LOCUS       sequence-211465-       12440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for constitutive expression of Cas9-VRER-P2A-GFP 
            (not codon optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12440)
  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 12440)
  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-VRER-P2A-GFP-PGK-Puro
FEATURES             Location/Qualifiers
     source          1..12440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..250
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             251..431
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    478..603
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1096..1329
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1514..1558
                     /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    1742..1859
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1928..2139
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2159..2168
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2165..5566,5567..5569,5570..5815,5816..5818,5819..6166,
                     6167..6169,6170..6172,6173..6175,6176..6268)
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKEYRSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6275..6295
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6302..6367
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6368..6382
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             6392..6448
                     /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             join(6449..6451,6452..6454,6455..7168)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6494..6515)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6755..6774)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7102..7123
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        7181..7680
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7216..7239)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7613..7632
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             7701..8300
                     /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(7701..7720)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8197..8217
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8316..8904
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8369..8389)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8787..8798)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8976..9209
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9287..9408
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9324..9343)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9378..9397
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9448..9583
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9603..9622)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9604..9622)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9632..9649)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9632..9648)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9641..9663)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9790..10245
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9877..9896)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10087..10108
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10271..10375
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10376..10444,10445..11236)
                     /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(10594..10613)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11407..11995
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11896..11915
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12149..12166
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12283..12304
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12319..12324,12325..12342,12343..12349)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12357..12373
                     /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     12362..12384
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12381..12397
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12381..12397
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12416..12436
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        12418..12436
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt cttgcaacat
       61 ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt
      121 ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg
      181 gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt gcctagctcg
      241 atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
      301 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
      361 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
      421 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc
      481 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg
      541 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc
      601 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg
      661 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc
      721 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta
      781 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc
      841 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata
      901 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc
      961 tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa
     1021 aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa
     1081 aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat
     1141 gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca
     1201 gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt
     1261 ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca
     1321 acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg
     1381 gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga
     1441 gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca
     1501 aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg
     1561 gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg
     1621 aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca
     1681 gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccagaccaca
