LOCUS       sequence-208414-       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        5..231
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             232..412
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    459..584
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1077..1310
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1495..1539
                     /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    1723..1840
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1909..2120
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2140..2149
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2146..5547,5548..5550,5551..5796,5797..5799,5800..6147,
                     6148..6150,6151..6153,6154..6156,6157..6249)
                     /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             6256..6276
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6283..6348
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6349..6363
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             6373..6429
                     /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(6430..6432,6433..6435,6436..7149)
                     /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(6475..6496)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6736..6755)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7083..7104
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        7162..7661
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7197..7220)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7594..7613
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             7682..8281
                     /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(7682..7701)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8178..8198
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8297..8885
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8350..8370)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8768..8779)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8957..9190
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9268..9389
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9305..9324)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9359..9378
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9429..9564
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9584..9603)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9585..9603)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9613..9630)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9613..9629)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9622..9644)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9771..10226
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9858..9877)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10068..10089
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10252..10356
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10357..10425,10426..11217)
                     /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(10575..10594)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11388..11976
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11877..11896
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12130..12147
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12264..12285
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12300..12305,12306..12323,12324..12330)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12338..12354
                     /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     12343..12365
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12362..12378
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12362..12378
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12397..12417
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        12399..12417
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaccc acctcccaac cccgagggga cccagaccac aattttaaaa gaaaaggggg
     1741 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     1801 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     1861 cagagatcca ctttggcgcc ggctcgagtg gctccggtgc ccgtcagtgg gcagagcgca
     1921 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga
     1981 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     2041 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     2101 ggtttgccgc cagaacacag gtgtcgtgac gcgggatccg ccaccatgga taaaaagtat
     2161 tctattggtt tagacatcgg cactaattcc gttggatggg ctgtcataac cgatgaatac
     2221 aaagtacctt caaagaaatt taaggtgttg gggaacacag accgtcattc gattaaaaag
     2281 aatcttatcg gtgccctcct attcgatagt ggcgaaacgg cagaggcgac tcgcctgaaa
     2341 cgaaccgctc ggagaaggta tacacgtcgc aagaaccgaa tatgttactt acaagaaatt
     2401 tttagcaatg agatggccaa agttgacgat tctttctttc accgtttgga agagtccttc
     2461 cttgtcgaag aggacaagaa acatgaacgg caccccatct ttggaaacat agtagatgag
     2521 gtggcatatc atgaaaagta cccaacgatt tatcacctca gaaaaaagct agttgactca
     2581 actgataaag cggacctgag gttaatctac ttggctcttg cccatatgat aaagttccgt
     2641 gggcactttc tcattgaggg tgatctaaat ccggacaact cggatgtcga caaactgttc
     2701 atccagttag tacaaaccta taatcagttg tttgaagaga accctataaa tgcaagtggc
     2761 gtggatgcga aggctattct tagcgcccgc ctctctaaat cccgacggct agaaaacctg
     2821 atcgcacaat tacccggaga gaagaaaaat gggttgttcg gtaaccttat agcgctctca
     2881 ctaggcctga caccaaattt taagtcgaac ttcgacttag ctgaagatgc caaattgcag
