LOCUS       sequence-208409-       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        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..3633,3634..3636,3637..4125,4126..4128,4129..4227,
                     4228..4230,4231..4920,4921..4923,4924..6249)
                     /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             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"
     promoter        6364..6863
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6399..6422)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6796..6815
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6884..7483
                     /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(6884..6903)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7380..7400
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7499..8087
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7552..7572)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7970..7981)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8159..8392
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8470..8591
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8507..8526)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8561..8580
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8631..8766
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8786..8805)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8787..8805)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8815..8832)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8815..8831)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8824..8846)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8973..9428
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9060..9079)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9270..9291
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9454..9558
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9559..9627,9628..10419)
                     /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(9777..9796)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10590..11178
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11079..11098
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11332..11349
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11466..11487
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11502..11507,11508..11525,11526..11532)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11540..11556
                     /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     11545..11567
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11564..11580
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11564..11580
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11599..11619
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11601..11619
                     /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 gattactccc tggaattttg aggaagttgt cgataaaggt
     3601 gcgtcagctc aatcgttcat cgagaggatg accgcctttg 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 ggggagcctt gtcgcggaaa
     4141 cttatcaacg ggataagaga caagcaaagt ggtaaaacta ttctcgattt tctaaagagc
     4201 gacggcttcg ccaataggaa ctttatggcc 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 gccatcacaa agcatgttgc gcagatacta 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 tggcttcgat 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 agcgccggag 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 tcgcaaacga tacacttcta ccaaggaggt gctagacgcg
     6181 acactgattc accaatccat cacgggatta tatgaaactc ggatagattt gtcacagctt
     6241 gggggtgacg gatcccccaa gaagaagagg aaagtctcga gcgactacaa agaccatgac
     6301 ggtgattata aagatcatga catcgattac aaggatgacg atgacaagta agatgcatct
     6361 accgggtagg ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc
     6421 tgggcacttg gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta
     6481 ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag
     6541 tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag
     6601 cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc
     6661 tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa
     6721 ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt
     6781 cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct
     6841 cttcctcatc tccgggcctt tcgacctgca gcccaagctt accatgaccg agtacaagcc
     6901 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     6961 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     7021 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     7081 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     7141 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     7201 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     7261 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     7321 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     7381 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     7441 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgattctaga aagtcgacaa
     7501 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     7561 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     7621 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     7681 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     7741 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     7801 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     7861 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     7921 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     7981 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     8041 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcgtc gactttaaga
     8101 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     8161 gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac tgggtctctc
     8221 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     8281 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     8341 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtacgtat
     8401 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     8461 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     8521 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     8581 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc
     8641 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     8701 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     8761 gaggcctagg gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc
     8821 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     8881 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     8941 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     9001 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     9061 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     9121 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     9181 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     9241 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     9301 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     9361 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     9421 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     9481 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     9541 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     9601 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     9661 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     9721 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     9781 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     9841 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     9901 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     9961 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
    10021 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
    10081 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
    10141 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
    10201 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
    10261 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
    10321 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
    10381 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
    10441 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
    10501 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
    10561 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
    10621 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
    10681 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
    10741 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
    10801 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
    10861 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
    10921 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
    10981 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
    11041 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
    11101 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
    11161 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
    11221 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
    11281 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
    11341 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
    11401 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
    11461 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
    11521 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
    11581 catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa
    11641 gc
//