     1741 attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
     1801 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc
     1861 gggtttatta cagggacagc agagatccac tttggcgccg gctcgagtgg ctccggtgcc
     1921 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
     1981 aattgaaccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
     2041 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     2101 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg tgtcgtgacg cgggatccgc
     2161 caccatggat aaaaagtatt ctattggttt agacatcggc actaattccg ttggatgggc
     2221 tgtcataacc gatgaataca aagtaccttc aaagaaattt aaggtgttgg ggaacacaga
     2281 ccgtcattcg attaaaaaga atcttatcgg tgccctccta ttcgatagtg gcgaaacggc
     2341 agaggcgact cgcctgaaac gaaccgctcg gagaaggtat acacgtcgca agaaccgaat
     2401 atgttactta caagaaattt ttagcaatga gatggccaaa gttgacgatt ctttctttca
     2461 ccgtttggaa gagtccttcc ttgtcgaaga ggacaagaaa catgaacggc accccatctt
     2521 tggaaacata gtagatgagg tggcatatca tgaaaagtac ccaacgattt atcacctcag
     2581 aaaaaagcta gttgactcaa ctgataaagc ggacctgagg ttaatctact tggctcttgc
     2641 ccatatgata aagttccgtg ggcactttct cattgagggt gatctaaatc cggacaactc
     2701 ggatgtcgac aaactgttca tccagttagt acaaacctat aatcagttgt ttgaagagaa
     2761 ccctataaat gcaagtggcg tggatgcgaa ggctattctt agcgcccgcc tctctaaatc
     2821 ccgacggcta gaaaacctga tcgcacaatt acccggagag aagaaaaatg ggttgttcgg
     2881 taaccttata gcgctctcac taggcctgac accaaatttt aagtcgaact tcgacttagc
     2941 tgaagatgcc aaattgcagc ttagtaagga cacgtacgat gacgatctcg acaatctact
     3001 ggcacaaatt ggagatcagt atgcggactt atttttggct gccaaaaacc ttagcgatgc
     3061 aatcctccta tctgacatac tgagagttaa tactgagatt accaaggcgc cgttatccgc
     3121 ttcaatgatc aaaaggtacg atgaacatca ccaagacttg acacttctca aggccctagt
     3181 ccgtcagcaa ctgcctgaga aatataagga aatattcttt gatcagtcga aaaacgggta
     3241 cgcaggttat attgacggcg gagcgagtca agaggaattc tacaagttta tcaaacccat
     3301 attagagaag atggatggga cggaagagtt gcttgtaaaa ctcaatcgcg aagatctact
     3361 gcgaaagcag cggactttcg acaacggtag cattccacat caaatccact taggcgaatt
     3421 gcatgctata cttagaaggc aggaggattt ttatccgttc ctcaaagaca atcgtgaaaa
     3481 gattgagaaa atcctaacct ttcgcatacc ttactatgtg ggacccctgg cccgagggaa
     3541 ctctcggttc gcatggatga caagaaagtc cgaagaaacg attactccat ggaattttga
     3601 ggaagttgtc gataaaggtg cgtcagctca atcgttcatc gagaggatga ccaactttga
     3661 caagaattta ccgaacgaaa aagtattgcc taagcacagt ttactttacg agtatttcac
     3721 agtgtacaat gaactcacga aagttaagta tgtcactgag ggcatgcgta aacccgcctt
     3781 tctaagcgga gaacagaaga aagcaatagt agatctgtta ttcaagacca accgcaaagt
     3841 gacagttaag caattgaaag aggactactt taagaaaatt gaatgcttcg attctgtcga
     3901 gatctccggg gtagaagatc gatttaatgc gtcacttggt acgtatcatg acctcctaaa
     3961 gataattaaa gataaggact tcctggataa cgaagagaat gaagatatct tagaagatat
     4021 agtgttgact cttaccctct ttgaagatcg ggaaatgatt gaggaaagac taaaaacata
     4081 cgctcacctg ttcgacgata aggttatgaa acagttaaag aggcgtcgct atacgggctg
     4141 gggacgattg tcgcggaaac ttatcaacgg gataagagac aagcaaagtg gtaaaactat
     4201 tctcgatttt ctaaagagcg acggcttcgc caataggaac tttatgcagc tgatccatga
     4261 tgactcttta accttcaaag aggatataca aaaggcacag gtttccggac aaggggactc
     4321 attgcacgaa catattgcga atcttgctgg ttcgccagcc atcaaaaagg gcatactcca
     4381 gacagtcaaa gtagtggatg agctagttaa ggtcatggga cgtcacaaac cggaaaacat
     4441 tgtaatcgag atggcacgcg aaaatcaaac gactcagaag gggcaaaaaa acagtcgaga
     4501 gcggatgaag agaatagaag agggtattaa agaactgggc agccagatct taaaggagca
     4561 tcctgtggaa aatacccaat tgcagaacga gaaactttac ctctattacc tacaaaatgg
     4621 aagggacatg tatgttgatc aggaactgga cataaaccgt ttatctgatt acgacgtcga
     4681 tcacattgta ccccaatcct ttttgaagga cgattcaatc gacaataaag tgcttacacg
     4741 ctcggataag aaccgaggga aaagtgacaa tgttccaagc gaggaagtcg taaagaaaat
     4801 gaagaactat tggcggcagc tcctaaatgc gaaactgata acgcaaagaa agttcgataa
     4861 cttaactaaa gctgagaggg gtggcttgtc tgaacttgac aaggccggat ttattaaacg
     4921 tcagctcgtg gaaacccgcc aaatcacaaa gcatgttgca cagatactag attcccgaat
     4981 gaatacgaaa tacgacgaga acgataagct gattcgggaa gtcaaagtaa tcactttaaa
     5041 gtcaaaattg gtgtcggact tcagaaagga ttttcaattc tataaagtta gggagataaa
     5101 taactaccac catgcgcacg acgcttatct taatgccgtc gtagggaccg cactcattaa
     5161 gaaatacccg aagctagaaa gtgagtttgt gtatggtgat tacaaagttt atgacgtccg
     5221 taagatgatc gcgaaaagcg aacaggagat aggcaaggct acagccaaat acttctttta
     5281 ttctaacatt atgaatttct ttaagacgga aatcactctg gcaaacggag agatacgcaa
     5341 acgaccttta attgaaacca atggggagac aggtgaaatc gtatgggata agggccggga
     5401 cttcgcgacg gtgagaaaag ttttgtccat gccccaagtc aacatagtaa agaaaactga
     5461 ggtgcagacc ggagggtttt caaaggaatc gattcttcca aaaaggaata gtgataagct
     5521 catcgctcgt aaaaaggact gggacccgaa aaagtacggt ggcttcgtga gccctacagt
     5581 tgcctattct gtcctagtag tggcaaaagt tgagaaggga aaatccaaga aactgaagtc
     5641 agtcaaagaa ttattgggga taacgattat ggagcgctcg tcttttgaaa agaaccccat
     5701 cgacttcctt gaggcgaaag gttacaagga agtaaaaaag gatctcataa ttaaactacc
     5761 aaagtatagt ctgtttgagt tagaaaatgg ccgaaaacgg atgttggcta gcgccagaga
     5821 gcttcaaaag gggaacgaac tcgcactacc gtctaaatac gtgaatttcc tgtatttagc
     5881 gtcccattac gagaagttga aaggttcacc tgaagataac gaacagaagc aactttttgt
     5941 tgagcagcac aaacattatc tcgacgaaat catagagcaa atttcggaat tcagtaagag
     6001 agtcatccta gctgatgcca atctggacaa agtattaagc gcatacaaca agcacaggga
     6061 taaacccata cgtgagcagg cggaaaatat tatccatttg tttactctta ccaacctcgg
     6121 cgctccagcc gcattcaagt attttgacac aacgatagat cgcaaagagt acagatctac
     6181 caaggaggtg ctagacgcga cactgattca ccaatccatc acgggattat atgaaactcg
     6241 gatagatttg tcacagcttg ggggtgacgg atcccccaag aagaagagga aagtctcgag
     6301 cgactacaaa gaccatgacg gtgattataa agatcatgac atcgattaca aggatgacga
     6361 tgacaaggat gacgatgaca agggcagcgg cgccaccaac ttcagcctgc tgaagcaggc
     6421 cggcgacgtg gaggagaacc ccggccccat ggtgagcaag ggcgaggagc tgttcaccgg
