LOCUS       sequence-190168-       13013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressed Cas9 with sgRNA targeting Beclin1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13013)
  AUTHORS   Imam S, Talley S, Nelson RS, Dharan A, O'Connor C, Hope TJ, Campbell
            EM
  TITLE     TRIM5alpha degradation via autophagy is not required for retroviral 
            restriction.
  JOURNAL   J Virol. 2016 Jan 13. pii: JVI.03033-15.
  PUBMED    26764007
REFERENCE   2  (bases 1 to 13013)
  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: "J Virol. 
            2016 Jan 13. pii: JVI.03033-15."
COMMENT     Sequence Label: LentiCRISPRv2-Beclin1
FEATURES             Location/Qualifiers
     source          1..13013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(6..25)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     252..272
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(281..709)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(391..412)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     603..622
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    755..979
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     968..985
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             1011..1244
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    1316..1904
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1422..1433
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1831..1851
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1920..2516)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2003..2023)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             complement(2517..2573)
                     /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             complement(2583..2606)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(2607..2654)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(2655..6758)
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(2655..6758)
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     regulatory      6755..6764
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6783..6994)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_RNA        complement(7056..7131)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(7153..7401)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(7211..7230)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(7381..7401)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    7452..7569
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(7867..7911)
                     /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"
     misc_feature    8096..8329
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    8822..8947
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             8994..9174
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(9192..9390)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(9217..9237)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        9392..9771
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     9943..9962
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        10037..10141
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10142..10210,10211..11002)
                     /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(10360..10379)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11173..11761
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11662..11681
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11915..11932
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12049..12070
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12085..12090,12091..12108,12109..12115)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12123..12139
                     /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     12128..12150
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12147..12163
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12147..12163
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    12212..12333
