LOCUS       sequence-211424-       11188 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 11188)
  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 11188)
  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-Puro
FEATURES             Location/Qualifiers
     source          1..11188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             209..389
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    436..561
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1054..1287
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1472..1516
                     /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    1700..1817
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1886..2097
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2117..2126
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2123..3610,3611..3613,3614..4102,4103..4105,4106..4204,
                     4205..4207,4208..4897,4898..4900,4901..6226)
                     /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             6233..6253
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6260..6325
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6350..6406
                     /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             6407..7006
                     /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(6407..6426)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6903..6923
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7022..7610
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7075..7095)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7493..7504)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7682..7915
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7993..8114
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8030..8049)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8084..8103
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8154..8289
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8309..8328)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8310..8328)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8338..8355)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8338..8354)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8347..8369)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8496..8951
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8583..8602)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8793..8814
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8977..9081
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9082..9150,9151..9942)
                     /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(9300..9319)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10113..10701
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10602..10621
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10855..10872
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10989..11010
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11025..11030,11031..11048,11049..11055)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11063..11079
                     /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     11068..11090
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11087..11103
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11087..11103
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11122..11142
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11124..11142
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        join(11170..11188,1..208)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 cttgtagtct tgcaacatgg taacgatgag ttagcaacat gccttacaag gagagaaaaa
       61 gcaccgtgca tgccgattgg tggaagtaag gtggtacgat cgtgccttat taggaaggca
      121 acagacgggt ctgacatgga ttggacgaac cactgaattg ccgcattgca gagatattgt
      181 atttaagtgc ctagctcgat acataaacgg gtctctctgg ttagaccaga tctgagcctg
      241 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
      301 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
      361 cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt gaaagcgaaa
      421 gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga
      481 ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg ctagaaggag
      541 agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg atgggaaaaa
      601 attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt atgggcaagc
      661 agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga aggctgtaga
      721 caaatactgg gacagctaca accatccctt cagacaggat cagaagaact tagatcatta
      781 tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa agacaccaag
      841 gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc acagcaagcg
      901 gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa gtgaattata
      961 taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg caaagagaag
     1021 agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg ggttcttggg
     1081 agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg ccagacaatt
     1141 attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg cgcaacagca
     1201 tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc tggctgtgga
     1261 aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa aactcatttg
     1321 caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac agatttggaa
     1381 tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct taatacactc
     1441 cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat tggaattaga
     1501 taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt atataaaatt
