LOCUS       sequence-211426-       11203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for constitutive expression of Cas9-VRERRA in 
            mammalian cells (codon optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11203)
  AUTHORS   Zafra MP, Schatoff EM, Katti A, Foronda M, Breinig M, Schweitzer AY,
            Simon A, Han T, Goswami S, Montgomery E, Thibado J, Kastenhuber ER, 
            Sanchez-Rivera FJ, Shi J, Vakoc CR, Lowe SW, Tschaharganeh DF, Dow 
            LE
  TITLE     Optimized base editors enable efficient editing in cells, organoids 
            and mice.
  JOURNAL   Nat Biotechnol. 2018 Jul 3. pii: nbt.4194. doi: 10.1038/nbt.4194.
  PUBMED    29969439
REFERENCE   2  (bases 1 to 11203)
  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: "Nat 
            Biotechnol. 2018 Jul 3. pii: nbt.4194. doi: 10.1038/nbt.4194."
COMMENT     Sequence Label: pLenti-VRERRA-P2A-Puro
FEATURES             Location/Qualifiers
     source          1..11203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             227..407
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    454..579
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1072..1305
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1490..1534
                     /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    1718..1835
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1904..2115
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2135..2144
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2144..2167
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2174..2194
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2219..5617,5618..5620,5621..5866,5867..5869,5870..6217,
                     6218..6220,6221..6223,6224..6226,6227..6319)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/G1218R/R1335E/T1337R mutations"
                     /label=Cas9 VRER
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKEYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             2219..6319
                     /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
                     SILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKEYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6320..6367
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6383..6439
                     /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             6440..7039
                     /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(6440..6459)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6936..6956
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7055..7643
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7108..7128)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7526..7537)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7715..7948
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8026..8147
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8063..8082)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8117..8136
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8187..8322
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8342..8361)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8343..8361)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8371..8388)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8371..8387)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8380..8402)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8529..8984
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8616..8635)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8826..8847
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9010..9114
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9115..9183,9184..9975)
                     /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(9333..9352)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10146..10734
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10635..10654
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10888..10905
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11022..11043
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11058..11063,11064..11081,11082..11088)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11096..11112
                     /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     11101..11123
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11120..11136
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11120..11136
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11155..11175
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11157..11175
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        join(11203,1..226)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc
       61 cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg
      121 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc
      181 gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgggt ctctctggtt
      241 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
      301 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
      361 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac
      421 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct
      481 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact
      541 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt
      601 agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa
      661 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa
      721 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca
      781 gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata
      841 gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag
      901 accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa
      961 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc
     1021 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt
     1081 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac
     1141 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc
     1201 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc
     1261 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg
     1321 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc
     1381 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta
     1441 cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca
     1501 agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg
     1561 gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt
     1621 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca
     1681 gacccacctc ccaaccccga ggggacccag accacaattt taaaagaaaa ggggggattg
     1741 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
     1801 aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag
     1861 atccactttg gcgccggctc gagtggctcc ggtgcccgtc agtgggcaga gcgcacatcg
     1921 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg
     1981 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     2041 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     2101 gccgccagaa cacaggtgtc gtgacgcggg atccgccacc atggattaca aagacgatga
     2161 cgataagatg gccccaaaga agaagcggaa ggtcggtatc cacggagtcc cagcagccga
     2221 caagaagtac agcatcggcc tggacatcgg caccaactct gtgggctggg ccgtgatcac
     2281 cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag
     2341 catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac
     2401 ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct
     2461 gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga
     2521 agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat
     2581 cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact
     2641 ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat
     2701 caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga
     2761 caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa
     2821 cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct
     2881 ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat
     2941 tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc
     3001 caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat
     3061 cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct
     3121 gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat
     3181 caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca
     3241 gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta
     3301 cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa
     3361 gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca
     3421 gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat
     3481 tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa
     3541 gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt
     3601 cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt
     3661 ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct
     3721 gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa
     3781 cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg
     3841 cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa
     3901 gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg
     3961 cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa
     4021 ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac
     4081 cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct
     4141 gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct
     4201 gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt
     4261 cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct
     4321 gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga
     4381 gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa
     4441 ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga
     4501 aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa
     4561 gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga
     4621 aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat
     4681 gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt
     4741 gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa
     4801 gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta
     4861 ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa
     4921 ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt
     4981 ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa
     5041 gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct
     5101 ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca
     5161 ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc
     5221 taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat
     5281 cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat
     5341 catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct
     5401 gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac
     5461 cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac
     5521 aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag
     5581 aaagaaggac tgggacccta agaagtacgg cggcttcgtc agccccaccg tggcctattc
     5641 tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga
     5701 gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct
     5761 ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc
     5821 cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccaggg aactgcagaa
     5881 gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta
     5941 tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca
     6001 caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct
     6061 ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat
     6121 cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc
     6181 cgccttcaag tactttgaca ccaccatcga ccggaaggag tacaggagca ccaaagaggt
     6241 gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct
     6301 gtctcagctg ggaggcgaca agcgtcctgc tgctactaag aaagctggtc aagctaagaa
     6361 aaagaaagct agcggcagcg gcgccaccaa cttcagcctg ctgaagcagg ccggcgacgt
     6421 ggaggagaac cccggcccca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
     6481 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     6541 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     6601 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     6661 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     6721 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     6781 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggagtct cgcccgacca
     6841 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     6901 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     6961 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     7021 ccgcaagccc ggtgcctgaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg
     7081 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     7141 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     7201 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     7261 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     7321 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     7381 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     7441 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     7501 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     7561 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     7621 ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta
     7681 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     7741 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     7801 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     7861 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     7921 ttttagtcag tgtggaaaat ctctagcagt acgtatagta gttcatgtca tcttattatt
     7981 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca
     8041 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     8101 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc
     8161 tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     8221 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     8281 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc
     8341 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     8401 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     8461 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     8521 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     8581 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     8641 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     8701 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     8761 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     8821 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     8881 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     8941 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     9001 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     9061 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     9121 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     9181 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     9241 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     9301 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     9361 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     9421 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     9481 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     9541 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     9601 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     9661 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     9721 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     9781 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     9841 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     9901 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     9961 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
    10021 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
    10081 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
    10141 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
    10201 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
    10261 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
    10321 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
    10381 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
    10441 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
    10501 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
    10561 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
    10621 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
    10681 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
    10741 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
    10801 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
    10861 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
    10921 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
    10981 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
    11041 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
    11101 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
    11161 aaccctcact aaagggaaca aaagctggag ctgcaagctt aat
//