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12223..12242)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     12277..12296
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(12463..12837)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(12856..12903)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(12951..13013,1..267))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(12965..13013,1..87)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
       61 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
      121 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
      181 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
      241 ggactttcca caccctaact gacacacatt ccacagaatt aattcgcgtt aaatttttgt
      301 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
      361 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
      421 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
      481 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
      541 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
      601 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
      661 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtg gggatacccc
      721 ctagagcccc agctggttct ttccgcctca gaagccatag agcccaccgc atccccagca
      781 tgcctgctat tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga
      841 atagaatgac acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca
      901 gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc
      961 tggcaactag aaggcacagt cgaggctgat cagcgggttt aaacgggccc tgctagagat
     1021 tttccacact gactaaaagg gtctgaggga tctctagtta ccagagtcac acaacagacg
     1081 ggcacacact acttgaagca ctcaaggcaa gctttattga ggcttaagca gtgggttccc
     1141 tagttagcca gagagctccc aggctcagat ctggtctaac cagagagacc cagtacaagc
     1201 aaaaagcaga tcttgtcttc gttgggagtg aattagccct tccagtcccc ccttttcttt
     1261 taaaaagtgg ctaagatcta cagctgcctt gtaagtcatt ggtcttaaag tcgacgcggg
     1321 gaggcggccc aaagggagat ccgactcgtc tgagggcgaa ggcgaagacg cggaagaggc
     1381 cgcagagccg gcagcaggcc gcgggaagga aggtccgctg gattgagggc cgaagggacg
     1441 tagcagaagg acgtcccgcg cagaatccag gtggcaacac aggcgagcag ccaaggaaag
     1501 gacgatgatt tccccgacaa caccacggaa ttgtcagtgc ccaacagccg agcccctgtc
     1561 cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg caatagggag ggggaaagcg
     1621 aaagtcccgg aaaggagctg acaggtggtg gcaatgcccc aaccagtggg ggttgcgtca
     1681 gcaaacacag tgcacaccac gccacgttgc ctgacaacgg gccacaactc ctcataaaga
     1741 gacagcaacc aggatttata caaggaggag aaaatgaaag ccatacggga agcaatagca
     1801 tgatacaaag gcattaaagc agcgtatcca catagcgtaa aaggagcaac atagttaaga
     1861 ataccagtca atctttcaca aattttgtaa tccagaggtt gattgtcgac ttaacgcgtt
     1921 caggcaccgg gcttgcgggt catgcaccag gtgcgcggtc cttcgggcac ctcgacgtcg
     1981 gcggtgacgg tgaagccgag ccgctcgtag aaggggaggt tgcggggcgc ggaggtctcc
     2041 aggaaggcgg gcaccccggc gcgctcggcc gcctccactc cggggagcac gacggcgctg
     2101 cccagaccct tgccctggtg gtcgggcgag actccgacgg tggccaggaa ccacgcgggc
     2161 tccttgggcc ggtgcggcgc caggaggcct tccatctgtt gctgcgcggc cagccgggaa
     2221 ccgctcaact cggccatgcg cgggccgatc tcggcgaaca ccgcccccgc ttcgacgctc
     2281 tccggcgtgg tccagaccgc caccgcggcg ccgtcgtccg cgacccacac cttgccgatg
     2341 tcgagcccga cgcgcgtgag gaagagttct tgcagctcgg tgacccgctc gatgtggcgg
     2401 tccggatcga cggtgtggcg cgtggcgggg tagtcggcga acgcggcggc gagggtgcgt
     2461 acggccctgg ggacgtcgtc gcgggtggcg aggcgcaccg tgggcttgta ctcggtcggt
     2521 ccaggattct cttcgacatc tccggcttgt ttcagcagag agaagtttgt tgcgccggat
     2581 cccttatcgt catcgtcttt gtaatctttc ttcttcttag cctgtccagc cttctttgtg
     2641 gcggcaggtc gcttgtcgcc tcccagctga gacaggtcga tccgtgtctc gtacaggccg
     2701 gtgatgctct ggtggatcag ggtggcgtcc agcacctctt tggtgctggt gtacctcttc
     2761 cggtcgatgg tggtgtcaaa gtacttgaag gcggcagggg ctcccagatt ggtcagggta
     2821 aacaggtgga tgatattctc ggcctgctct ctgatgggct tatcccggtg cttgttgtag
     2881 gcggacagca ctttgtccag attagcgtcg gccaggatca ctctcttgga gaactcgctg
     2941 atctgctcga tgatctcgtc caggtagtgc ttgtgctgtt ccacaaacag ctgtttctgc
