LOCUS       sequence-309998-       11593 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of Cas9 and SB100.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11593)
  TITLE     Lenitviral sgRNA and Cas9 constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11593)
  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: 
            "Unpublished"
COMMENT     Sequence Label: Lenti Cas9/SB100
FEATURES             Location/Qualifiers
     source          1..11593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     134..154
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        136..154
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        182..408
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             409..589
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    636..761
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1254..1487
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1672..1716
                     /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    1900..2017
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2086..2297
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     promoter        2312..2330
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2338..2347
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2344..3363
                     /codon_start=1
                     /product="hyperactive variant of the Sleeping Beauty 
                     transposase (Mates et al., 2009)"
                     /label=SB100X
                     /translation="MGKSKEISQDLRKRIVDLHKSGSSLGAISKRLAVPRSSVQTIVRK
                     YKHHGTTQPSYRSGRRRVLSPRDERTLVRKVQINPRTTAKDLVKMLEETGTKVSISTVK
                     RVLYRHNLKGHSARKKPLLQNRHKKARLRFATAHGDKDRTFWRNVLWSDETKIELFGHN
                     DHRYVWRKKGEACKPKNTIPTVKHGGGSIMLWGCFAAGGTGALHKIDGIMDAVQYVDIL
                     KQHLKTSVRKLKLGRKWVFQHDNDPKHTSKVVAKWLKDNKVKVLEWPSQSPDLNPIENL
                     WAELKKRVRARRPTNLTQLHQLCQEEWAKIHPNYCGKLVEGYPKRLTQVKQFKGNATKY
                     "
     CDS             3364..3420
                     /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             3421..7524
                     /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             3421..7524
                     /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             7525..7572
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7573..7596
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    7627..8215
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7680..7700)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8098..8109)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8287..8520
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8598..8719
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8635..8654)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8689..8708
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8759..8894
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(8915..8933)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8943..8960)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8943..8959)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8952..8974)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9101..9556
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9188..9207)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9398..9419
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9582..9686
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9687..9755,9756..10547)
                     /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(9905..9924)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10718..11306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11207..11226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11460..11477
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta
      181 atgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat
      241 gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat
      301 cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg
      361 ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg
      421 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
      481 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
      541 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
      601 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
      661 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
      721 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
      781 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
      841 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
      901 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
      961 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
     1021 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
     1081 agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac
     1141 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     1201 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     1261 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     1321 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     1381 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
     1441 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
     1501 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     1561 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     1621 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     1681 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     1741 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     1801 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     1861 cagacccacc tcccaacccc gaggggaccc agaccacaat tttaaaagaa aaggggggat
     1921 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     1981 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2041 agatccactt tggcgccggc tcgagtggct ccggtgcccg tcagtgggca gagcgcacat
     2101 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa
     2161 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     2221 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     2281 ttgccgccag aacacaggtg tcgtgacgcg gtaatacgac tcactatagg gactagtgcc
     2341 accatgggaa aatcaaaaga aatcagccaa gacctcagaa aaagaattgt agacctccac
     2401 aagtctggtt catccttggg agcaatttcc aaacgcctgg cggtaccacg ttcatctgta
     2461 caaacaatag tacgcaagta taaacaccat gggaccacgc agccgtcata ccgctcagga
     2521 aggagacgcg ttctgtctcc tagagatgaa cgtactttgg tgcgaaaagt gcaaatcaat
     2581 cccagaacaa cagcaaagga ccttgtgaag atgctggagg aaacaggtac aaaagtatct
     2641 atatccacag taaaacgagt cctatatcga cataacctga aaggccactc agcaaggaag
     2701 aagccactgc tccaaaaccg acataagaaa gccagactac ggtttgcaac tgcacatggg
     2761 gacaaagatc gtactttttg gagaaatgtc ctctggtctg atgaaacaaa aatagaactg
     2821 tttggccata atgaccatcg ttatgtttgg aggaagaagg gggaggcttg caagccgaag
     2881 aacaccatcc caaccgtgaa gcacgggggt ggcagcatca tgttgtgggg gtgctttgct
     2941 gcaggaggga ctggtgcact tcacaaaata gatggcatca tggacgcggt gcagtatgtg
     3001 gatatattga agcaacatct caagacatca gtcaggaagt taaagcttgg tcgcaaatgg
     3061 gtcttccaac acgacaatga ccccaagcat acttccaaag ttgtggcaaa atggcttaag
     3121 gacaacaaag tcaaggtatt ggagtggcca tcacaaagcc ctgacctcaa tcctatagaa
     3181 aatttgtggg cagaactgaa aaagcgtgtg cgagcaagga ggcctacaaa cctgactcag
     3241 ttacaccagc tctgtcagga ggaatgggcc aaaattcacc caaattattg tgggaagctt
     3301 gtggaaggct acccgaaacg tttgacccaa gttaaacaat ttaaaggcaa tgctaccaaa
     3361 tacgcaacaa acttctctct gctgaaacaa gccggagatg tcgaagagaa tcctggaccg
     3421 atggacaaga agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg
     3481 atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg
     3541 cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag
     3601 gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc
     3661 tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga
     3721 ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc
     3781 aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag
     3841 aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac
     3901 atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac
     3961 gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc
     4021 atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga
     4081 cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac
     4141 ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag
     4201 gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc
     4261 cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc
     4321 ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct
     4381 atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg
     4441 cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc
     4501 ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg
     4561 gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg
     4621 aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac
     4681 gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc
     4741 gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc
     4801 agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa
     4861 gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag
     4921 aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg
     4981 tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg
     5041 agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc
     5101 gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc
     5161 tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt
     5221 atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg
     5281 ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc
     5341 cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc
     5401 aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg
     5461 gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac
     5521 agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg
     5581 cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca
     5641 gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg
     5701 atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga
     5761 atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc
     5821 gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg
     5881 gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat
     5941 atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc
     6001 gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag
     6061 aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg
     6121 accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag
     6181 ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac
     6241 actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc
     6301 aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac
     6361 taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag
     6421 taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag
     6481 atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc
     6541 aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg
     6601 cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt
     6661 gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg
     6721 cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc
     6781 gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc
     6841 tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg
     6901 aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac
     6961 tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag
     7021 tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg
     7081 cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc
     7141 cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa
     7201 cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg
     7261 atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag
     7321 cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc
     7381 cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa
     7441 gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc
     7501 gacctgtctc agctgggagg cgacaagcga cctgccgcca caaagaaggc tggacaggct
     7561 aagaagaaga aagattacaa agacgatgac gataagtaac cgcggtctag aacgcgttaa
     7621 gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     7681 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     7741 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     7801 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     7861 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     7921 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     7981 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     8041 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     8101 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     8161 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcgtcga
     8221 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     8281 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
     8341 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     8401 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     8461 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     8521 gtacgtatag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
     8581 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     8641 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     8701 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat
     8761 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     8821 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     8881 cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc
     8941 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     9001 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     9061 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
     9121 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     9181 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     9241 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     9301 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     9361 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     9421 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     9481 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     9541 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     9601 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     9661 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     9721 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     9781 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     9841 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     9901 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     9961 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
    10021 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
    10081 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
    10141 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
    10201 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
    10261 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
    10321 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
    10381 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
    10441 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
    10501 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
    10561 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
    10621 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
    10681 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
    10741 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
    10801 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
    10861 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
    10921 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
    10981 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
    11041 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
    11101 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
    11161 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
    11221 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
    11281 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
    11341 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
    11401 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
    11461 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
    11521 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
    11581 agcgcaacgc aat
//