     1561 attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg tactttctat
     1621 agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc tcccaacccc
     1681 gaggggaccc agaccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg
     1741 aaagaatagt agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta
     1801 caaaaattca aaattttcgg gtttattaca gggacagcag agatccactt tggcgccggc
     1861 tcgagtggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     1921 ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg
     1981 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     2041 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggtg
     2101 tcgtgacgcg ggatccgcca ccatggataa aaagtattct attggtttag acatcggcac
     2161 taattccgtt ggatgggctg tcataaccga tgaatacaaa gtaccttcaa agaaatttaa
     2221 ggtgttgggg aacacagacc gtcattcgat taaaaagaat cttatcggtg ccctcctatt
     2281 cgatagtggc gaaacggcag aggcgactcg cctgaaacga accgctcgga gaaggtatac
     2341 acgtcgcaag aaccgaatat gttacttaca agaaattttt agcaatgaga tggccaaagt
     2401 tgacgattct ttctttcacc gtttggaaga gtccttcctt gtcgaagagg acaagaaaca
     2461 tgaacggcac cccatctttg gaaacatagt agatgaggtg gcatatcatg aaaagtaccc
     2521 aacgatttat cacctcagaa aaaagctagt tgactcaact gataaagcgg acctgaggtt
     2581 aatctacttg gctcttgccc atatgataaa gttccgtggg cactttctca ttgagggtga
     2641 tctaaatccg gacaactcgg atgtcgacaa actgttcatc cagttagtac aaacctataa
     2701 tcagttgttt gaagagaacc ctataaatgc aagtggcgtg gatgcgaagg ctattcttag
     2761 cgcccgcctc tctaaatccc gacggctaga aaacctgatc gcacaattac ccggagagaa
     2821 gaaaaatggg ttgttcggta accttatagc gctctcacta ggcctgacac caaattttaa
     2881 gtcgaacttc gacttagctg aagatgccaa attgcagctt agtaaggaca cgtacgatga
     2941 cgatctcgac aatctactgg cacaaattgg agatcagtat gcggacttat ttttggctgc
     3001 caaaaacctt agcgatgcaa tcctcctatc tgacatactg agagttaata ctgagattac
     3061 caaggcgccg ttatccgctt caatgatcaa aaggtacgat gaacatcacc aagacttgac
     3121 acttctcaag gccctagtcc gtcagcaact gcctgagaaa tataaggaaa tattctttga
     3181 tcagtcgaaa aacgggtacg caggttatat tgacggcgga gcgagtcaag aggaattcta
     3241 caagtttatc aaacccatat tagagaagat ggatgggacg gaagagttgc ttgtaaaact
     3301 caatcgcgaa gatctactgc gaaagcagcg gactttcgac aacggtagca ttccacatca
     3361 aatccactta ggcgaattgc atgctatact tagaaggcag gaggattttt atccgttcct
     3421 caaagacaat cgtgaaaaga ttgagaaaat cctaaccttt cgcatacctt actatgtggg
     3481 acccctggcc cgagggaact ctcggttcgc atggatgaca agaaagtccg aagaaacgat
     3541 tactccctgg aattttgagg aagttgtcga taaaggtgcg tcagctcaat cgttcatcga
     3601 gaggatgacc gcctttgaca agaatttacc gaacgaaaaa gtattgccta agcacagttt
     3661 actttacgag tatttcacag tgtacaatga actcacgaaa gttaagtatg tcactgaggg
     3721 catgcgtaaa cccgcctttc taagcggaga acagaagaaa gcaatagtag atctgttatt
     3781 caagaccaac cgcaaagtga cagttaagca attgaaagag gactacttta agaaaattga
     3841 atgcttcgat tctgtcgaga tctccggggt agaagatcga tttaatgcgt cacttggtac
     3901 gtatcatgac ctcctaaaga taattaaaga taaggacttc ctggataacg aagagaatga
     3961 agatatctta gaagatatag tgttgactct taccctcttt gaagatcggg aaatgattga
     4021 ggaaagacta aaaacatacg ctcacctgtt cgacgataag gttatgaaac agttaaagag
     4081 gcgtcgctat acgggctggg gagccttgtc gcggaaactt atcaacggga taagagacaa
     4141 gcaaagtggt aaaactattc tcgattttct aaagagcgac ggcttcgcca ataggaactt
     4201 tatggccctg atccatgatg actctttaac cttcaaagag gatatacaaa aggcacaggt
     4261 ttccggacaa ggggactcat tgcacgaaca tattgcgaat cttgctggtt cgccagccat
     4321 caaaaagggc atactccaga cagtcaaagt agtggatgag ctagttaagg tcatgggacg
     4381 tcacaaaccg gaaaacattg taatcgagat ggcacgcgaa aatcaaacga ctcagaaggg
     4441 gcaaaaaaac agtcgagagc ggatgaagag aatagaagag ggtattaaag aactgggcag
     4501 ccagatctta aaggagcatc ctgtggaaaa tacccaattg cagaacgaga aactttacct
     4561 ctattaccta caaaatggaa gggacatgta tgttgatcag gaactggaca taaaccgttt
     4621 atctgattac gacgtcgatc acattgtacc ccaatccttt ttgaaggacg attcaatcga
     4681 caataaagtg cttacacgct cggataagaa ccgagggaaa agtgacaatg ttccaagcga
     4741 ggaagtcgta aagaaaatga agaactattg gcggcagctc ctaaatgcga aactgataac
     4801 gcaaagaaag ttcgataact taactaaagc tgagaggggt ggcttgtctg aacttgacaa
     4861 ggccggattt attaaacgtc agctcgtgga aacccgcgcc atcacaaagc atgttgcgca
     4921 gatactagat tcccgaatga atacgaaata cgacgagaac gataagctga ttcgggaagt
     4981 caaagtaatc actttaaagt caaaattggt gtcggacttc agaaaggatt ttcaattcta
     5041 taaagttagg gagataaata actaccacca tgcgcacgac gcttatctta atgccgtcgt
     5101 agggaccgca ctcattaaga aatacccgaa gctagaaagt gagtttgtgt atggtgatta
     5161 caaagtttat gacgtccgta agatgatcgc gaaaagcgaa caggagatag gcaaggctac
     5221 agccaaatac ttcttttatt ctaacattat gaatttcttt aagacggaaa tcactctggc
     5281 aaacggagag atacgcaaac gacctttaat tgaaaccaat ggggagacag gtgaaatcgt
     5341 atgggataag ggccgggact tcgcgacggt gagaaaagtt ttgtccatgc cccaagtcaa
     5401 catagtaaag aaaactgagg tgcagaccgg agggttttca aaggaatcga ttcttccaaa
     5461 aaggaatagt gataagctca tcgctcgtaa aaaggactgg gacccgaaaa agtacggtgg
     5521 cttcgatagc cctacagttg cctattctgt cctagtagtg gcaaaagttg agaagggaaa
     5581 atccaagaaa ctgaagtcag tcaaagaatt attggggata acgattatgg agcgctcgtc
     5641 ttttgaaaag aaccccatcg acttccttga ggcgaaaggt tacaaggaag taaaaaagga
     5701 tctcataatt aaactaccaa agtatagtct gtttgagtta gaaaatggcc gaaaacggat
     5761 gttggctagc gccggagagc ttcaaaaggg gaacgaactc gcactaccgt ctaaatacgt
     5821 gaatttcctg tatttagcgt cccattacga gaagttgaaa ggttcacctg aagataacga
     5881 acagaagcaa ctttttgttg agcagcacaa acattatctc gacgaaatca tagagcaaat
     5941 ttcggaattc agtaagagag tcatcctagc tgatgccaat ctggacaaag tattaagcgc
     6001 atacaacaag cacagggata aacccatacg tgagcaggcg gaaaatatta tccatttgtt
