LOCUS       sequence-212266-       12508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus for constitutive expression of BE4RA in mammalian cells 
            (codon optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12508)
  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 12508)
  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-BE4RA-P2A-Puro
FEATURES             Location/Qualifiers
     source          1..12508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..231
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             232..412
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    459..584
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1077..1310
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1495..1539
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1723..1840
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1909..2120
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2140..2149
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2149..2214
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2221..2241
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2266..2952
                     /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(3001..3024,3025..3027,3028..7101)
                     /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             3001..7101
                     /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             7114..7362
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7375..7395
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7396..7644
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7657..7677
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7693..7749
                     /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             7750..8349
                     /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(7750..7769)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8246..8266
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8365..8953
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8418..8438)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8836..8847)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9025..9258
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9336..9457
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9373..9392)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9427..9446
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9497..9632
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9652..9671)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9653..9671)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9681..9698)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9681..9697)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9690..9712)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9839..10294
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9926..9945)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10136..10157
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10320..10424
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10425..10493,10494..11285)
                     /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(10643..10662)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11456..12044
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11945..11964
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12198..12215
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12332..12353
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12368..12373,12374..12391,12392..12398)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12406..12422
                     /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     12411..12433
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12430..12446
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12430..12446
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12465..12485
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        12467..12485
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
       61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
      121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
      181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
      241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
      301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
      361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
      421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
      481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
      541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
      601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
      661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
      721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
      781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
      841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
      901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
      961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     1681 tttcagaccc acctcccaac cccgagggga cccagaccac aattttaaaa gaaaaggggg
     1741 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     1801 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     1861 cagagatcca ctttggcgcc ggctcgagtg gctccggtgc ccgtcagtgg gcagagcgca
     1921 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga
     1981 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
     2041 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
     2101 ggtttgccgc cagaacacag gtgtcgtgac gcgggatccg ccaccatgga ctataaggac
     2161 cacgacggag actacaagga tcatgatatt gattacaaag acgatgacga taagatggcc
     2221 ccaaagaaga agcggaaggt cggtatccac ggagtcccag cagccatgag ctcagagact
     2281 ggcccagtgg ctgtggaccc cacattgaga cggcggatcg agccccatga gtttgaggta
     2341 ttcttcgatc cgagagagct ccgcaaggag acctgcctgc tttacgaaat taattggggg
     2401 ggccggcact ccatttggcg acatacatca cagaacacta acaagcacgt cgaagtcaac
     2461 ttcatcgaga agttcacgac agaaagatat ttctgtccga acacaaggtg cagcattacc
     2521 tggtttctca gctggagccc atgcggcgaa tgtagtaggg ccatcactga attcctgtca
     2581 aggtatcccc acgtcactct gtttatttac atcgcaaggc tgtaccacca cgctgacccc
     2641 cgcaatcgac aaggcctgcg ggatttgatc tcttcaggtg tgactatcca aattatgact
     2701 gagcaggagt caggatactg ctggagaaac tttgtgaatt atagcccgag taatgaagcc
     2761 cactggccta ggtatcccca tctgtgggta cgactgtacg ttcttgaact gtactgcatc
     2821 atactgggcc tgcctccttg tctcaacatt ctgagaagga agcagccaca gctgacattc
     2881 tttaccatcg ctcttcagtc ttgtcattac cagcgactgc ccccacacat tctctgggcc
     2941 accgggttga aaagcggcag cgagactccc gggacctcag agtccgccac acccgaaagt
     3001 gacaagaagt acagcatcgg cctggccatc ggcaccaact ctgtgggctg ggccgtgatc
     3061 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
     3121 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
     3181 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
     3241 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
     3301 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
     3361 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
     3421 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
     3481 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
     3541 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
     3601 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
     3661 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggcaacctg
     3721 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
     3781 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
     3841 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
     3901 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
     3961 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
     4021 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
     4081 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
     4141 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
     4201 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
     4261 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
     4321 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
     4381 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
     4441 gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac
     4501 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat
     4561 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
     4621 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
     4681 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
     4741 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
     4801 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
     4861 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
     4921 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg
     4981 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
     5041 ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc
     5101 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
     5161 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
     5221 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
     5281 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg
     5341 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
     5401 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
     5461 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc
     5521 gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac
     5581 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
     5641 tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc
     5701 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
     5761 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
     5821 aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag
     5881 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
     5941 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
     6001 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
     6061 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac
     6121 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct
     6181 ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc
     6241 accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag
     6301 acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc
     6361 agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat
     6421 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa
     6481 gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt
     6541 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac
     6601 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag
     6661 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac
     6721 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag
     6781 cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc
     6841 ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc
     6901 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct
     6961 gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag
     7021 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac
     7081 ctgtctcagc tgggaggcga ctctggtggt tctactaatc tgtcagatat tattgaaaag
     7141 gagaccggta agcaactggt tatccaggaa tccatcctca tgctcccaga ggaggtggaa
     7201 gaagtcattg ggaacaagcc ggaaagcgat atactcgtgc acaccgccta cgacgagagc
     7261 accgacgaga atgtcatgct tctgactagc gacgcccctg aatacaagcc ttgggctctg
     7321 gtcatacagg atagcaacgg tgagaacaag attaagatgc tctctggtgg ttctcccaag
     7381 aagaagagga aagtcacaaa tctctctgac atcatagaga aggagacagg gaaacaactc
     7441 gtaatacaag agtccattct tatgctccct gaggaggtgg aagaagttat cggcaacaaa
     7501 ccagagagtg acattctggt ccataccgcc tacgatgaaa gcacagacga gaacgttatg
     7561 ttgctcactt ctgacgctcc agaatacaaa ccttgggcac tcgtcattca ggacagcaac
     7621 ggcgagaaca agatcaaaat gcttagcggg ggcagcccca aaaaaaagag gaaggtcgct
     7681 agcggcagcg gcgccaccaa cttcagcctg ctgaagcagg ccggcgacgt ggaggagaac
     7741 cccggcccca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     7801 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     7861 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     7921 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg cggccgtggc ggtctggacc
     7981 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     8041 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     8101 cccaaggagc ccgcgtggtt cctggccacc gtcggagtct cgcccgacca ccagggcaag
     8161 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     8221 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     8281 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     8341 ggtgcctgaa cgcgttaagt cgacaatcaa cctctggatt acaaaatttg tgaaagattg
     8401 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     8461 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     8521 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     8581 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     8641 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     8701 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     8761 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     8821 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     8881 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     8941 gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     9001 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     9061 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     9121 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     9181 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     9241 tgtggaaaat ctctagcagt acgtatagta gttcatgtca tcttattatt cagtatttat
     9301 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     9361 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     9421 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     9481 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     9541 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     9601 ccagaagtag tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt
     9661 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     9721 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     9781 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     9841 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     9901 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     9961 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
    10021 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
    10081 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
    10141 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
    10201 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
    10261 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
    10321 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
    10381 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
    10441 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
    10501 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
    10561 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
    10621 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
    10681 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
    10741 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
    10801 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
    10861 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
    10921 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
    10981 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
    11041 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
    11101 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
    11161 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
    11221 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
    11281 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
    11341 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
    11401 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
    11461 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
    11521 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
    11581 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
    11641 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
    11701 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
    11761 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
    11821 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
    11881 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
    11941 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
    12001 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
    12061 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
    12121 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
    12181 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
    12241 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
    12301 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
    12361 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
    12421 aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact
    12481 aaagggaaca aaagctggag ctgcaagc
//
