LOCUS       sequence-211460-       11642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for constitutive expression of Cas9-HF1 (not codon
            optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11642)
  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 11642)
  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-PGK-Puro
FEATURES             Location/Qualifiers
     source          1..11642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..233
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             234..414
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    461..586
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1079..1312
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1497..1541
                     /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    1725..1842
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1911..2122
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2142..2151
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2148..3635,3636..3638,3639..4127,4128..4130,4131..4229,
                     4230..4232,4233..4922,4923..4925,4926..6251)
                     /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             6258..6278
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6285..6350
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        6366..6865
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6401..6424)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6798..6817
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6886..7485
                     /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(6886..6905)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7382..7402
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7501..8089
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7554..7574)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7972..7983)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8161..8394
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8472..8593
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8509..8528)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8563..8582
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8633..8768
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8788..8807)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8789..8807)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8817..8834)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8817..8833)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8826..8848)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8975..9430
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9062..9081)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9272..9293
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9456..9560
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9561..9629,9630..10421)
                     /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(9779..9798)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10592..11180
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11081..11100
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11334..11351
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11468..11489
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11504..11509,11510..11527,11528..11534)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11542..11558
                     /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     11547..11569
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11566..11582
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11566..11582
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11601..11621
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11603..11621
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gcttaatgta gtcttatgca atactcttgt agtcttgcaa catggtaacg atgagttagc
       61 aacatgcctt acaaggagag aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt
      121 acgatcgtgc cttattagga aggcaacaga cgggtctgac atggattgga cgaaccactg
      181 aattgccgca ttgcagagat attgtattta agtgcctagc tcgatacata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
      481 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
      541 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
      601 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
      661 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
      721 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
      781 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
      841 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
      901 aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga
      961 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag
     1021 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag
     1081 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga
     1141 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc
     1201 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc
     1261 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt
     1321 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta
     1381 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta
     1441 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga
     1501 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa
     1561 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa
     1621 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat
     1681 cgtttcagac ccacctccca accccgaggg gacccagacc acaattttaa aagaaaaggg
     1741 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     1801 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