     6061 tactcttacc aacctcggcg ctccagccgc attcaagtat tttgacacaa cgatagatcg
     6121 caaacgatac acttctacca aggaggtgct agacgcgaca ctgattcacc aatccatcac
     6181 gggattatat gaaactcgga tagatttgtc acagcttggg ggtgacggat cccccaagaa
     6241 gaagaggaaa gtctcgagcg actacaaaga ccatgacggt gattataaag atcatgacat
     6301 cgattacaag gatgacgatg acaaggctgc aggagctagc ggcagcggcg ccaccaactt
     6361 cagcctgctg aagcaggccg gcgacgtgga ggagaacccc ggccccatga ccgagtacaa
     6421 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     6481 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     6541 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     6601 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     6661 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     6721 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     6781 ggccaccgtc ggagtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     6841 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     6901 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     6961 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgaacgc gttaagtcga
     7021 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     7081 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     7141 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     7201 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     7261 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     7321 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     7381 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     7441 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     7501 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     7561 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc gtcgacttta
     7621 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     7681 ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct
     7741 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     7801 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     7861 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtacg
     7921 tatagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     7981 gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     8041 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     8101 atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg cccatcccgc
     8161 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     8221 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     8281 ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     8341 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     8401 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     8461 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     8521 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     8581 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     8641 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     8701 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     8761 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     8821 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     8881 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     8941 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     9001 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     9061 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     9121 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     9181 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     9241 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     9301 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     9361 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     9421 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     9481 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     9541 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     9601 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     9661 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     9721 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     9781 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     9841 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     9901 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     9961 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
    10021 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
    10081 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
    10141 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
    10201 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
    10261 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
    10321 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
    10381 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
    10441 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
    10501 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
    10561 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
    10621 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
    10681 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
    10741 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
    10801 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
    10861 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
    10921 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
    10981 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
    11041 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
    11101 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg
    11161 caagcttaat gtagtcttat gcaatact
//
