LOCUS       sequence-328999-       11590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a sgRNA targeting stop codon of human FMR1 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11590)
  AUTHORS   Li M, Shin J, Risgaard RD, Parries MJ, Wang J, Chasman D, Liu S, Roy
            S, Bhattacharyya A, Zhao X
  TITLE     Identification of FMR1-regulated molecular networks in human 
            neurodevelopment.
  JOURNAL   Genome Res. 2020 Mar;30(3):361-374. doi: 10.1101/gr.251405.119. Epub
            2020 Mar 16.
  PUBMED    32179589
REFERENCE   2  (bases 1 to 11590)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1101/gr.251405.119"; journalName: "Genome Res"; date: "2020-03";
            volume: "30"; issue: "3"; pages: "361-374"
COMMENT     Sequence Label: 49535-sgFMR1_E17A
FEATURES             Location/Qualifiers
     source          1..11590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    95..116
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(131..136,137..154,155..161)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    169..185
                     /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     174..196
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     193..209
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     193..209
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     228..248
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        230..248
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        276..502
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             503..683
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    730..855
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1348..1581
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1766..1810
                     /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        1988..2236
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1988..2008
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2159..2178
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2258..2333
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2381..2498
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2567..2778
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2798..2807
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2807..2830
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2837..2857
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2882..6982
                     /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             2882..6982
                     /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             6983..7030
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7046..7102
                     /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             7103..7702
                     /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(7103..7122)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7599..7619
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7718..8306
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7771..7791)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8189..8200)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8378..8611
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8689..8810
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8726..8745)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8780..8799
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8850..8985
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9005..9024)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9006..9024)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9034..9051)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9034..9050)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9043..9065)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9192..9647
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9279..9298)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9489..9510
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9673..9777
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9778..9846,9847..10638)
                     /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(9996..10015)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10809..11397
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11298..11317
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11551..11568
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
       61 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      121 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      181 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
      241 actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta
      301 gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg
      361 tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac
      421 gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa
      481 gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct
      541 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
      601 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
      661 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
      721 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
      781 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
      841 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
      901 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
      961 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata
     1021 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
     1081 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
     1141 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct
     1201 gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata
     1261 taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt
     1321 gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc
     1381 aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc
     1441 tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt
     1501 gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata
     1561 cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac
     1621 tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac
     1681 gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat
     1741 tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg
     1801 ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa aattattcat
     1861 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa
     1921 tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg
     1981 acccagagag ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt
     2041 tagagagata attagaatta atttgactgt aaacacaaag atattagtac aaaatacgtg
     2101 acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga
     2161 ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg
     2221 gaaaggacga aacaccgtag ggtactccat tcacgaggtt ttagagctag aaatagcaag
     2281 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg
     2341 aattctagat cttgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
     2401 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     2461 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
     2521 gagatccact ttggcgccgg ctcgagtggc tccggtgccc gtcagtgggc agagcgcaca
     2581 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga
     2641 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     2701 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     2761 tttgccgcca gaacacaggt gtcgtgacgc gggatccgcc accatggatt acaaagacga
     2821 tgacgataag atggccccaa agaagaagcg gaaggtcggt atccacggag tcccagcagc
     2881 cgacaagaag tacagcatcg gcctggacat cggcaccaac tctgtgggct gggccgtgat
     2941 caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca
     3001 cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc
     3061 cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta
     3121 tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct tccacagact
     3181 ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa
     3241 catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa
     3301 actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat
     3361 gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt
     3421 ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat
     3481 caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg
     3541 gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct
     3601 gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga
     3661 tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca
     3721 gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct
     3781 gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat
     3841 gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca
     3901 gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg
     3961 ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga
     4021 aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa
     4081 gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc
     4141 cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga
     4201 gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag
     4261 attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt
     4321 ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa
     4381 cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta
     4441 taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag
     4501 cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt
     4561 gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc
     4621 cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat
     4681 caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct
     4741 gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca
     4801 cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag
     4861 gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga
     4921 tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag
     4981 cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca
     5041 cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt
     5101 gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat
     5161 cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat
     5221 gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt
     5281 ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga
     5341 tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggaccatat
     5401 cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga ccagaagcga
     5461 caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa
     5521 ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac
     5581 caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct
     5641 ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac
     5701 taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa
     5761 gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta
     5821 ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta
     5881 ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat
     5941 gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa
     6001 catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc
     6061 tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc gggattttgc
     6121 caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca
     6181 gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata agctgatcgc
     6241 cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta
     6301 ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa
     6361 agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt
     6421 tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta
     6481 ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca
     6541 gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca
     6601 ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca
     6661 gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat
     6721 cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc gggataagcc
     6781 catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc
     6841 tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga
     6901 ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga
     6961 cctgtctcag ctgggaggcg acaagcgtcc tgctgctact aagaaagctg gtcaagctaa
     7021 gaaaaagaaa gctagcggca gcggcgccac caacttcagc ctgctgaagc aggccggcga
     7081 cgtggaggag aaccccggcc ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     7141 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     7201 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     7261 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     7321 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     7381 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     7441 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggag tctcgcccga
     7501 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     7561 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     7621 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     7681 gacccgcaag cccggtgcct gaacgcgtta agtcgacaat caacctctgg attacaaaat
     7741 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     7801 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     7861 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     7921 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     7981 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     8041 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     8101 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     8161 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     8221 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     8281 gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct
     8341 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     8401 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     8461 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     8521 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     8581 cccttttagt cagtgtggaa aatctctagc agtacgtata gtagttcatg tcatcttatt
     8641 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
     8701 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     8761 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     8821 ctctagctat cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     8881 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     8941 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa
     9001 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga
     9061 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     9121 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     9181 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     9241 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     9301 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     9361 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     9421 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     9481 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     9541 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     9601 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     9661 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     9721 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     9781 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     9841 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     9901 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     9961 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
    10021 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
    10081 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
    10141 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
    10201 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
    10261 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
    10321 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
    10381 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
    10441 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
    10501 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
    10561 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
    10621 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
    10681 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
    10741 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
    10801 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
    10861 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
    10921 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
    10981 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
    11041 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
    11101 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
    11161 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
    11221 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
    11281 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
    11341 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
    11401 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
    11461 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
    11521 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
    11581 cgcccaatac
//