     1861 agcagagatc cactttggcg ccggctcgag tggctccggt gcccgtcagt gggcagagcg
     1921 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta
     1981 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
     2041 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
     2101 cgggtttgcc gccagaacac aggtgtcgtg acgcgggatc cgccaccatg gataaaaagt
     2161 attctattgg tttagacatc ggcactaatt ccgttggatg ggctgtcata accgatgaat
     2221 acaaagtacc ttcaaagaaa tttaaggtgt tggggaacac agaccgtcat tcgattaaaa
     2281 agaatcttat cggtgccctc ctattcgata gtggcgaaac ggcagaggcg actcgcctga
     2341 aacgaaccgc tcggagaagg tatacacgtc gcaagaaccg aatatgttac ttacaagaaa
     2401 tttttagcaa tgagatggcc aaagttgacg attctttctt tcaccgtttg gaagagtcct
     2461 tccttgtcga agaggacaag aaacatgaac ggcaccccat ctttggaaac atagtagatg
     2521 aggtggcata tcatgaaaag tacccaacga tttatcacct cagaaaaaag ctagttgact
     2581 caactgataa agcggacctg aggttaatct acttggctct tgcccatatg ataaagttcc
     2641 gtgggcactt tctcattgag ggtgatctaa atccggacaa ctcggatgtc gacaaactgt
     2701 tcatccagtt agtacaaacc tataatcagt tgtttgaaga gaaccctata aatgcaagtg
     2761 gcgtggatgc gaaggctatt cttagcgccc gcctctctaa atcccgacgg ctagaaaacc
     2821 tgatcgcaca attacccgga gagaagaaaa atgggttgtt cggtaacctt atagcgctct
     2881 cactaggcct gacaccaaat tttaagtcga acttcgactt agctgaagat gccaaattgc
     2941 agcttagtaa ggacacgtac gatgacgatc tcgacaatct actggcacaa attggagatc
     3001 agtatgcgga cttatttttg gctgccaaaa accttagcga tgcaatcctc ctatctgaca
     3061 tactgagagt taatactgag attaccaagg cgccgttatc cgcttcaatg atcaaaaggt
     3121 acgatgaaca tcaccaagac ttgacacttc tcaaggccct agtccgtcag caactgcctg
     3181 agaaatataa ggaaatattc tttgatcagt cgaaaaacgg gtacgcaggt tatattgacg
     3241 gcggagcgag tcaagaggaa ttctacaagt ttatcaaacc catattagag aagatggatg
     3301 ggacggaaga gttgcttgta aaactcaatc gcgaagatct actgcgaaag cagcggactt
     3361 tcgacaacgg tagcattcca catcaaatcc acttaggcga attgcatgct atacttagaa
     3421 ggcaggagga tttttatccg ttcctcaaag acaatcgtga aaagattgag aaaatcctaa
     3481 cctttcgcat accttactat gtgggacccc tggcccgagg gaactctcgg ttcgcatgga
     3541 tgacaagaaa gtccgaagaa acgattactc cctggaattt tgaggaagtt gtcgataaag
     3601 gtgcgtcagc tcaatcgttc atcgagagga tgaccgcctt tgacaagaat ttaccgaacg
     3661 aaaaagtatt gcctaagcac agtttacttt acgagtattt cacagtgtac aatgaactca
     3721 cgaaagttaa gtatgtcact gagggcatgc gtaaacccgc ctttctaagc ggagaacaga
     3781 agaaagcaat agtagatctg ttattcaaga ccaaccgcaa agtgacagtt aagcaattga
     3841 aagaggacta ctttaagaaa attgaatgct tcgattctgt cgagatctcc ggggtagaag
     3901 atcgatttaa tgcgtcactt ggtacgtatc atgacctcct aaagataatt aaagataagg
     3961 acttcctgga taacgaagag aatgaagata tcttagaaga tatagtgttg actcttaccc
     4021 tctttgaaga tcgggaaatg attgaggaaa gactaaaaac atacgctcac ctgttcgacg
     4081 ataaggttat gaaacagtta aagaggcgtc gctatacggg ctggggagcc ttgtcgcgga
     4141 aacttatcaa cgggataaga gacaagcaaa gtggtaaaac tattctcgat tttctaaaga
     4201 gcgacggctt cgccaatagg aactttatgg ccctgatcca tgatgactct ttaaccttca
     4261 aagaggatat acaaaaggca caggtttccg gacaagggga ctcattgcac gaacatattg
     4321 cgaatcttgc tggttcgcca gccatcaaaa agggcatact ccagacagtc aaagtagtgg
     4381 atgagctagt taaggtcatg ggacgtcaca aaccggaaaa cattgtaatc gagatggcac
     4441 gcgaaaatca aacgactcag aaggggcaaa aaaacagtcg agagcggatg aagagaatag
     4501 aagagggtat taaagaactg ggcagccaga tcttaaagga gcatcctgtg gaaaataccc
     4561 aattgcagaa cgagaaactt tacctctatt acctacaaaa tggaagggac atgtatgttg
     4621 atcaggaact ggacataaac cgtttatctg attacgacgt cgatcacatt gtaccccaat
     4681 cctttttgaa ggacgattca atcgacaata aagtgcttac acgctcggat aagaaccgag
     4741 ggaaaagtga caatgttcca agcgaggaag tcgtaaagaa aatgaagaac tattggcggc
     4801 agctcctaaa tgcgaaactg ataacgcaaa gaaagttcga taacttaact aaagctgaga
     4861 ggggtggctt gtctgaactt gacaaggccg gatttattaa acgtcagctc gtggaaaccc
     4921 gcgccatcac aaagcatgtt gcgcagatac tagattcccg aatgaatacg aaatacgacg
     4981 agaacgataa gctgattcgg gaagtcaaag taatcacttt aaagtcaaaa ttggtgtcgg
     5041 acttcagaaa ggattttcaa ttctataaag ttagggagat aaataactac caccatgcgc
     5101 acgacgctta tcttaatgcc gtcgtaggga ccgcactcat taagaaatac ccgaagctag
     5161 aaagtgagtt tgtgtatggt gattacaaag tttatgacgt ccgtaagatg atcgcgaaaa
     5221 gcgaacagga gataggcaag gctacagcca aatacttctt ttattctaac attatgaatt
     5281 tctttaagac ggaaatcact ctggcaaacg gagagatacg caaacgacct ttaattgaaa
     5341 ccaatgggga gacaggtgaa atcgtatggg ataagggccg ggacttcgcg acggtgagaa
     5401 aagttttgtc catgccccaa gtcaacatag taaagaaaac tgaggtgcag accggagggt
     5461 tttcaaagga atcgattctt ccaaaaagga atagtgataa gctcatcgct cgtaaaaagg
     5521 actgggaccc gaaaaagtac ggtggcttcg atagccctac agttgcctat tctgtcctag
     5581 tagtggcaaa agttgagaag ggaaaatcca agaaactgaa gtcagtcaaa gaattattgg
     5641 ggataacgat tatggagcgc tcgtcttttg aaaagaaccc catcgacttc cttgaggcga
     5701 aaggttacaa ggaagtaaaa aaggatctca taattaaact accaaagtat agtctgtttg
     5761 agttagaaaa tggccgaaaa cggatgttgg ctagcgccgg agagcttcaa aaggggaacg
     5821 aactcgcact accgtctaaa tacgtgaatt tcctgtattt agcgtcccat tacgagaagt
     5881 tgaaaggttc acctgaagat aacgaacaga agcaactttt tgttgagcag cacaaacatt
     5941 atctcgacga aatcatagag caaatttcgg aattcagtaa gagagtcatc ctagctgatg
     6001 ccaatctgga caaagtatta agcgcataca acaagcacag ggataaaccc atacgtgagc
     6061 aggcggaaaa tattatccat ttgtttactc ttaccaacct cggcgctcca gccgcattca
     6121 agtattttga cacaacgata gatcgcaaac gatacacttc taccaaggag gtgctagacg
     6181 cgacactgat tcaccaatcc atcacgggat tatatgaaac tcggatagat ttgtcacagc
     6241 ttgggggtga cggatccccc aagaagaaga ggaaagtctc gagcgactac aaagaccatg
     6301 acggtgatta taaagatcat gacatcgatt acaaggatga cgatgacaag taagatgcat
     6361 ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc
     6421 gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg
     6481 taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct
     6541 agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt
     6601 agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg
     6661 cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg
     6721 aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag
     6781 gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc
     6841 ctcttcctca tctccgggcc tttcgacctg cagcccaagc ttaccatgac cgagtacaag
     6901 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     6961 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     7021 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     7081 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     7141 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     7201 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     7261 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     7321 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     7381 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     7441 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgattcta gaaagtcgac
     7501 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     7561 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     7621 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     7681 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     7741 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     7801 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     7861 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     7921 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     7981 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     8041 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg tcgactttaa
     8101 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     8161 tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc
     8221 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     8281 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     8341 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtacgt
     8401 atagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     8461 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     8521 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     8581 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccatcccgcc
     8641 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     8701 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     8761 tggaggccta gggacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg
     8821 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     8881 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     8941 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     9001 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     9061 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     9121 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     9181 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     9241 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     9301 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     9361 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     9421 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     9481 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     9541 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     9601 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     9661 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     9721 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     9781 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     9841 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     9901 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     9961 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
    10021 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
    10081 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
    10141 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
    10201 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
    10261 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
    10321 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
    10381 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
    10441 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
    10501 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
    10561 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
    10621 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
    10681 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
    10741 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
    10801 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
    10861 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
    10921 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
    10981 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
    11041 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
    11101 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
    11161 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
    11221 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
    11281 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
    11341 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
    11401 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
    11461 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
    11521 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
    11581 accatgatta cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggagctgc
    11641 aa
//
