LOCUS       sequence-365299-       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_FDX1-3
FEATURES             Location/Qualifiers
     source          1..11590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..253
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             254..434
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    481..606
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1099..1332
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1517..1561
                     /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        1739..1987
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1739..1759
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1910..1929
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2009..2084
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2132..2249
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2318..2529
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2549..2558
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2558..2581
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2588..2608
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2633..6733
                     /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             2633..6733
                     /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             6734..6781
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6797..6853
                     /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             6854..7453
                     /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(6854..6873)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7350..7370
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7469..8057
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7522..7542)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7940..7951)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8129..8362
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8440..8561
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8477..8496)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8531..8550
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8601..8736
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8756..8775)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8757..8775)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8785..8802)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8785..8801)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8794..8816)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8943..9398
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9030..9049)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9240..9261
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9424..9528
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9529..9597,9598..10389)
                     /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(9747..9766)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10560..11148
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11049..11068
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11302..11319
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11436..11457
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11472..11477,11478..11495,11496..11502)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11510..11526
                     /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     11515..11537
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11534..11550
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11534..11550
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11569..11589
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11571..11589
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aacaaaagct ggagctgcaa gcttaatgta gtcttatgca atactcttgt agtcttgcaa
       61 catggtaacg atgagttagc aacatgcctt acaaggagag aaaaagcacc gtgcatgccg
      121 attggtggaa gtaaggtggt acgatcgtgc cttattagga aggcaacaga cgggtctgac
      181 atggattgga cgaaccactg aattgccgca ttgcagagat attgtattta agtgcctagc
      241 tcgatacata aacgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
      301 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
      361 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
      421 gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag
      481 ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga
      541 ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag
      601 agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca
      661 gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga
      721 ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag
      781 ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca
      841 accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag
      901 atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag
      961 acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt
     1021 aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga
     1081 aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac
     1141 tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt
     1201 gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac
     1261 agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga
     1321 tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc
     1381 ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat
     1441 ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc
     1501 gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt
     1561 gtggaattgg tttaacataa caaattggct gtggtatata aaattattca taatgatagt
     1621 aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag
     1681 gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg gacccagaga
     1741 gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat
     1801 aattagaatt aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa
     1861 gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat
     1921 gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg
     1981 aaacaccgtg attctctgct agatgttggt tttagagcta gaaatagcaa gttaaaataa
     2041 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt gaattctaga
     2101 tcttgagaca aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt
     2161 acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac
     2221 aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccac
     2281 tttggcgccg gctcgagtgg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca
     2341 gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag aaggtggcgc
     2401 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga
     2461 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc
     2521 agaacacagg tgtcgtgacg cgggatccgc caccatggat tacaaagacg atgacgataa
     2581 gatggcccca aagaagaagc ggaaggtcgg tatccacgga gtcccagcag ccgacaagaa
     2641 gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga
     2701 gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa
     2761 gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct
     2821 gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga
     2881 gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc
     2941 cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga
     3001 cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga
     3061 cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt
     3121 ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct
     3181 gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag
     3241 cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa
     3301 tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct
     3361 gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact
     3421 gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga
     3481 ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga
     3541 catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag
     3601 atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc
     3661 tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga
     3721 cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga
     3781 cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac
     3841 cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg
     3901 gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct
     3961 gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg
     4021 gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa
     4081 gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa
     4141 cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct
     4201 gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca
     4261 gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct
     4321 gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga
     4381 agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa
     4441 ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac
     4501 actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga
     4561 cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg
     4621 gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa
     4681 gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt
     4741 taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat
     4801 tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt
     4861 ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc
     4921 cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat
     4981 cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac
     5041 ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt
     5101 ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca
     5161 gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg
     5221 gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg
     5281 gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga
     5341 gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac
     5401 ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga
     5461 cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc
     5521 cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc
     5581 ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct
     5641 ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa
     5701 gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa
     5761 ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga
     5821 gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg
     5881 gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg
     5941 cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa
     6001 ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct
     6061 ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct
     6121 ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc
     6181 caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt
     6241 cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa
     6301 cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa
     6361 gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca
     6421 ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga
     6481 cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga
     6541 gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt
     6601 caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga
     6661 cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca
     6721 gctgggaggc gacaagcgtc ctgctgctac taagaaagct ggtcaagcta agaaaaagaa
     6781 agctagcggc agcggcgcca ccaacttcag cctgctgaag caggccggcg acgtggagga
     6841 gaaccccggc cccatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     6901 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     6961 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     7021 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     7081 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     7141 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     7201 ccggcccaag gagcccgcgt ggttcctggc caccgtcgga gtctcgcccg accaccaggg
     7261 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     7321 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     7381 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     7441 gcccggtgcc tgaacgcgtt aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag
     7501 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     7561 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     7621 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     7681 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     7741 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     7801 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     7861 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     7921 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     7981 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     8041 ttgggccgcc tccccgcgtc gactttaaga ccaatgactt acaaggcagc tgtagatctt
     8101 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa
     8161 gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     8221 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     8281 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     8341 tcagtgtgga aaatctctag cagtacgtat agtagttcat gtcatcttat tattcagtat
     8401 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat
     8461 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     8521 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta
     8581 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     8641 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     8701 tattccagaa gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta
     8761 tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa
     8821 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     8881 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     8941 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     9001 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     9061 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     9121 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     9181 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     9241 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     9301 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     9361 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     9421 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     9481 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     9541 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     9601 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     9661 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     9721 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     9781 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     9841 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     9901 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     9961 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
    10021 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
    10081 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
    10141 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
    10201 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
    10261 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
    10321 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
    10381 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
    10441 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
    10501 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
    10561 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
    10621 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
    10681 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
    10741 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
    10801 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
    10861 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
    10921 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
    10981 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
    11041 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
    11101 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
    11161 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
    11221 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
    11281 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
    11341 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
    11401 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
    11461 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
    11521 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct
    11581 cactaaaggg
//
