LOCUS       sequence-327642-       11589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For CRISPR knockout of miR-144~451 linker region by lentiviral 
            delivery of Cas9 and linker gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11589)
  AUTHORS   Fang W, Bartel DP
  TITLE     MicroRNA Clustering Assists Processing of Suboptimal MicroRNA 
            Hairpins through the Action of the ERH Protein.
  JOURNAL   Mol Cell. 2020 Apr 16;78(2):289-302.e6. doi: 
            10.1016/j.molcel.2020.01.026.
  PUBMED    32302541
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"; doi: 
            "10.1016/j.molcel.2020.01"; journalName: "Mol Cell"; date: 
            "2020-04-16- 16"; volume: "78"; issue: "2"; pages: "289-302.e6"
COMMENT     Sequence Label: pLentiCRISPRv1.linker_KO
FEATURES             Location/Qualifiers
     source          1..11589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     147..166
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     400..417
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    534..555
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(570..575,576..593,594..600)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    608..624
                     /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     613..635
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     632..648
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     632..648
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     667..687
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        669..687
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        715..941
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             942..1122
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1169..1294
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1787..2020
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2205..2249
                     /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        2427..2675
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2427..2447
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2598..2617
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2696..2771
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2819..2936
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3005..3216
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3236..3245
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3245..3268
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3275..3295
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3320..7420
                     /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             3320..7420
                     /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             7421..7468
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7484..7540
                     /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             7541..8140
                     /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(7541..7560)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8037..8057
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8156..8744
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8209..8229)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8627..8638)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8816..9049
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9127..9248
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9164..9183)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9218..9237
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9288..9423
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9443..9462)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9444..9462)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9472..9489)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9472..9488)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9481..9503)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9630..10085
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9717..9736)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9927..9948
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10111..10215
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10216..10284,10285..11076)
                     /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(10434..10453)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(11247..11589,1..246)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
       61 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
      121 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
      181 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
      241 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
      301 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
      361 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
      421 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
      481 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
      541 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
      601 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
      661 aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt
      721 cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa catgccttac
      781 aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac gatcgtgcct
      841 tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa ttgccgcatt
      901 gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc tggttagacc
      961 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
     1021 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     1081 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
     1141 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
     1201 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
     1261 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
     1321 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
     1381 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
     1441 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     1501 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     1561 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
     1621 cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     1681 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     1741 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     1801 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     1861 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     1921 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     1981 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     2041 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     2101 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     2161 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     2221 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     2281 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     2341 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     2401 acctcccaac cccgagggga cccagagagg gcctatttcc catgattcct tcatatttgc
     2461 atatacgata caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga
     2521 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
     2581 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct
     2641 tggctttata tatcttgtgg aaaggacgaa acaccggctg agcagagagc ttcttgtttt
     2701 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac
     2761 cgagtcggtg cttttttgaa ttctagatct tgagacaaat ggcagtattc atccacaatt
     2821 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag
     2881 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg
     2941 tttattacag ggacagcaga gatccacttt ggcgccggct cgagtggctc cggtgcccgt
     3001 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     3061 tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     3121 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     3181 gttctttttc gcaacgggtt tgccgccaga acacaggtgt cgtgacgcgg gatccgccac
     3241 catggattac aaagacgatg acgataagat ggccccaaag aagaagcgga aggtcggtat
     3301 ccacggagtc ccagcagccg acaagaagta cagcatcggc ctggacatcg gcaccaactc
     3361 tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct
     3421 gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag
     3481 cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg
     3541 gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga
     3601 cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg
     3661 gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat
     3721 ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta
     3781 tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa
     3841 ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct
     3901 gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag
     3961 actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa
     4021 tggcctgttc ggaaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa
     4081 cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct
     4141 ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa
     4201 cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc
     4261 ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct
     4321 gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag
     4381 caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt
     4441 catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag
     4501 agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca
     4561 cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga
     4621 caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct
     4681 ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc
     4741 ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat
     4801 gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta
     4861 cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag
     4921 aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac
     4981 caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt
     5041 cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca
     5101 cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat
     5161 tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg
     5221 gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag
     5281 atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc
     5341 cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca
     5401 gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg
     5461 ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa
     5521 gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa
     5581 gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa
     5641 gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat
     5701 cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta
     5761 cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga
     5821 ctacgatgtg gaccatatcg tgcctcagag ctttctgaag gacgactcca tcgacaacaa
     5881 ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct ccgaagaggt
     5941 cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga ttacccagag
     6001 aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg
     6061 cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct
     6121 ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg aagtgaaagt
     6181 gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt
     6241 gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac
     6301 cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt
     6361 gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa
     6421 gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg
     6481 cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga tcgtgtggga
     6541 taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt
     6601 gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa
     6661 cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga
     6721 cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa
     6781 gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga
     6841 gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat
     6901 catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc
     6961 ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt
     7021 cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa
     7081 acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc agatcagcga
     7141 gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa
     7201 caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct
     7261 gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag
     7321 gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct
     7381 gtacgagaca cggatcgacc tgtctcagct gggaggcgac aagcgtcctg ctgctactaa
     7441 gaaagctggt caagctaaga aaaagaaagc tagcggcagc ggcgccacca acttcagcct
     7501 gctgaagcag gccggcgacg tggaggagaa ccccggcccc atgaccgagt acaagcccac
     7561 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     7621 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     7681 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     7741 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     7801 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     7861 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     7921 cgtcggagtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     7981 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     8041 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     8101 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga acgcgttaag tcgacaatca
     8161 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     8221 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     8281 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     8341 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     8401 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     8461 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     8521 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     8581 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     8641 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     8701 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcgtcgac tttaagacca
     8761 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     8821 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg
     8881 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     8941 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     9001 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tacgtatagt
     9061 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     9121 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     9181 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     9241 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc ccgcccctaa
     9301 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     9361 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     9421 gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt
     9481 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     9541 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     9601 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     9661 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     9721 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     9781 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     9841 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     9901 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     9961 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
    10021 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
    10081 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
    10141 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
    10201 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
    10261 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
    10321 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
    10381 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
    10441 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
    10501 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
    10561 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
    10621 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
    10681 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
    10741 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
    10801 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
    10861 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
    10921 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
    10981 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
    11041 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
    11101 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
    11161 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
    11221 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
    11281 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
    11341 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
    11401 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
    11461 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
    11521 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
    11581 acacagccc
//
