LOCUS       sequence-211472-       12448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for constitutive expression of Cas9-HF1RA-P2A-GFP 
            in mammalian cells (codon optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12448)
  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 12448)
  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-HF1RA-P2A-GFP-PGK-Puro
FEATURES             Location/Qualifiers
     source          1..12448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             222..402
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    449..574
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1067..1300
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1485..1529
                     /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    1713..1830
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1899..2110
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2130..2139
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2139..2162
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2169..2189
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2214..6314
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGALSRKLINGIRDKQSGKTILDFLKSDGFANRNFMA
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRAITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6315..6362
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6372..6428
                     /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(6429..6431,6432..6434,6435..7148)
                     /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(6735..6754)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7082..7103
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        7160..7659
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7195..7218)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7592..7611
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             7680..8279
                     /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(7680..7699)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8176..8196
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8295..8883
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8348..8368)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8766..8777)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8955..9188
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9266..9387
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9303..9322)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9357..9376
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9427..9562
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9582..9601)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9583..9601)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9611..9628)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9611..9627)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9620..9642)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9769..10224
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9856..9875)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10066..10087
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10250..10354
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10355..10423,10424..11215)
                     /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(10573..10592)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11386..11974
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11875..11894
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12128..12145
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12262..12283
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12298..12303,12304..12321,12322..12328)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12336..12352
                     /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     12341..12363
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12360..12376
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12360..12376
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12395..12415
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        12397..12415
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        join(12443..12448,1..221)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa catgccttac
       61 aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac gatcgtgcct
      121 tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa ttgccgcatt
      181 gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc tggttagacc
      241 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
      301 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
      361 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
      421 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
      481 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
      541 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
      601 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
      661 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
      721 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
      781 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
      841 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
      901 cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
      961 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     1021 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     1081 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     1141 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     1201 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     1261 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     1321 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     1381 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     1441 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     1501 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     1561 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     1621 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     1681 acctcccaac cccgagggga cccagaccac aattttaaaa gaaaaggggg gattgggggg
     1741 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
     1801 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
     1861 ctttggcgcc ggctcgagtg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac
     1921 agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg
     1981 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg
     2041 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc
     2101 cagaacacag gtgtcgtgac gcgggatccg ccaccatgga ttacaaagac gatgacgata
     2161 agatggcccc aaagaagaag cggaaggtcg gtatccacgg agtcccagca gccgacaaga
     2221 agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg atcaccgacg
     2281 agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca
     2341 agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc
     2401 tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag
     2461 agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt
     2521 ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg
     2581 acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg
     2641 acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt
     2701 tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc
     2761 tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca
     2821 gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa
     2881 atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac ctgattgccc
     2941 tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac
     3001 tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg
     3061 accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg
     3121 acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga
     3181 gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc
     3241 ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg
     3301 acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg
     3361 acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga
     3421 ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc
     3481 ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc
     3541 tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct
     3601 ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca
     3661 agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca
     3721 acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc
     3781 tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc
     3841 agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc
     3901 tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg
     3961 aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca
     4021 aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga
     4081 cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg
     4141 acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc gccctgagcc
     4201 ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga
     4261 agtccgacgg cttcgccaac agaaacttca tggccctgat ccacgacgac agcctgacct
     4321 ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca
     4381 ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg
     4441 tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg
     4501 ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga
     4561 tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca
     4621 cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg
     4681 tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc
     4741 agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc
     4801 ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc
     4861 ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg
     4921 agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa
     4981 cccgggccat cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg
     5041 acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt
     5101 ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg
     5161 cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc
     5221 tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca
     5281 agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga
     5341 actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg
     5401 agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc
     5461 ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg
     5521 gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga
     5581 aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc
     5641 tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc
     5701 tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag
     5761 ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt
     5821 tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa
     5881 acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga
     5941 agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc
     6001 actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg
     6061 acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag
     6121 agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct
     6181 tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg
     6241 acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc
     6301 agctgggagg cgacaagcgt cctgctgcta ctaagaaagc tggtcaagct aagaaaaaga
     6361 aaggcagcgg cgccaccaac ttcagcctgc tgaagcaggc cggcgacgtg gaggagaacc
     6421 ccggccccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     6481 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc tggcgagggc gagggcgatg
     6541 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     6601 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     6661 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     6721 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     6781 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     6841 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     6901 agaagaacgg catcaaggcg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     6961 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     7021 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     7081 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     7141 acaagtaagc tagcctaccg ggtaggggag gcgcttttcc caaggcagtc tggagcatgc
     7201 gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc tcgcacacat
     7261 tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt cgcgccacct
     7321 tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg
     7381 tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac cgctgagcaa
     7441 tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt cgctttctgg
     7501 gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct caggggcggg
     7561 gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct
     7621 gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcagccc aagcttacca
     7681 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     7741 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     7801 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     7861 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     7921 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     7981 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     8041 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     8101 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     8161 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     8221 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgat
     8281 tctagaaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     8341 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     8401 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     8461 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     8521 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     8581 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     8641 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     8701 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     8761 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     8821 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     8881 cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     8941 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc
     9001 ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     9061 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     9121 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     9181 ctctagcagt acgtatagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     9241 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
     9301 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     9361 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
     9421 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     9481 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     9541 tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt
     9601 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     9661 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     9721 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     9781 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     9841 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     9901 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     9961 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
    10021 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
    10081 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
    10141 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
    10201 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
    10261 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
    10321 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
    10381 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
    10441 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
    10501 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
    10561 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
    10621 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
    10681 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
    10741 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
    10801 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
    10861 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
    10921 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
    10981 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
    11041 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
    11101 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
    11161 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
    11221 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
    11281 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
    11341 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
    11401 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
    11461 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
    11521 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
    11581 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
    11641 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
    11701 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
    11761 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
    11821 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
    11881 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
    11941 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
    12001 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
    12061 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
    12121 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
    12181 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
    12241 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
    12301 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
    12361 aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca
    12421 aaagctggag ctgcaagctt aatgtagt
//
