LOCUS       sequence-211467-       12425 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for constitutive expression of Cas9-HF1-P2A-GFP 
            (not codon optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12425)
  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 12425)
  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-HF1-P2A-GFP-PGK-Puro
FEATURES             Location/Qualifiers
     source          1..12425
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..229
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1075..1308
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /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    1721..1838
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1907..2118
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2138..2147
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2144..3631,3632..3634,3635..4123,4124..4126,4127..4225,
                     4226..4228,4229..4918,4919..4921,4922..6247)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity (Kleinstiver et al., 2016)"
                     /label=SpCas9-HF1
                     /note="carries the mutations N497A, R661A, Q695A, and 
                     Q926A"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTAFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGALSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     ALIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRAITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6254..6274
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6281..6346
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6356..6412
                     /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(6413..6415,6416..6418,6419..7132)
                     /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(6458..6479)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6719..6738)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7066..7087
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        7145..7644
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7180..7203)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7577..7596
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             7665..8264
                     /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(7665..7684)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8161..8181
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8280..8868
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8333..8353)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8751..8762)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8940..9173
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9251..9372
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9288..9307)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9342..9361
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9412..9547
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9567..9586)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9568..9586)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9596..9613)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9596..9612)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9605..9627)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9754..10209
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9841..9860)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10051..10072
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10235..10339
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10340..10408,10409..11200)
                     /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(10558..10577)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11371..11959
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11860..11879
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12113..12130
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12247..12268
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12283..12288,12289..12306,12307..12313)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12321..12337
                     /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     12326..12348
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12345..12361
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12345..12361
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12380..12400
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        12382..12400
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cagaccacaa ttttaaaaga aaagggggga
     1741 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     1801 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
     1861 gagatccact ttggcgccgg ctcgagtggc tccggtgccc gtcagtgggc agagcgcaca
     1921 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga
     1981 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     2041 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     2101 tttgccgcca gaacacaggt gtcgtgacgc gggatccgcc accatggata aaaagtattc
     2161 tattggttta gacatcggca ctaattccgt tggatgggct gtcataaccg atgaatacaa
     2221 agtaccttca aagaaattta aggtgttggg gaacacagac cgtcattcga ttaaaaagaa
     2281 tcttatcggt gccctcctat tcgatagtgg cgaaacggca gaggcgactc gcctgaaacg
     2341 aaccgctcgg agaaggtata cacgtcgcaa gaaccgaata tgttacttac aagaaatttt
     2401 tagcaatgag atggccaaag ttgacgattc tttctttcac cgtttggaag agtccttcct
     2461 tgtcgaagag gacaagaaac atgaacggca ccccatcttt ggaaacatag tagatgaggt
     2521 ggcatatcat gaaaagtacc caacgattta tcacctcaga aaaaagctag ttgactcaac
     2581 tgataaagcg gacctgaggt taatctactt ggctcttgcc catatgataa agttccgtgg
