LOCUS       sequence-211476-       13477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for constitutive expression of FNLS-P2A-GFP in 
            mammalian cells (codon optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13477)
  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 13477)
  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-FNLS-P2A-GFP-PGK-Puro
FEATURES             Location/Qualifiers
     source          1..13477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1719..1836
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1905..2116
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2136..2145
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2145..2210
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2217..2237
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2262..2948
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             join(2997..3020,3021..3023,3024..7097)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             2997..7097
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7110..7358
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7371..7391
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7407..7463
                     /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(7464..7466,7467..7469,7470..8183)
                     /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(7770..7789)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8117..8138
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        8195..8694
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8230..8253)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     8627..8646
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             8715..9314
                     /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(8715..8734)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9211..9231
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9330..9918
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9383..9403)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9801..9812)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9990..10223
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    10301..10422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10338..10357)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10392..10411
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10462..10597
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10617..10636)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10618..10636)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10646..10663)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10646..10662)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10655..10677)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10804..11259
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10891..10910)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11101..11122
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        11285..11389
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11390..11458,11459..12250)
                     /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(11608..11627)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12421..13009
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12910..12929
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13163..13180
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    13297..13318
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(13333..13338,13339..13356,13357..13363)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13371..13387
                     /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     13376..13398
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     13395..13411
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13395..13411
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     13430..13450
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        13432..13450
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattaaat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agacccacct cccaaccccg aggggaccca gaccacaatt ttaaaagaaa aggggggatt
     1741 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     1801 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga
     1861 gatccacttt ggcgccggct cgagtggctc cggtgcccgt cagtgggcag agcgcacatc
     1921 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag
     1981 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     2041 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     2101 tgccgccaga acacaggtgt cgtgacgcgg gatccgccac catggactat aaggaccacg
     2161 acggagacta caaggatcat gatattgatt acaaagacga tgacgataag atggccccaa
     2221 agaagaagcg gaaggtcggt atccacggag tcccagcagc catgagctca gagactggcc
     2281 cagtggctgt ggaccccaca ttgagacggc ggatcgagcc ccatgagttt gaggtattct
     2341 tcgatccgag agagctccgc aaggagacct gcctgcttta cgaaattaat tgggggggcc
     2401 ggcactccat ttggcgacat acatcacaga acactaacaa gcacgtcgaa gtcaacttca
     2461 tcgagaagtt cacgacagaa agatatttct gtccgaacac aaggtgcagc attacctggt
     2521 ttctcagctg gagcccatgc ggcgaatgta gtagggccat cactgaattc ctgtcaaggt
     2581 atccccacgt cactctgttt atttacatcg caaggctgta ccaccacgct gacccccgca
     2641 atcgacaagg cctgcgggat ttgatctctt caggtgtgac tatccaaatt atgactgagc
     2701 aggagtcagg atactgctgg agaaactttg tgaattatag cccgagtaat gaagcccact
     2761 ggcctaggta tccccatctg tgggtacgac tgtacgttct tgaactgtac tgcatcatac
     2821 tgggcctgcc tccttgtctc aacattctga gaaggaagca gccacagctg acattcttta
     2881 ccatcgctct tcagtcttgt cattaccagc gactgccccc acacattctc tgggccaccg
     2941 ggttgaaaag cggcagcgag actcccggga cctcagagtc cgccacaccc gaaagtgaca
     3001 agaagtacag catcggcctg gccatcggca ccaactctgt gggctgggcc gtgatcaccg
     3061 acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca
     3121 tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc
     3181 ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc
     3241 aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag
     3301 agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg
     3361 tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg
     3421 tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca
     3481 agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca
     3541 agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg
     3601 ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg
     3661 aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcggc aacctgattg
     3721 ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca
     3781 aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg
     3841 gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga
     3901 gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca
     3961 agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc
     4021 tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca
     4081 ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga
     4141 tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc
     4201 ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg cacgccattc
     4261 tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga
     4321 tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg
     4381 cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg
     4441 acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc
     4501 ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg
     4561 agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg
     4621 agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc
     4681 agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg
     4741 tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg
     4801 acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc
     4861 tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt
     4921 tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga
     4981 gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc
     5041 tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga
     5101 cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc
     5161 acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg
     5221 tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa
     5281 tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc
     5341 ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac ccagtggaaa
     5401 acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt
     5461 acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc
     5521 ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga
     5581 accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact
     5641 ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg
     5701 ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg
     5761 aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt
     5821 acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg
     5881 tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc
     5941 acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta
     6001 agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg
     6061 ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca
     6121 tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga
     6181 tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg
     6241 tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag
     6301 gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa
     6361 agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg
     6421 tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc
     6481 tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg
     6541 aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc
     6601 tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg
     6661 gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg
     6721 agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca
     6781 agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg
     6841 ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca
     6901 gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg
     6961 ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc
     7021 tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt
     7081 ctcagctggg aggcgattca ggcggatcta ctaatctgtc agatattatt gaaaaggaga
     7141 ccggtaagca actggttatc caggaatcca tcctcatgct cccagaggag gtggaagaag
     7201 tcattgggaa caagccggaa agcgatatac tcgtgcacac cgcctacgac gagagcaccg
     7261 acgagaatgt catgcttctg actagcgacg cccctgaata caagccttgg gctctggtca
     7321 tacaggatag caacggtgag aacaagatta agatgctctc tggtggttct cccaagaaga
     7381 agaggaaagt catgcatggc agcggcgcca ccaacttcag cctgctgaag caggccggcg
     7441 acgtggagga gaaccccggc cccatggtga gcaagggcga ggagctgttc accggggtgg
     7501 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtctggcg
     7561 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     7621 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     7681 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     7741 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     7801 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     7861 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     7921 tcatggccga caagcagaag aacggcatca aggcgaactt caagatccgc cacaacatcg
     7981 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     8041 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     8101 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     8161 gcatggacga gctgtacaag taagctagcc taccgggtag gggaggcgct tttcccaagg
     8221 cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc
     8281 tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc
     8341 cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc
     8401 gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac
     8461 agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc
     8521 tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc
     8581 gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt ctgcacgctt
     8641 caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct ttcgacctgc
     8701 agcccaagct taccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     8761 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     8821 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     8881 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     8941 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     9001 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     9061 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     9121 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     9181 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     9241 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     9301 agcccggtgc ctgattctag aaagtcgaca atcaacctct ggattacaaa atttgtgaaa
     9361 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     9421 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     9481 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     9541 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     9601 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     9661 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     9721 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     9781 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     9841 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     9901 tttgggccgc ctccccgcgt cgactttaag accaatgact tacaaggcag ctgtagatct
     9961 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
    10021 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
    10081 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
    10141 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
    10201 gtcagtgtgg aaaatctcta gcagtacgta tagtagttca tgtcatctta ttattcagta
    10261 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
    10321 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
    10381 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
    10441 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
    10501 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
    10561 ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct
    10621 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
    10681 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
    10741 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
    10801 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
    10861 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
    10921 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
    10981 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
    11041 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
    11101 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
    11161 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
    11221 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
    11281 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
    11341 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
    11401 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
    11461 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
    11521 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
    11581 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
    11641 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
    11701 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
    11761 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
    11821 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
    11881 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
    11941 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
    12001 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
    12061 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
    12121 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
    12181 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
    12241 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
    12301 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
    12361 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
    12421 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
    12481 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
    12541 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
    12601 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
    12661 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
    12721 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
    12781 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
    12841 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
    12901 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
    12961 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
    13021 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
    13081 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
    13141 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
    13201 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
    13261 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
    13321 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
    13381 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
    13441 tcactaaagg gaacaaaagc tggagctgca agcttaa
//
