LOCUS       sequence-365312-       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_FDX1-2
FEATURES             Location/Qualifiers
     source          1..11590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      32..167
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(187..206)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(188..206)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(216..233)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(216..232)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(225..247)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      374..829
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(461..480)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     671..692
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        855..959
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(960..1028,1029..1820)
                     /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(1178..1197)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1991..2579
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2480..2499
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2733..2750
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2867..2888
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2903..2908,2909..2926,2927..2933)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2941..2957
                     /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     2946..2968
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2965..2981
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2965..2981
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3000..3020
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3002..3020
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3048..3274
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             3275..3455
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3502..3627
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4120..4353
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4538..4582
                     /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        4760..5008
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4760..4780
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4931..4950
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        5030..5105
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    5153..5270
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5339..5550
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5570..5579
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5579..5602
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             5609..5629
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5654..9754
                     /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             5654..9754
                     /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             9755..9802
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9818..9874
                     /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             9875..10474
                     /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(9875..9894)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     10371..10391
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    10490..11078
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10543..10563)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10961..10972)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             11150..11383
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    11461..11582
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11498..11517)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11552..11571
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
       61 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
      121 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc
      181 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
      241 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
      301 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
      361 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
      421 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
      481 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
      541 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
      601 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
      661 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
      721 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
      781 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
      841 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
      901 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
      961 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     1021 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     1081 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     1141 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     1201 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     1261 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     1321 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     1381 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     1441 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     1501 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     1561 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     1621 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     1681 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     1741 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     1801 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     1861 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     1921 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     1981 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2041 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2101 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     2161 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2221 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     2281 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     2341 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     2401 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     2461 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2521 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     2581 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     2641 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     2701 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     2761 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     2821 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     2881 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     2941 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
     3001 caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc
     3061 aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga
     3121 gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg
     3181 aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga
     3241 tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg
     3301 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     3361 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     3421 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
     3481 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
     3541 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
     3601 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
     3661 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
     3721 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
     3781 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
     3841 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
     3901 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
     3961 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     4021 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     4081 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     4141 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     4201 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     4261 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     4321 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     4381 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     4441 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     4501 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     4561 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     4621 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     4681 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     4741 aaccccgagg ggacccagag agggcctatt tcccatgatt ccttcatatt tgcatatacg
     4801 atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa agatattagt
     4861 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
     4921 ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt
     4981 atatatcttg tggaaaggac gaaacaccga gcggcctgct gaggaaccgg ttttagagct
     5041 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
     5101 ggtgcttttt tgaattctag atcttgagac aaatggcagt attcatccac aattttaaaa
     5161 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
     5221 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
     5281 acagggacag cagagatcca ctttggcgcc ggctcgagtg gctccggtgc ccgtcagtgg
     5341 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc
     5401 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     5461 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     5521 tttcgcaacg ggtttgccgc cagaacacag gtgtcgtgac gcgggatccg ccaccatgga
     5581 ttacaaagac gatgacgata agatggcccc aaagaagaag cggaaggtcg gtatccacgg
     5641 agtcccagca gccgacaaga agtacagcat cggcctggac atcggcacca actctgtggg
     5701 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
     5761 caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga
     5821 aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa
     5881 ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt
     5941 cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc
     6001 catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca
     6061 cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc
     6121 cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga
     6181 caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga
     6241 ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag
     6301 caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct
     6361 gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga
     6421 cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa
     6481 cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc
     6541 cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct
     6601 gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc
     6661 tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa
     6721 cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa
     6781 gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga
     6841 cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg
     6901 agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg
     6961 ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag
     7021 gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa
     7081 cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa
     7141 cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta
     7201 cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc
     7261 cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg
     7321 gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc
     7381 cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct
     7441 gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga
     7501 agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa
     7561 aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac
     7621 cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa
     7681 gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat
     7741 ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg
     7801 cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat
     7861 cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga
     7921 gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag
     7981 ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa
     8041 agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca
     8101 gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga
     8161 tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct
     8221 gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa
     8281 gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt
     8341 cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat
     8401 caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc
     8461 ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac
     8521 cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga
     8581 gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct
     8641 gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga
     8701 cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt
     8761 cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat
     8821 ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg
     8881 ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa
     8941 gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga
     9001 taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc
     9061 caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact
     9121 gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa
     9181 tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa
     9241 gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc
     9301 cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta
     9361 cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct
     9421 gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc
     9481 caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca
     9541 ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa
     9601 tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac
     9661 cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga
     9721 gacacggatc gacctgtctc agctgggagg cgacaagcgt cctgctgcta ctaagaaagc
     9781 tggtcaagct aagaaaaaga aagctagcgg cagcggcgcc accaacttca gcctgctgaa
     9841 gcaggccggc gacgtggagg agaaccccgg ccccatgacc gagtacaagc ccacggtgcg
     9901 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     9961 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
    10021 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
    10081 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
    10141 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
    10201 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
    10261 agtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
    10321 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
    10381 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
    10441 cacctggtgc atgacccgca agcccggtgc ctgaacgcgt taagtcgaca atcaacctct
    10501 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
    10561 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
    10621 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
    10681 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
    10741 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
    10801 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
    10861 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
    10921 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
    10981 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
    11041 tcagacgagt cggatctccc tttgggccgc ctccccgcgt cgactttaag accaatgact
    11101 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
    11161 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
    11221 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
    11281 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
    11341 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtacgta tagtagttca
    11401 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
    11461 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
    11521 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
    11581 tatcatgtct
//
