LOCUS       sequence-321912-       11589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral vector expressing Cas9 and an sgRNA targeting MPC2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11589)
  AUTHORS   Rossiter NJ, Huggler KS, Adelmann CH, Keys HR, Soens RW, Sabatini 
            DM, Cantor JR
  TITLE     CRISPR screens in physiologic medium reveal conditionally essential 
            genes in human cells.
  JOURNAL   Cell Metab. 2021 Feb 23. pii: S1550-4131(21)00061-9. doi: 
            10.1016/j.cmet.2021.02.005.
  PUBMED    33651980
REFERENCE   2  (bases 1 to 11589)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Metab.
            2021 Feb 23. pii: S1550-4131(21)00061-9. doi: 
            10.1016/j.cmet.2021.02.005."
COMMENT     Sequence Label: pLentiCRISPR-v1-sgMPC2_9
FEATURES             Location/Qualifiers
     source          1..11589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             185..229
                     /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        407..655
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     407..427
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     578..597
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        676..751
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    799..916
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        985..1196
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      1216..1225
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1225..1248
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1255..1275
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1300..5400
                     /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             1300..5400
                     /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             5401..5448
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5464..5520
                     /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             5521..6120
                     /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(5521..5540)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6017..6037
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6136..6724
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6189..6209)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6607..6618)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6796..7029
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7107..7228
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7144..7163)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7198..7217
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7268..7403
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7423..7442)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7424..7442)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7452..7469)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7452..7468)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7461..7483)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7610..8065
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7697..7716)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7907..7928
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8091..8195
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8196..8264,8265..9056)
                     /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(8414..8433)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9227..9815
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9716..9735
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9969..9986
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10103..10124
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10139..10144,10145..10162,10163..10169)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10177..10193
                     /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     10182..10204
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10201..10217
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10201..10217
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10236..10256
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10238..10256
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        10284..10510
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             10511..10691
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    10738..10863
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11356..11589
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
       61 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
      121 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
      181 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
      241 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
      301 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
      361 accattatcg tttcagaccc acctcccaac cccgagggga cccagagagg gcctatttcc
      421 catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttagaattaa
      481 tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct
      541 tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac
      601 ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccgtgta
      661 ggggttggtg tgtgcgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
      721 tcaacttgaa aaagtggcac cgagtcggtg cttttttgaa ttctagatct tgagacaaat
      781 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
      841 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
      901 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccacttt ggcgccggct
      961 cgagtggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
     1021 tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg
     1081 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
     1141 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtgt
     1201 cgtgacgcgg gatccgccac catggattac aaagacgatg acgataagat ggccccaaag
     1261 aagaagcgga aggtcggtat ccacggagtc ccagcagccg acaagaagta cagcatcggc
     1321 ctggacatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc
     1381 agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc
     1441 ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc
     1501 agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac
     1561 gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa
     1621 gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac
     1681 cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag
     1741 gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc
     1801 ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg
     1861 gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc
     1921 aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag
     1981 ctgcccggcg agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg
     2041 acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag
     2101 gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac
     2161 ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg
     2221 aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac
     2281 caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa
     2341 gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc
     2401 caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa
     2461 ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc
     2521 agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat
     2581 ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc
     2641 ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag
     2701 agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc
     2761 cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg
     2821 cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa
     2881 tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc
     2941 gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac
     3001 ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac
     3061 gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac
     3121 aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac
     3181 agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg
     3241 aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac
     3301 ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc
     3361 gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc
     3421 cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc
     3481 ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg
     3541 aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag
     3601 accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc
     3661 aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac
     3721 gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg
     3781 gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag
     3841 gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac
     3901 aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac
     3961 gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg
     4021 agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca
     4081 aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag
     4141 ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag
     4201 gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac
     4261 ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc
     4321 gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa
     4381 atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc
     4441 gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa
     4501 accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc
     4561 atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag
     4621 tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct
     4681 aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa
     4741 gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc
     4801 atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa
     4861 gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac
     4921 ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg
     4981 ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc
     5041 cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag
     5101 atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac
     5161 aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat
     5221 atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac
     5281 accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc
     5341 caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac
     5401 aagcgtcctg ctgctactaa gaaagctggt caagctaaga aaaagaaagc tagcggcagc
     5461 ggcgccacca acttcagcct gctgaagcag gccggcgacg tggaggagaa ccccggcccc
     5521 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     5581 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     5641 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     5701 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     5761 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     5821 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     5881 cccgcgtggt tcctggccac cgtcggagtc tcgcccgacc accagggcaa gggtctgggc
     5941 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     6001 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     6061 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     6121 acgcgttaag tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     6181 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     6241 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     6301 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     6361 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     6421 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     6481 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     6541 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     6601 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     6661 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     6721 ccgcgtcgac tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     6781 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat ctgctttttg
     6841 cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     6901 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     6961 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     7021 tctctagcag tacgtatagt agttcatgtc atcttattat tcagtattta taacttgcaa
     7081 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat
     7141 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     7201 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac
     7261 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     7321 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     7381 gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat
     7441 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     7501 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     7561 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     7621 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     7681 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     7741 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     7801 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     7861 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     7921 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     7981 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     8041 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     8101 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     8161 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     8221 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     8281 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     8341 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     8401 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     8461 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     8521 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     8581 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     8641 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     8701 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     8761 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     8821 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     8881 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     8941 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     9001 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     9061 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     9121 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     9181 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     9241 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     9301 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     9361 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     9421 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     9481 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     9541 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     9601 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     9661 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     9721 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     9781 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     9841 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     9901 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     9961 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
    10021 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
    10081 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
    10141 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
    10201 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
    10261 aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt cttgcaacat
    10321 ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt
    10381 ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg
    10441 gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt gcctagctcg
    10501 atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
    10561 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
    10621 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
    10681 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc
    10741 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg
    10801 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc
    10861 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg
    10921 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc
    10981 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta
    11041 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc
    11101 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata
    11161 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc
    11221 tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa
    11281 aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa
    11341 aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat
    11401 gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca
    11461 gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt
    11521 ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca
    11581 acagctcct
//