     3001 tcattatcct cgggggagcc cttcagcttc tcatagtggc tggccaggta caggaagttc
     3061 acatatttgg agggcagggc cagttcgttt cccttctgca gttcgccggc agaggccagc
     3121 attctcttcc ggccgttttc cagctcgaac agggagtact taggcagctt gatgatcagg
     3181 tcctttttca cttctttgta gcccttggct tccagaaagt cgatgggatt cttctcgaag
     3241 ctgcttcttt ccatgatggt gatccccagc agctctttca cactcttcag tttcttggac
     3301 ttgccctttt ccactttggc caccaccagc acagaatagg ccacggtggg gctgtcgaag
     3361 ccgccgtact tcttagggtc ccagtccttc tttctggcga tcagcttatc gctgttcctc
     3421 ttgggcagga tagactcttt gctgaagccg cctgtctgca cctcggtctt tttcacgata
     3481 ttcacttggg gcatgctcag cactttccgc acggtggcaa aatcccggcc cttatcccac
     3541 acgatctccc cggtttcgcc gtttgtctcg atcagaggcc gcttccggat ctcgccgttg
     3601 gccagggtaa tctcggtctt gaaaaagttc atgatgttgc tgtagaagaa gtacttggcg
     3661 gtagccttgc cgatttcctg ctcgctcttg gcgatcatct tccgcacgtc gtacaccttg
     3721 tagtcgccgt acacgaactc gctttccagc ttagggtact ttttgatcag ggcggttccc
     3781 acgacggcgt tcaggtaggc gtcgtgggcg tggtggtagt tgttgatctc gcgcactttg
     3841 taaaactgga aatccttccg gaaatcggac accagcttgg acttcagggt gatcactttc
     3901 acttcccgga tcagcttgtc attctcgtcg tacttagtgt tcatccggga gtccaggatc
     3961 tgtgccacgt gctttgtgat ctgccgggtt tccaccagct gtctcttgat gaagccggcc
     4021 ttatccagtt cgctcaggcc gcctctctcg gccttggtca gattgtcgaa ctttctctgg
     4081 gtaatcagct tggcgttcag cagctgccgc cagtagttct tcatcttctt cacgacctct
     4141 tcggagggca cgttgtcgct cttgccccgg ttcttgtcgc ttctggtcag caccttgttg
     4201 tcgatggagt cgtccttcag aaagctctga ggcacgatat ggtccacatc gtagtcggac
     4261 agccggttga tgtccagttc ctggtccacg tacatatccc gcccattctg caggtagtac
     4321 aggtacagct tctcgttctg cagctgggtg ttttccacgg ggtgttcttt caggatctgg
     4381 ctgcccagct ctttgatgcc ctcttcgatc cgcttcattc tctcgcggct gttcttctgt
     4441 cccttctggg tggtctggtt ctctctggcc atttcgatca cgatgttctc gggcttgtgc
     4501 cggcccatca ctttcacgag ctcgtccacc accttcactg tctgcaggat gcccttctta
     4561 atggcggggc tgccggccag attggcaatg tgctcgtgca ggctatcgcc ctggccggac
     4621 acctgggctt tctggatgtc ctctttaaag gtcaggctgt cgtcgtggat cagctgcatg
     4681 aagtttctgt tggcgaagcc gtcggacttc aggaaatcca ggattgtctt gccggactgc
     4741 ttgtcccgga tgccgttgat cagcttccgg ctcagcctgc cccagccggt gtatctccgc
     4801 cgcttcagct gcttcatcac tttgtcgtcg aacaggtggg cataggtttt cagccgttcc
     4861 tcgatcatct ctctgtcctc aaacagtgtc agggtcagca cgatatcttc cagaatgtcc
     4921 tcgttttcct cattgtccag gaagtccttg tccttgataa ttttcagcag atcgtggtat
     4981 gtgcccaggg aggcgttgaa ccgatcttcc acgccggaga tttccacgga gtcgaagcac
     5041 tcgattttct tgaagtagtc ctctttcagc tgcttcacgg tcactttccg gttggtcttg
     5101 aacagcaggt ccacgatggc ctttttctgc tcgccgctca ggaaggcggg ctttctcatt
     5161 ccctcggtca cgtatttcac tttggtcagc tcgttataca cggtgaagta ctcgtacagc
     5221 aggctgtgct tgggcagcac cttctcgttg ggcaggttct tatcgaagtt ggtcatccgc
     5281 tcgatgaagc tctgggcgga agcgcccttg tccaccactt cctcgaagtt ccagggggtg
     5341 atggtttcct cgctctttct ggtcatccag gcgaatctgc tgtttcccct ggccagaggg
     5401 cccacgtagt aggggatgcg gaaggtcagg atcttctcga tcttttcccg gttgtccttc
     5461 aggaatgggt aaaaatcttc ctgccgccgc agaatggcgt gcagctctcc caggtggatc
     5521 tggtggggga tgctgccgtt gtcgaaggtc cgctgcttcc gcagcaggtc ctctctgttc
     5581 agcttcacga gcagttcctc ggtgccgtcc atcttttcca ggatgggctt gatgaacttg
     5641 tagaactctt cctggctggc tccgccgtca atgtagccgg cgtagccgtt cttgctctgg
     5701 tcgaagaaaa tctctttgta cttctcaggc agctgctgcc gcacgagagc tttcagcagg
     5761 gtcaggtcct ggtggtgctc gtcgtatctc ttgatcatag aggcgctcag gggggccttg
     5821 gtgatctcgg tgttcactct caggatgtcg ctcagcagga tggcgtcgga caggttcttg
     5881 gcggccagaa acaggtcggc gtactggtcg ccgatctggg ccagcaggtt gtccaggtcg
     5941 tcgtcgtagg tgtccttgct cagctgcagt ttggcatcct cggccaggtc gaagttgctc
     6001 ttgaagttgg gggtcaggcc caggctcagg gcaatcaggt ttccgaacag gccattcttc
     6061 ttctcgccgg gcagctgggc gatcagattt tccagccgtc tgctcttgct cagtctggca
     6121 gacaggatgg ccttggcgtc cacgccgctg gcgttgatgg ggttttcctc gaacagctgg
