaaagcta
     7441 gcggcagcgg cgccaccaac ttcagcctgc tgaagcaggc cggcgacgtg gaggagaacc
     7501 ccggccccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca
     7561 gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg
     7621 atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg
     7681 ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca
     7741 cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt
     7801 tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc
     7861 ccaaggagcc cgcgtggttc ctggccaccg tcggagtctc gcccgaccac cagggcaagg
     7921 gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg
     7981 ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg
     8041 tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg
     8101 gtgcctgaac gcgttaagtc gacaatcaac ctctggatta caaaatttgt gaaagattga
     8161 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     8221 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     8281 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     8341 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     8401 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     8461 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     8521 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     8581 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     8641 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     8701 ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag atcttagcca
     8761 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct
     8821 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     8881 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     8941 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     9001 gtggaaaatc tctagcagta cgtatagtag ttcatgtcat cttattattc agtatttata
     9061 acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg
     9121 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     9181 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg
     9241 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     9301 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     9361 cagaagtagt gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg
     9421 agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     9481 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     9541 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
     9601 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     9661 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     9721 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     9781 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     9841 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     9901 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     9961 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
    10021 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
    10081 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
    10141 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
    10201 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
    10261 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
    10321 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
    10381 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
    10441 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
    10501 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
    10561 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
    10621 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
    10681 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
    10741 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
    10801 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
    10861 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
    10921 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
    10981 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
    11041 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
    11101 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
    11161 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
    11221 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
    11281 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
    11341 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
    11401 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
    11461 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
    11521 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
    11581 cgaactgaga
//
