LOCUS       sequence-349894-       11590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral vector expressing Cas9 and a sgRNA targeting SDHB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11590)
  AUTHORS   Spinelli JB, Rosen PC, Sprenger HG, Puszynska AM, Mann JL, Roessler 
            JM, Cangelosi AL, Henne A, Condon KJ, Zhang T, Kunchok T, Lewis CA, 
            Chandel NS, Sabatini DM
  TITLE     Fumarate is a terminal electron acceptor in the mammalian electron 
            transport chain.
  JOURNAL   Science. 2021 Dec 3;374(6572):1227-1237. doi: 
            10.1126/science.abi7495. Epub 2021 Dec 2.
  PUBMED    34855504
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.abi7495"; journalName: "Science"; date: "2021-12-3-
            3"; volume: "374"; issue: "6572"; pages: "1227-1237"
COMMENT     Sequence Label: pLentiCRISPRv1_SDHB
FEATURES             Location/Qualifiers
     source          1..11590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    123..144
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(159..164,165..182,183..189)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    197..213
                     /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     202..224
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     221..237
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     221..237
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     256..276
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        258..276
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        304..530
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             531..711
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    758..883
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1376..1609
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1794..1838
                     /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        2016..2264
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2016..2036
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2187..2206
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2286..2361
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2409..2526
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2595..2806
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2826..2835
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2835..2858
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2865..2885
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2910..7010
                     /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             2910..7010
                     /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             7011..7058
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7074..7130
                     /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             7131..7730
                     /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(7131..7150)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7627..7647
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7746..8334
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7799..7819)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8217..8228)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8406..8639
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8717..8838
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8754..8773)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8808..8827
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8878..9013
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9033..9052)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9034..9052)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9062..9079)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9062..9078)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9071..9093)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9220..9675
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9307..9326)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9517..9538
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9701..9805
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9806..9874,9875..10666)
                     /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(10024..10043)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10837..11425
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11326..11345
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(11579..11590,1..6)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
       61 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
      121 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
      181 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
      241 gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga gctgcaagct
      301 taatgtagtc ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg agttagcaac
      361 atgccttaca aggagagaaa aagcaccgtg catgccgatt ggtggaagta aggtggtacg
      421 atcgtgcctt attaggaagg caacagacgg gtctgacatg gattggacga accactgaat
      481 tgccgcattg cagagatatt gtatttaagt gcctagctcg atacataaac gggtctctct
      541 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
      601 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
      661 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
      721 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
      781 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      841 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      901 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      961 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
     1021 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
     1081 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
     1141 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
     1201 taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg
     1261 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1321 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1381 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     1441 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1501 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1561 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1621 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1681 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     1741 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     1801 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     1861 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     1921 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     1981 ttcagaccca cctcccaacc ccgaggggac ccagagaggg cctatttccc atgattcctt
     2041 catatttgca tatacgatac aaggctgtta gagagataat tagaattaat ttgactgtaa
     2101 acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg
     2161 cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt
     2221 tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgtcctt tatcacatac
     2281 atgtggtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
     2341 aaagtggcac cgagtcggtg cttttttgaa ttctagatct tgagacaaat ggcagtattc
     2401 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     2461 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     2521 aattttcggg tttattacag ggacagcaga gatccacttt ggcgccggct cgagtggctc
     2581 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     2641 ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     2701 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     2761 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtgt cgtgacgcgg
     2821 gatccgccac catggattac aaagacgatg acgataagat ggccccaaag aagaagcgga
     2881 aggtcggtat ccacggagtc ccagcagccg acaagaagta cagcatcggc ctggacatcg
     2941 gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat
     3001 tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc
     3061 tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat
     3121 acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca
     3181 aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga
     3241 agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt
     3301 accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc
     3361 ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg
     3421 gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct
     3481 acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc
     3541 tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg
     3601 agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg acccccaact
     3661 tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg
     3721 acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg
     3781 ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga
     3841 tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc
     3901 tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct
     3961 tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt
     4021 tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga
     4081 agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc
     4141 accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat
     4201 tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg
     4261 tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa
     4321 ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca
     4381 tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca
     4441 gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg
     4501 agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc
     4561 tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa
     4621 tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg
     4681 gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa
     4741 acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga
     4801 tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga
     4861 agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg
     4921 acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa
     4981 acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc
     5041 aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg
     5101 ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg
     5161 gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga
     5221 agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg
     5281 gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt
     5341 acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc
     5401 ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag gacgactcca
     5461 tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct
     5521 ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga
     5581 ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg
     5641 ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg
     5701 cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg
     5761 aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt
     5821 tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg
     5881 tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg
     5941 actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg
     6001 ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc
     6061 tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga
     6121 tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag
     6181 tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc
     6241 ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg
     6301 gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg
     6361 gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa
     6421 gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa
     6481 aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga
     6541 gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat
     6601 atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata
     6661 atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc
     6721 agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt
     6781 ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc
     6841 tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg
     6901 accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca
     6961 tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac aagcgtcctg
     7021 ctgctactaa gaaagctggt caagctaaga aaaagaaagc tagcggcagc ggcgccacca
     7081 acttcagcct gctgaagcag gccggcgacg tggaggagaa ccccggcccc atgaccgagt
     7141 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     7201 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     7261 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     7321 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     7381 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     7441 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     7501 tcctggccac cgtcggagtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     7561 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     7621 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     7681 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga acgcgttaag
     7741 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     7801 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     7861 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     7921 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     7981 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     8041 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     8101 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     8161 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     8221 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     8281 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcgtcgac
     8341 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
     8401 gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg
     8461 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     8521 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     8581 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     8641 tacgtatagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     8701 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     8761 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     8821 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc
     8881 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     8941 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     9001 ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
     9061 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     9121 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     9181 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     9241 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     9301 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     9361 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     9421 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     9481 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     9541 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     9601 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     9661 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     9721 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     9781 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     9841 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     9901 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     9961 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
    10021 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
    10081 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
    10141 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
    10201 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
    10261 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
    10321 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
    10381 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
    10441 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
    10501 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
    10561 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
    10621 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
    10681 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
    10741 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
    10801 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
    10861 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
    10921 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
    10981 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
    11041 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
    11101 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
    11161 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
    11221 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
    11281 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
    11341 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
    11401 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
    11461 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
    11521 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
    11581 cgagtcagtg
//