     2641 gcactttctc attgagggtg atctaaatcc ggacaactcg gatgtcgaca aactgttcat
     2701 ccagttagta caaacctata atcagttgtt tgaagagaac cctataaatg caagtggcgt
     2761 ggatgcgaag gctattctta gcgcccgcct ctctaaatcc cgacggctag aaaacctgat
     2821 cgcacaatta cccggagaga agaaaaatgg gttgttcggt aaccttatag cgctctcact
     2881 aggcctgaca ccaaatttta agtcgaactt cgacttagct gaagatgcca aattgcagct
     2941 tagtaaggac acgtacgatg acgatctcga caatctactg gcacaaattg gagatcagta
     3001 tgcggactta tttttggctg ccaaaaacct tagcgatgca atcctcctat ctgacatact
     3061 gagagttaat actgagatta ccaaggcgcc gttatccgct tcaatgatca aaaggtacga
     3121 tgaacatcac caagacttga cacttctcaa ggccctagtc cgtcagcaac tgcctgagaa
     3181 atataaggaa atattctttg atcagtcgaa aaacgggtac gcaggttata ttgacggcgg
     3241 agcgagtcaa gaggaattct acaagtttat caaacccata ttagagaaga tggatgggac
     3301 ggaagagttg cttgtaaaac tcaatcgcga agatctactg cgaaagcagc ggactttcga
     3361 caacggtagc attccacatc aaatccactt aggcgaattg catgctatac ttagaaggca
     3421 ggaggatttt tatccgttcc tcaaagacaa tcgtgaaaag attgagaaaa tcctaacctt
     3481 tcgcatacct tactatgtgg gacccctggc ccgagggaac tctcggttcg catggatgac
     3541 aagaaagtcc gaagaaacga ttactccctg gaattttgag gaagttgtcg ataaaggtgc
     3601 gtcagctcaa tcgttcatcg agaggatgac cgcctttgac aagaatttac cgaacgaaaa
     3661 agtattgcct aagcacagtt tactttacga gtatttcaca gtgtacaatg aactcacgaa
     3721 agttaagtat gtcactgagg gcatgcgtaa acccgccttt ctaagcggag aacagaagaa
     3781 agcaatagta gatctgttat tcaagaccaa ccgcaaagtg acagttaagc aattgaaaga
     3841 ggactacttt aagaaaattg aatgcttcga ttctgtcgag atctccgggg tagaagatcg
     3901 atttaatgcg tcacttggta cgtatcatga cctcctaaag ataattaaag ataaggactt
     3961 cctggataac gaagagaatg aagatatctt agaagatata gtgttgactc ttaccctctt
     4021 tgaagatcgg gaaatgattg aggaaagact aaaaacatac gctcacctgt tcgacgataa
     4081 ggttatgaaa cagttaaaga ggcgtcgcta tacgggctgg ggagccttgt cgcggaaact
     4141 tatcaacggg ataagagaca agcaaagtgg taaaactatt ctcgattttc taaagagcga
     4201 cggcttcgcc aataggaact ttatggccct gatccatgat gactctttaa ccttcaaaga
     4261 ggatatacaa aaggcacagg tttccggaca aggggactca ttgcacgaac atattgcgaa
     4321 tcttgctggt tcgccagcca tcaaaaaggg catactccag acagtcaaag tagtggatga
     4381 gctagttaag gtcatgggac gtcacaaacc ggaaaacatt gtaatcgaga tggcacgcga
     4441 aaatcaaacg actcagaagg ggcaaaaaaa cagtcgagag cggatgaaga gaatagaaga
     4501 gggtattaaa gaactgggca gccagatctt aaaggagcat cctgtggaaa atacccaatt
     4561 gcagaacgag aaactttacc tctattacct acaaaatgga agggacatgt atgttgatca
     4621 ggaactggac ataaaccgtt tatctgatta cgacgtcgat cacattgtac cccaatcctt
     4681 tttgaaggac gattcaatcg acaataaagt gcttacacgc tcggataaga accgagggaa
     4741 aagtgacaat gttccaagcg aggaagtcgt aaagaaaatg aagaactatt ggcggcagct
     4801 cctaaatgcg aaactgataa cgcaaagaaa gttcgataac ttaactaaag ctgagagggg
     4861 tggcttgtct gaacttgaca aggccggatt tattaaacgt cagctcgtgg aaacccgcgc
     4921 catcacaaag catgttgcgc agatactaga ttcccgaatg aatacgaaat acgacgagaa
     4981 cgataagctg attcgggaag tcaaagtaat cactttaaag tcaaaattgg tgtcggactt
     5041 cagaaaggat tttcaattct ataaagttag ggagataaat aactaccacc atgcgcacga
     5101 cgcttatctt aatgccgtcg tagggaccgc actcattaag aaatacccga agctagaaag
     5161 tgagtttgtg tatggtgatt acaaagttta tgacgtccgt aagatgatcg cgaaaagcga
     5221 acaggagata ggcaaggcta cagccaaata cttcttttat tctaacatta tgaatttctt
     5281 taagacggaa atcactctgg caaacggaga gatacgcaaa cgacctttaa ttgaaaccaa
     5341 tggggagaca ggtgaaatcg tatgggataa gggccgggac ttcgcgacgg tgagaaaagt
     5401 tttgtccatg ccccaagtca acatagtaaa gaaaactgag gtgcagaccg gagggttttc
     5461 aaaggaatcg attcttccaa aaaggaatag tgataagctc atcgctcgta aaaaggactg
     5521 ggacccgaaa aagtacggtg gcttcgatag ccctacagtt gcctattctg tcctagtagt
     5581 ggcaaaagtt gagaagggaa aatccaagaa actgaagtca gtcaaagaat tattggggat
     5641 aacgattatg gagcgctcgt cttttgaaaa gaaccccatc gacttccttg aggcgaaagg
     5701 ttacaaggaa gtaaaaaagg atctcataat taaactacca aagtatagtc tgtttgagtt
     5761 agaaaatggc cgaaaacgga tgttggctag cgccggagag cttcaaaagg ggaacgaact
     5821 cgcactaccg tctaaatacg tgaatttcct gtatttagcg tcccattacg agaagttgaa
     5881 aggttcacct gaagataacg aacagaagca actttttgtt gagcagcaca aacattatct
     5941 cgacgaaatc atagagcaaa tttcggaatt cagtaagaga gtcatcctag ctgatgccaa
     6001 tctggacaaa gtattaagcg catacaacaa gcacagggat aaacccatac gtgagcaggc
     6061 ggaaaatatt atccatttgt ttactcttac caacctcggc gctccagccg cattcaagta
     6121 ttttgacaca acgatagatc gcaaacgata cacttctacc aaggaggtgc tagacgcgac
     6181 actgattcac caatccatca cgggattata tgaaactcgg atagatttgt cacagcttgg
     6241 gggtgacgga tcccccaaga agaagaggaa agtctcgagc gactacaaag accatgacgg
     6301 tgattataaa gatcatgaca tcgattacaa ggatgacgat gacaagggca gcggcgccac
     6361 caacttcagc ctgctgaagc aggccggcga cgtggaggag aaccccggcc ccatggtgag
     6421 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     6481 aaacggccac aagttcagcg tgtctggcga gggcgagggc gatgccacct acggcaagct
     6541 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     6601 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     6661 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     6721 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     6781 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     6841 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     6901 ggcgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     6961 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     7021 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     7081 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aagatgcatc
     7141 taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg
     7201 ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt
     7261 aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta
     7321 gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta
     7381 gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc
     7441 ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga
     7501 aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg
     7561 tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc
     7621 tcttcctcat ctccgggcct ttcgacctgc agcccaagct taccatgacc gagtacaagc
     7681 ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg
     7741 cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg
     7801 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     7861 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     7921 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     7981 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     8041 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     8101 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     8161 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     8221 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgattctag aaagtcgaca
     8281 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     8341 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     8401 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     8461 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     8521 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     8581 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     8641 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     8701 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     8761 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     8821 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcgt cgactttaag
     8881 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
     8941 ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct
     9001 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     9061 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     9121 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtacgta
     9181 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     9241 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     9301 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     9361 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc
     9421 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     9481 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     9541 ggaggcctag ggacgtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg
     9601 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     9661 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     9721 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     9781 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     9841 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     9901 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     9961 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
    10021 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
    10081 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
    10141 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
    10201 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
    10261 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
    10321 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
    10381 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
    10441 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
    10501 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
    10561 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
    10621 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
    10681 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
    10741 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
    10801 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
    10861 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
    10921 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
    10981 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
    11041 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
    11101 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
    11161 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
    11221 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
    11281 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
    11341 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
    11401 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
    11461 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
    11521 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
    11581 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
    11641 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
    11701 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
    11761 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
    11821 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
    11881 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
    11941 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
    12001 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
    12061 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
    12121 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
    12181 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
    12241 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
    12301 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
    12361 ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc tggagctgca
    12421 agctt
//
