LOCUS       sequence-211469-       12448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for constitutive expression of Cas9-VQRRA-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-VQRRA-P2A-GFP-PGK-Puro
FEATURES             Location/Qualifiers
     source          1..12448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             221..401
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    448..573
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1066..1299
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1484..1528
                     /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    1712..1829
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1898..2109
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2129..2138
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2138..2161
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2168..2188
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2213..5611,5612..5614,5615..6211,6212..6214,6215..6217,
                     6218..6220,6221..6313)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             2213..6313
                     /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
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6314..6361
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6371..6427
                     /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(6428..6430,6431..6433,6434..7147)
                     /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(6734..6753)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7081..7102
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        7159..7658
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7194..7217)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     7591..7610
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             7679..8278
                     /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(7679..7698)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8175..8195
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8294..8882
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8347..8367)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8765..8776)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8954..9187
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9265..9386
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9302..9321)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9356..9375
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9426..9561
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9581..9600)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9582..9600)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9610..9627)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9610..9626)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9619..9641)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9768..10223
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9855..9874)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10065..10086
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10249..10353
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10354..10422,10423..11214)
                     /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(10572..10591)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11385..11973
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11874..11893
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12127..12144
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12261..12282
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12297..12302,12303..12320,12321..12327)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12335..12351
                     /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     12340..12362
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12359..12375
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12359..12375
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12394..12414
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        12396..12414
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        join(12442..12448,1..220)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca
       61 aggagagaaa aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt
      121 attaggaagg caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg
      181 cagagatatt gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca
      241 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
      301 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
      361 atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac
      421 ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg
      481 cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga
      541 ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg
      601 cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata
      661 gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca
      721 gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa
      781 cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata
      841 aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc
      901 gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag
      961 aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa
     1021 ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct
     1081 tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca
     1141 ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga
     1201 ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat
     1261 cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg
     1321 aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga
     1381 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag
     1441 cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt
     1501 attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg
     1561 gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc
     1621 tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca
     1681 cctcccaacc ccgaggggac ccagaccaca attttaaaag aaaagggggg attggggggt
     1741 acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac
     1801 aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccac
     1861 tttggcgccg gctcgagtgg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca
     1921 gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag aaggtggcgc
     1981 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga
     2041 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc
     2101 agaacacagg tgtcgtgacg cgggatccgc caccatggat tacaaagacg atgacgataa
     2161 gatggcccca aagaagaagc ggaaggtcgg tatccacgga gtcccagcag ccgacaagaa
     2221 gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga
     2281 gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa
     2341 gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct
     2401 gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga
     2461 gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc
     2521 cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga
     2581 cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga
     2641 cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt
     2701 ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct
     2761 gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag
     2821 cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa
     2881 tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct
     2941 gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact
     3001 gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga
     3061 ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga
     3121 catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag
     3181 atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc
     3241 tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga
     3301 cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga
     3361 cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac
     3421 cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg
     3481 gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct
     3541 gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg
     3601 gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa
     3661 gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa
     3721 cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct
     3781 gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca
     3841 gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct
     3901 gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga
     3961 agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa
     4021 ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac
     4081 actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga
     4141 cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg
     4201 gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa
     4261 gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt
     4321 taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat
     4381 tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt
     4441 ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc
     4501 cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat
     4561 cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac
     4621 ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt
     4681 ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca
     4741 gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg
     4801 gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg
     4861 gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga
     4921 gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac
     4981 ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga
     5041 cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc
     5101 cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc
     5161 ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct
     5221 ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa
     5281 gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa
     5341 ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga
     5401 gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg
     5461 gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg
     5521 cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa
     5581 ggactgggac cctaagaagt acggcggctt cgtcagcccc accgtggcct attctgtgct
     5641 ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct
     5701 ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc
     5761 caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt
     5821 cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa
     5881 cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa
     5941 gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca
     6001 ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga
     6061 cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga
     6121 gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt
     6181 caagtacttt gacaccacca tcgaccggaa gcagtacagg agcaccaaag aggtgctgga
     6241 cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca
     6301 gctgggaggc gacaagcgtc ctgctgctac taagaaagct ggtcaagcta agaaaaagaa
     6361 aggcagcggc gccaccaact tcagcctgct gaagcaggcc ggcgacgtgg aggagaaccc
     6421 cggccccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     6481 gctggacggc gacgtaaacg gccacaagtt cagcgtgtct ggcgagggcg agggcgatgc
     6541 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     6601 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     6661 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     6721 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     6781 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     6841 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     6901 gaagaacggc atcaaggcga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     6961 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     7021 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     7081 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     7141 caagtaagct agcctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg
     7201 ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt
     7261 ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt
     7321 ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt
     7381 gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat
     7441 ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg
     7501 ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg
     7561 cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg
     7621 ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcagccca agcttaccat
     7681 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     7741 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     7801 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     7861 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     7921 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     7981 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     8041 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     8101 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     8161 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     8221 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgatt
     8281 ctagaaagtc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     8341 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     8401 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     8461 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     8521 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     8581 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     8641 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     8701 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     8761 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     8821 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     8881 gcgtcgactt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     8941 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct
     9001 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     9061 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     9121 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     9181 tctagcagta cgtatagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     9241 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
     9301 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     9361 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc
     9421 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     9481 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     9541 gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta
     9601 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     9661 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     9721 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     9781 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     9841 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     9901 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     9961 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
    10021 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
    10081 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
    10141 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
    10201 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
    10261 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
    10321 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
    10381 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
    10441 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
    10501 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
    10561 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
    10621 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
    10681 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
    10741 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
    10801 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
    10861 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
    10921 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
    10981 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
    11041 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
    11101 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
    11161 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
    11221 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
    11281 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
    11341 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
    11401 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
    11461 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
    11521 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
    11581 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
    11641 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
    11701 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
    11761 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
    11821 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
    11881 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
    11941 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
    12001 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
    12061 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
    12121 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
    12181 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
    12241 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
    12301 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
    12361 ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa
    12421 aagctggagc tgcaagctta atgtagtc
//