     6481 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     6541 tggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     6601 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     6661 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     6721 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     6781 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     6841 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     6901 ctatatcatg gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa
     6961 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     7021 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     7081 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     7141 tctcggcatg gacgagctgt acaagtaaga tgcatctacc gggtagggga ggcgcttttc
     7201 ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt
     7261 ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt
     7321 ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg
     7381 cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag
     7441 atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag
     7501 ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc
     7561 aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc
     7621 acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg
     7681 acctgcagcc caagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     7741 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     7801 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     7861 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     7921 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     7981 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     8041 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     8101 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     8161 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     8221 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     8281 cccgcaagcc cggtgcctga ttctagaaag tcgacaatca acctctggat tacaaaattt
     8341 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     8401 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     8461 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     8521 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     8581 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     8641 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     8701 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     8761 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     8821 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     8881 tctccctttg ggccgcctcc ccgcgtcgac tttaagacca atgacttaca aggcagctgt
     8941 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     9001 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     9061 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     9121 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     9181 cttttagtca gtgtggaaaa tctctagcag tacgtatagt agttcatgtc atcttattat
     9241 tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc
     9301 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     9361 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct
     9421 ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     9481 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     9541 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt
     9601 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
     9661 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     9721 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     9781 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     9841 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     9901 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     9961 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
    10021 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
    10081 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
    10141 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
    10201 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
    10261 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
    10321 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
    10381 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
    10441 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
    10501 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
    10561 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
    10621 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
    10681 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
    10741 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
    10801 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
    10861 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
    10921 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
    10981 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
    11041 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
    11101 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
    11161 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
    11221 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
    11281 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
    11341 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
    11401 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
    11461 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
    11521 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
    11581 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
    11641 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
    11701 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
    11761 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
    11821 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
    11881 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
    11941 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
    12001 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
    12061 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
    12121 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
    12181 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
    12241 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
    12301 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
    12361 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat
    12421 taaccctcac taaagggaac
//
