LOCUS       sequence-180843-       13012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgNf1.4 for Nf1 deletion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13012)
  AUTHORS   Song CQ, Li Y, Mou H, Moore J, Park A, Pomyen Y, Hough S, Kennedy Z,
            Fischer A, Yin H, Anderson DG, Conte D Jr, Zender L, Wang XW, 
            Thorgeirsson S, Weng Z, Xue W
  TITLE     Genome-Wide CRISPR Screen Identifies Regulators of Mitogen-Activated
            Protein Kinase as Suppressors of Liver Tumors in Mice.
  JOURNAL   Gastroenterology. 2017 Apr;152(5):1161-1173.e1. doi: 
            10.1053/j.gastro.2016.12.002. Epub 2016 Dec 10.
  PUBMED    27956228
REFERENCE   2  (bases 1 to 13012)
  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: 
            "10.1053/j.gastro.2016.12.002"; journalName: "Gastroenterology"; 
            date: "2017-04"; volume: "152"; issue: "5"; pages: "1161-1173.e1"
COMMENT     Sequence Label: lentiCRISPRv2.sgNf1.4
FEATURES             Location/Qualifiers
     source          1..13012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        270..345
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        407..618
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      637..646
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             643..4746
                     /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             643..4746
                     /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             4747..4794
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4795..4818
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4828..4884
                     /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             4885..5481
                     /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     5378..5398
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    5497..6085
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5550..5570)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5968..5979)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6157..6390
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6416..6433)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6422..6646
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6692..7120
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6779..6798)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6989..7010
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7129..7149)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7134..7463
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7314..7449
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7376..7395
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7511..7558
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7577..7951
                     /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"
     polyA_signal    8081..8202
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8118..8137)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8172..8191
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8251..8267)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8251..8267)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8264..8286)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8275..8291
                     /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)."
     promoter        complement(join(8299..8305,8306..8323,8324..8329))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8344..8365
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8482..8499)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8653..9241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8733..8752)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9412..10203,10204..10272))
                     /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     10035..10054
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10273..10377)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10452..10471)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        10643..11022
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11024..11222
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     11177..11197
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             11240..11420
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11467..11592
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12085..12318
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12503..12547
                     /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    12845..12962
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg attatccgaa ttcttagcag ttttagagct agaaatagca agttaaaata
      301 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgaattcgct
      361 agctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca
      421 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgatccgg tgcctagaga
      481 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
      541 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
      601 tttgccgcca gaacacagga ccggttctag agcgctgcca ccatggacaa gaagtacagc
      661 atcggcctgg acatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag
      721 gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac
      781 ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga
      841 accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc
      901 agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg
      961 gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg
     1021 gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc
     1081 gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc
     1141 cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc
     1201 cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg
     1261 gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc
     1321 gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc cctgagcctg
     1381 ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg
     1441 agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac
     1501 gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg
     1561 agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac
     1621 gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag
     1681 tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga
     1741 gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc
     1801 gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac
     1861 aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag
     1921 gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc
     1981 cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc
     2041 agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct
     2101 tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag
     2161 gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa
     2221 gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag
     2281 gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag
     2341 gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg
     2401 ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc
     2461 ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt
     2521 gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa
     2581 gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg
     2641 atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac
     2701 ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag
     2761 gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat
     2821 ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag
     2881 ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag
     2941 aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag
     3001 ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg
     3061 cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag
     3121 gaactggaca tcaaccggct gtccgactac gatgtggacc atatcgtgcc tcagagcttt
     3181 ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag
     3241 agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg
     3301 ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc
     3361 ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag
     3421 atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat
     3481 gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc
     3541 cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac
     3601 gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc
     3661 gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag
     3721 caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc
     3781 aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac
     3841 ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg
     3901 ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc
     3961 aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg
     4021 gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg
     4081 gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc
     4141 accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc
     4201 tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg
     4261 gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg
     4321 gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag
     4381 ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg
     4441 gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat
     4501 ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc
     4561 gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac
     4621 tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc
     4681 ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga
     4741 ggcgacaagc gacctgccgc cacaaagaag gctggacagg ctaagaagaa gaaagattac
     4801 aaagacgatg acgataaggg atccggcgca acaaacttct ctctgctgaa acaagccgga
     4861 gatgtcgaag agaatcctgg accgaccgag tacaagccca cggtgcgcct cgccacccgc
     4921 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     4981 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     5041 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     5101 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     5161 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     5221 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggagt ctcgcccgac
     5281 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     5341 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     5401 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     5461 acccgcaagc ccggtgcctg aacgcgttaa gtcgacaatc aacctctgga ttacaaaatt
     5521 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     5581 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     5641 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     5701 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     5761 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     5821 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     5881 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     5941 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     6001 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     6061 atctcccttt gggccgcctc cccgcgtcga ctttaagacc aatgacttac aaggcagctg
     6121 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     6181 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     6241 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6301 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     6361 ccttttagtc agtgtggaaa atctctagca gggcccgttt aaacccgctg atcagcctcg
     6421 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     6481 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     6541 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     6601 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     6661 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     6721 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     6781 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     6841 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     6901 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     6961 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     7021 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     7081 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     7141 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     7201 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     7261 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     7321 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     7381 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     7441 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     7501 atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     7561 ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga
     7621 cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga
     7681 ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga
     7741 ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta
     7801 cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac
     7861 cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg
     7921 cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc
     7981 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     8041 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     8101 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     8161 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     8221 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     8281 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     8341 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     8401 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     8461 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     8521 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     8581 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     8641 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     8701 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     8761 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     8821 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     8881 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     8941 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     9001 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     9061 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     9121 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     9181 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     9241 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     9301 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     9361 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     9421 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     9481 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     9541 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     9601 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     9661 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     9721 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     9781 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     9841 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     9901 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     9961 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
    10021 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
    10081 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
    10141 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
    10201 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
    10261 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
    10321 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
    10381 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
    10441 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
    10501 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
    10561 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
    10621 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
    10681 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
    10741 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
    10801 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
    10861 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
    10921 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
    10981 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
    11041 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
    11101 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
    11161 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
    11221 agcgcgtttt gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
    11281 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
    11341 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
    11401 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
    11461 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
    11521 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
    11581 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
    11641 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
    11701 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
    11761 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
    11821 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
    11881 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccgctgat
    11941 cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat ataaatataa
    12001 agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa gagtggtgca
    12061 gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg gagcagcagg
    12121 aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat tattgtctgg
    12181 tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc atctgttgca
    12241 actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg aaagatacct
    12301 aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt gcaccactgc
    12361 tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga atcacacgac
    12421 ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact ccttaattga
    12481 agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag ataaatgggc
    12541 aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat tattcataat
    12601 gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta tagtgaatag
    12661 agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc cgaggggacc
    12721 cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca gatccattcg
    12781 attagtgaac ggatcggcac tgcgtgcgcc aattctgcag acaaatggca gtattcatcc
    12841 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
    12901 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
    12961 ttcgggttta ttacagggac agcagagatc cagtttggtt aattaaggta cc
//