     2941 cttagtaagg acacgtacga tgacgatctc gacaatctac tggcacaaat tggagatcag
     3001 tatgcggact tatttttggc tgccaaaaac cttagcgatg caatcctcct atctgacata
     3061 ctgagagtta atactgagat taccaaggcg ccgttatccg cttcaatgat caaaaggtac
     3121 gatgaacatc accaagactt gacacttctc aaggccctag tccgtcagca actgcctgag
     3181 aaatataagg aaatattctt tgatcagtcg aaaaacgggt acgcaggtta tattgacggc
     3241 ggagcgagtc aagaggaatt ctacaagttt atcaaaccca tattagagaa gatggatggg
     3301 acggaagagt tgcttgtaaa actcaatcgc gaagatctac tgcgaaagca gcggactttc
     3361 gacaacggta gcattccaca tcaaatccac ttaggcgaat tgcatgctat acttagaagg
     3421 caggaggatt tttatccgtt cctcaaagac aatcgtgaaa agattgagaa aatcctaacc
     3481 tttcgcatac cttactatgt gggacccctg gcccgaggga actctcggtt cgcatggatg
     3541 acaagaaagt ccgaagaaac gattactcca tggaattttg aggaagttgt cgataaaggt
     3601 gcgtcagctc aatcgttcat cgagaggatg accaactttg acaagaattt accgaacgaa
     3661 aaagtattgc ctaagcacag tttactttac gagtatttca cagtgtacaa tgaactcacg
     3721 aaagttaagt atgtcactga gggcatgcgt aaacccgcct ttctaagcgg agaacagaag
     3781 aaagcaatag tagatctgtt attcaagacc aaccgcaaag tgacagttaa gcaattgaaa
     3841 gaggactact ttaagaaaat tgaatgcttc gattctgtcg agatctccgg ggtagaagat
     3901 cgatttaatg cgtcacttgg tacgtatcat gacctcctaa agataattaa agataaggac
     3961 ttcctggata acgaagagaa tgaagatatc ttagaagata tagtgttgac tcttaccctc
     4021 tttgaagatc gggaaatgat tgaggaaaga ctaaaaacat acgctcacct gttcgacgat
     4081 aaggttatga aacagttaaa gaggcgtcgc tatacgggct ggggacgatt gtcgcggaaa
     4141 cttatcaacg ggataagaga caagcaaagt ggtaaaacta ttctcgattt tctaaagagc
     4201 gacggcttcg ccaataggaa ctttatgcag ctgatccatg atgactcttt aaccttcaaa
     4261 gaggatatac aaaaggcaca ggtttccgga caaggggact cattgcacga acatattgcg
     4321 aatcttgctg gttcgccagc catcaaaaag ggcatactcc agacagtcaa agtagtggat
     4381 gagctagtta aggtcatggg acgtcacaaa ccggaaaaca ttgtaatcga gatggcacgc
     4441 gaaaatcaaa cgactcagaa ggggcaaaaa aacagtcgag agcggatgaa gagaatagaa
     4501 gagggtatta aagaactggg cagccagatc ttaaaggagc atcctgtgga aaatacccaa
     4561 ttgcagaacg agaaacttta cctctattac ctacaaaatg gaagggacat gtatgttgat
     4621 caggaactgg acataaaccg tttatctgat tacgacgtcg atcacattgt accccaatcc
     4681 tttttgaagg acgattcaat cgacaataaa gtgcttacac gctcggataa gaaccgaggg
     4741 aaaagtgaca atgttccaag cgaggaagtc gtaaagaaaa tgaagaacta ttggcggcag
     4801 ctcctaaatg cgaaactgat aacgcaaaga aagttcgata acttaactaa agctgagagg
     4861 ggtggcttgt ctgaacttga caaggccgga tttattaaac gtcagctcgt ggaaacccgc
     4921 caaatcacaa agcatgttgc acagatacta gattcccgaa tgaatacgaa atacgacgag
     4981 aacgataagc tgattcggga agtcaaagta atcactttaa agtcaaaatt ggtgtcggac
     5041 ttcagaaagg attttcaatt ctataaagtt agggagataa ataactacca ccatgcgcac
     5101 gacgcttatc ttaatgccgt cgtagggacc gcactcatta agaaataccc gaagctagaa
     5161 agtgagtttg tgtatggtga ttacaaagtt tatgacgtcc gtaagatgat cgcgaaaagc
     5221 gaacaggaga taggcaaggc tacagccaaa tacttctttt attctaacat tatgaatttc
     5281 tttaagacgg aaatcactct ggcaaacgga gagatacgca aacgaccttt aattgaaacc
     5341 aatggggaga caggtgaaat cgtatgggat aagggccggg acttcgcgac ggtgagaaaa
     5401 gttttgtcca tgccccaagt caacatagta aagaaaactg aggtgcagac cggagggttt
     5461 tcaaaggaat cgattcttcc aaaaaggaat agtgataagc tcatcgctcg taaaaaggac
     5521 tgggacccga aaaagtacgg tggcttcgtg agccctacag ttgcctattc tgtcctagta
     5581 gtggcaaaag ttgagaaggg aaaatccaag aaactgaagt cagtcaaaga attattgggg
     5641 ataacgatta tggagcgctc gtcttttgaa aagaacccca tcgacttcct tgaggcgaaa
     5701 ggttacaagg aagtaaaaaa ggatctcata attaaactac caaagtatag tctgtttgag
     5761 ttagaaaatg gccgaaaacg gatgttggct agcgccagag agcttcaaaa ggggaacgaa
     5821 ctcgcactac cgtctaaata cgtgaatttc ctgtatttag cgtcccatta cgagaagttg
     5881 aaaggttcac ctgaagataa cgaacagaag caactttttg ttgagcagca caaacattat
     5941 ctcgacgaaa tcatagagca aatttcggaa ttcagtaaga gagtcatcct agctgatgcc
     6001 aatctggaca aagtattaag cgcatacaac aagcacaggg ataaacccat acgtgagcag
     6061 gcggaaaata ttatccattt gtttactctt accaacctcg gcgctccagc cgcattcaag
     6121 tattttgaca caacgataga tcgcaaagag tacagatcta ccaaggaggt gctagacgcg
     6181 acactgattc accaatccat cacgggatta tatgaaactc ggatagattt gtcacagctt
     6241 gggggtgacg gatcccccaa gaagaagagg aaagtctcga gcgactacaa agaccatgac
     6301 ggtgattata aagatcatga catcgattac aaggatgacg atgacaagga tgacgatgac
     6361 aagggcagcg gcgccaccaa cttcagcctg ctgaagcagg ccggcgacgt ggaggagaac
     6421 cccggcccca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     6481 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ctggcgaggg cgagggcgat
     6541 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     6601 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     6661 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     6721 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     6781 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     6841 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     6901 cagaagaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga cggcagcgtg
     6961 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     7021 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     7081 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     7141 tacaagtaag atgcatctac cgggtagggg aggcgctttt cccaaggcag tctggagcat
     7201 gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac
     7261 attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac
     7321 cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga
     7381 cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca gatggacagc accgctgagc
     7441 aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct
     7501 gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg
     7561 gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt
     7621 ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcagc ccaagcttac
     7681 catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt
     7741 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga
     7801 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     7861 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     7921 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     7981 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     8041 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     8101 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     8161 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     8221 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     8281 attctagaaa gtcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     8341 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     8401 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     8461 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     8521 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     8581 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     8641 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     8701 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     8761 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     8821 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     8881 cccgcgtcga ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     8941 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt
     9001 gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     9061 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     9121 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     9181 atctctagca gtacgtatag tagttcatgt catcttatta ttcagtattt ataacttgca
     9241 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     9301 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     9361 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     9421 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     9481 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     9541 agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta
     9601 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     9661 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     9721 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     9781 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     9841 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     9901 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     9961 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
    10021 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
    10081 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
    10141 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
    10201 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
    10261 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
    10321 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
    10381 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
    10441 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
    10501 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
    10561 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
    10621 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
    10681 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
    10741 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
    10801 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
    10861 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
    10921 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
    10981 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
    11041 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
    11101 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
    11161 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
    11221 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
    11281 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
    11341 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
    11401 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
    11461 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
    11521 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
    11581 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
    11641 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
    11701 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
    11761 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
    11821 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
    11881 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
    11941 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
    12001 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
    12061 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
    12121 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
    12181 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
    12241 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
    12301 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
    12361 acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa
    12421 caaaagctgg agctgcaagc
//