     6181 ttgtaggtct gcaccagctg gatgaacagc ttgtccacgt cgctgttgtc ggggttcagg
     6241 tcgccctcga tcaggaagtg gccccggaac ttgatcatgt gggccagggc cagatagatc
     6301 agccgcaggt cggccttgtc ggtgctgtcc accagtttct ttctcaggtg gtagatggtg
     6361 gggtacttct cgtggtaggc cacctcgtcc acgatgttgc cgaagatggg gtgccgctcg
     6421 tgcttcttat cctcttccac caggaaggac tcttccagtc tgtggaagaa gctgtcgtcc
     6481 accttggcca tctcgttgct gaagatctct tgcagatagc agatccggtt cttccgtctg
     6541 gtgtatcttc ttctggcggt tctcttcagc cgggtggcct cggctgtttc gccgctgtcg
     6601 aacagcaggg ctccgatcag gttcttcttg atgctgtgcc ggtcggtgtt gcccagcacc
     6661 ttgaatttct tgctgggcac cttgtactcg tcggtgatca cggcccagcc cacagagttg
     6721 gtgccgatgt ccaggccgat gctgtacttc ttgtccatgg tggcagcgct ctagaaccgg
     6781 tcctgtgttc tggcggcaaa cccgttgcga aaaagaacgt tcacggcgac tactgcactt
     6841 atatacggtt ctcccccacc ctcgggaaaa aggcggagcc agtacacgac atcactttcc
     6901 cagtttaccc cgcgccacct tctctaggca ccggatcaat tgccgacccc tccccccaac
     6961 ttctcgggga ctgtgggcga tgtgcgctct gcccactgac gggcaccgga gccaattccc
     7021 actcctttca agacctagct agcgaattca aaaaagcacc gactcggtgc cactttttca
     7081 agttgataac ggactagcct tattttaact tgctatttct agctctaaaa cggatggtgt
     7141 ctctcgcaga tcggtgtttc gtcctttcca caagatatat aaagccaaga aatcgaaata
     7201 ctttcaagtt acggtaagca tatgatagtc cattttaaaa cataatttta aaactgcaaa
     7261 ctacccaaga aattattact ttctacgtca cgtattttgt actaatatct ttgtgtttac
     7321 agtcaaatta attctaatta tctctctaac agccttgtat cgtatatgca aatatgaagg
     7381 aatcatggga aataggccct cggtacctta attaaccaaa ctggatctct gctgtccctg
     7441 taataaaccc gaaaattttg aatttttgta atttgttttt gtaattcttt agtttgtatg
     7501 tctgttgcta ttatgtctac tattctttcc cctgcactgt accccccaat cccccctttt
     7561 cttttaaaat tgtggatgaa tactgccatt tgtctgcaga attggcgcac gcagtgccga
     7621 tccgttcact aatcgaatgg atctgtctct gtctctctct ccaccttctt cttctattcc
     7681 ttcgggcctg tcgggtcccc tcggggttgg gaggtgggtc tgaaacgata atggtgaata
     7741 tccctgccta actctattca ctatagaaag tacagcaaaa actattctta aacctaccaa
     7801 gcctcctact atcattatga ataattttat ataccacagc caatttgtta tgttaaacca
     7861 attccacaaa cttgcccatt tatctaattc caataattct tgttcattct tttcttgctg
     7921 gttttgcgat tcttcaatta aggagtgtat taagcttgtg taattgttaa tttctctgtc
     7981 ccactccatc caggtcgtgt gattccaaat ctgttccaga gatttattac tccaactagc
     8041 attccaaggc acagcagtgg tgcaaatgag ttttccagag caaccccaaa tccccaggag
     8101 ctgttgatcc tttaggtatc tttccacagc caggattctt gcctggagct gcttgatgcc
     8161 ccagactgtg agttgcaaca gatgctgttg cgcctcaata gccctcagca aattgttctg
     8221 ctgctgcact ataccagaca ataattgtct ggcctgtacc gtcagcgtca ttgacgctgc
     8281 gcccatagtg cttcctgctg ctcccaagaa cccaaggaac aaagctccta ttcccactgc
     8341 tcttttttct ctctgcacca ctcttctctt tgccttggtg ggtgctactc ctaatggttc
     8401 aatttttact actttatatt tatataattc acttctccaa ttgtccctca tatctcctcc
     8461 tccaggtctg aagatcagcg gccgcttgct gtgcggtggt cttacttttg ttttgctctt
     8521 cctctatctt gtctaaagct tccttggtgt cttttatctc tatcctttga tgcacacaat
     8581 agagggttgc tactgtatta tataatgatc taagttcttc tgatcctgtc tgaagggatg
     8641 gttgtagctg tcccagtatt tgtctacagc cttctgatgt ttctaacagg ccaggattaa
     8701 ctgcgaatcg ttctagctcc ctgcttgccc atactatatg ttttaattta tattttttct
     8761 ttccccctgg ccttaaccga attttttccc atcgcgatct aattctcccc cgcttaatac
     8821 tgacgctctc gcacccatct ctctccttct agcctccgct agtcaaaatt tttggcgtac
     8881 tcaccagtcg ccgcccctcg cctcttgccg tgcgcgcttc agcaagccga gtcctgcgtc
     8941 gagagagctc ctctggtttc cctttcgctt tcaagtccct gttcgggcgc cactgctaga
     9001 gattttccac actgactaaa agggtctgag ggatctctag ttaccagagt cacacaacag
     9061 acgggcacac actacttgaa gcactcaagg caagctttat tgaggcttaa gcagtgggtt
     9121 ccctagttag ccagagagct cccaggctca gatctggtct aaccagagag acccagtaca
     9181 ggcaaaacgc gctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca
     9241 atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac
     9301 tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac
     9361 gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga
