LOCUS       sequence-212226-       13357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for constitutive expression of BE3RA-P2A-GFP in 
            mammalian cells (codon optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13357)
  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 13357)
  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-BE3RA-P2A-GFP-PGK-Puro
FEATURES             Location/Qualifiers
     source          1..13357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..237
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             238..418
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    465..590
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1083..1316
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1501..1545
                     /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    1729..1846
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1915..2126
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2152..2838
                     /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(2887..2910,2911..2913,2914..6987)
                     /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             2887..6987
                     /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             7000..7248
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7261..7281
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7297..7353
                     /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(7354..7356,7357..7359,7360..8073)
                     /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(7660..7679)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8007..8028
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        8085..8584
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(8120..8143)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     8517..8536
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             8605..9204
                     /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(8605..8624)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     9101..9121
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    9220..9808
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9273..9293)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9691..9702)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9880..10113
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    10191..10312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10228..10247)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10282..10301
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10352..10487
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10507..10526)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10508..10526)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10536..10553)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10536..10552)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10545..10567)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10694..11149
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10781..10800)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10991..11012
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        11175..11279
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11280..11348,11349..12140)
                     /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(11498..11517)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12311..12899
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12800..12819
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13053..13070
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    13187..13208
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(13223..13228,13229..13246,13247..13253)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13261..13277
                     /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     13266..13288
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     13285..13301
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13285..13301
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     13320..13340
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        13322..13340
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt
       61 tagcaacatg ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg
      121 tggtacgatc gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc
      181 actgaattgc cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg
      241 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      301 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      361 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      421 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      481 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      541 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      601 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
      661 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
      721 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
      781 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
      841 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
      901 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
      961 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1021 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1081 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1141 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1201 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1261 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1321 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1381 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1441 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1501 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1561 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1621 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     1681 ttatcgtttc agacccacct cccaaccccg aggggaccca gaccacaatt ttaaaagaaa
     1741 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat
     1801 acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag
     1861 ggacagcaga gatccacttt ggcgccggct cgagtggctc cggtgcccgt cagtgggcag
     1921 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg
     1981 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     2041 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     2101 gcaacgggtt tgccgccaga acacaggtgt cgtgacgcgg gatccgccac catgagctca
     2161 gagactggcc cagtggctgt ggaccccaca ttgagacggc ggatcgagcc ccatgagttt
     2221 gaggtattct tcgatccgag agagctccgc aaggagacct gcctgcttta cgaaattaat
     2281 tgggggggcc ggcactccat ttggcgacat acatcacaga acactaacaa gcacgtcgaa
     2341 gtcaacttca tcgagaagtt cacgacagaa agatatttct gtccgaacac aaggtgcagc
     2401 attacctggt ttctcagctg gagcccatgc ggcgaatgta gtagggccat cactgaattc
     2461 ctgtcaaggt atccccacgt cactctgttt atttacatcg caaggctgta ccaccacgct
     2521 gacccccgca atcgacaagg cctgcgggat ttgatctctt caggtgtgac tatccaaatt
     2581 atgactgagc aggagtcagg atactgctgg agaaactttg tgaattatag cccgagtaat
     2641 gaagcccact ggcctaggta tccccatctg tgggtacgac tgtacgttct tgaactgtac
     2701 tgcatcatac tgggcctgcc tccttgtctc aacattctga gaaggaagca gccacagctg
     2761 acattcttta ccatcgctct tcagtcttgt cattaccagc gactgccccc acacattctc
     2821 tgggccaccg ggttgaaaag cggcagcgag actcccggga cctcagagtc cgccacaccc
     2881 gaaagtgaca agaagtacag catcggcctg gccatcggca ccaactctgt gggctgggcc
     2941 gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac
     3001 cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc
     3061 gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc
     3121 tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac
     3181 agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc
     3241 ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga
     3301 aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc
     3361 cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc
     3421 gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac
     3481 cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc
     3541 agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcggc
     3601 aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc
     3661 gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg
     3721 gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc
     3781 atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc
     3841 tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg
     3901 cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac
     3961 gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc
     4021 ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg
     4081 cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg
     4141 cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag
     4201 atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac
     4261 agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag
     4321 gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat
     4381 aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc
     4441 gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc
     4501 ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg
     4561 accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa
     4621 atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa
     4681 attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc
     4741 gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat
     4801 gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg
     4861 ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc
     4921 ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac
     4981 gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc
     5041 ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag
     5101 acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc
     5161 gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag
     5221 agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac
     5281 ccagtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg
     5341 cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac
     5401 catatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga
     5461 agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg
     5521 aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat
     5581 ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga
     5641 cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg
     5701 aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag
     5761 tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac
     5821 aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa
     5881 aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg
     5941 aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac
     6001 agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag
     6061 cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat
     6121 tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag
     6181 gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg
     6241 atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg
     6301 gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt
     6361 gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc
     6421 gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct
     6481 aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa
     6541 ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc
     6601 agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg
     6661 gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga
     6721 gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat
     6781 aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga
     6841 gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc
     6901 aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg
     6961 atcgacctgt ctcagctggg aggcgattca ggcggatcta ctaatctgtc agatattatt
     7021 gaaaaggaga ccggtaagca actggttatc caggaatcca tcctcatgct cccagaggag
     7081 gtggaagaag tcattgggaa caagccggaa agcgatatac tcgtgcacac cgcctacgac
     7141 gagagcaccg acgagaatgt catgcttctg actagcgacg cccctgaata caagccttgg
     7201 gctctggtca tacaggatag caacggtgag aacaagatta agatgctctc tggtggttct
     7261 cccaagaaga agaggaaagt cgctagcggc agcggcgcca ccaacttcag cctgctgaag
     7321 caggccggcg acgtggagga gaaccccggc cccatggtga gcaagggcga ggagctgttc
     7381 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     7441 gtgtctggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     7501 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     7561 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     7621 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     7681 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     7741 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     7801 aacgtctata tcatggccga caagcagaag aacggcatca aggcgaactt caagatccgc
     7861 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     7921 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     7981 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     8041 atcactctcg gcatggacga gctgtacaag taaacgcgtc taccgggtag gggaggcgct
     8101 tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac
     8161 aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt
     8221 ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc
     8281 cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt
     8341 gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata
     8401 gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg
     8461 gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt
     8521 ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct
     8581 ttcgacctgc agcccaagct taccatgacc gagtacaagc ccacggtgcg cctcgccacc
     8641 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     8701 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     8761 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     8821 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     8881 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     8941 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     9001 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     9061 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     9121 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     9181 atgacccgca agcccggtgc ctgattctag aaagtcgaca atcaacctct ggattacaaa
     9241 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     9301 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     9361 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     9421 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     9481 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     9541 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     9601 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     9661 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     9721 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     9781 cggatctccc tttgggccgc ctccccgcgt cgactttaag accaatgact tacaaggcag
     9841 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     9901 aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     9961 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
    10021 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
    10081 gaccctttta gtcagtgtgg aaaatctcta gcagtacgta tagtagttca tgtcatctta
    10141 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta
    10201 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
    10261 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
    10321 ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
    10381 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
    10441 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc
    10501 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
    10561 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
    10621 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
    10681 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
    10741 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
    10801 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
    10861 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
    10921 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
    10981 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
    11041 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
    11101 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
    11161 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
    11221 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
    11281 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
    11341 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
    11401 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
    11461 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
    11521 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
    11581 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
    11641 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
    11701 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
    11761 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
    11821 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
    11881 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
    11941 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
    12001 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
    12061 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
    12121 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
    12181 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
    12241 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
    12301 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
    12361 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
    12421 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
    12481 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
    12541 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
    12601 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
    12661 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
    12721 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
    12781 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
    12841 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
    12901 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
    12961 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
    13021 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
    13081 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
    13141 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
    13201 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
    13261 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
    13321 caattaaccc tcactaaagg gaacaaaagc tggagct
//