     9421 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
     9481 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
     9541 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
     9601 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc
     9661 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa
     9721 ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt caacgcgtat
     9781 atctggcccg tacatcgcga agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc
     9841 aattgtcggt caagccttgc cttgttgtag cttaaatttt gctcgcgcac tactcagcga
     9901 cctccaacac acaagcaggg agcagatact ggcttaacta tgcggcatca gagcagattg
     9961 tactgagagt gcaccatagg ggatcgggag atctcccgat ccgtcgacgt caggtggcac
    10021 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
    10081 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
    10141 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
    10201 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
    10261 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
    10321 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
    10381 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
    10441 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
    10501 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
    10561 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
    10621 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
    10681 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
    10741 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
    10801 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
    10861 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
    10921 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
    10981 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
    11041 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
    11101 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
    11161 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
    11221 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
    11281 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
    11341 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
    11401 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
    11461 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
    11521 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
    11581 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
    11641 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
    11701 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
    11761 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
    11821 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
    11881 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
    11941 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
    12001 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
    12061 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
    12121 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
    12181 ctagctagag gtcgacggta tacagacatg ataagataca ttgatgagtt tggacaaacc
    12241 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
    12301 tttgtaacca ttataagctg caataaacaa gttggggtgg gcgaagaact ccagcatgag
    12361 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
    12421 ttcatagaag gcggcggtgg aatcgaaatc tcgtagcacg tgtcagtcct gctcctcggc
    12481 cacgaagtgc acgcagttgc cggccgggtc gcgcagggcg aactcccgcc cccacggctg
    12541 ctcgccgatc tcggtcatgg ccggcccgga ggcgtcccgg aagttcgtgg acacgacctc
    12601 cgaccactcg gcgtacagct cgtccaggcc gcgcacccac acccaggcca gggtgttgtc
    12661 cggcaccacc tggtcctgga ccgcgctgat gaacagggtc acgtcgtccc ggaccacacc
    12721 ggcgaagtcg tcctccacga agtcccggga gaacccgagc cggtcggtcc agaactcgac
    12781 cgctccggcg acgtcgcgcg cggtgagcac cggaacggca ctggtcaact tggccatggt
    12841 ttagttcctc accttgtcgt attatactat gccgatatac tatgccgatg attaattgtc
    12901 aacacgtgct gatcagatcc gaaaatggat atacaagctc ccgggagctt tttgcaaaag
    12961 cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc aga
//
