LOCUS       sequence-260843-       12277 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for constitutive expression of xFNLS in mammalian 
            cells (codon optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12277)
  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 12277)
  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-xF2X-P2A-Puro
FEATURES             Location/Qualifiers
     source          1..12277
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    126..243
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        312..523
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      543..552
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             552..617
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             624..644
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             669..1355
                     /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             1374..1394
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1404..1424
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1455..5555
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 mutant with broad PAM
                     compatibility (Hu et al., 2018)"
                     /label=xCas9-3.7
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDTKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKLYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGIIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEKVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGDQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFIQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1455..5555
                     /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
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDTKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKLYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGIIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEKVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGDQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFIQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5568..5816
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5829..5849
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5865..5921
                     /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             5922..6521
                     /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(5922..5941)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6418..6438
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6537..7125
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6590..6610)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7008..7019)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7197..7430
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7508..7629
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7545..7564)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7599..7618
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7669..7804
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7824..7843)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7825..7843)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7853..7870)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7853..7869)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7862..7884)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8011..8466
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8098..8117)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8308..8329
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8492..8596
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8597..8665,8666..9457)
                     /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(8815..8834)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9628..10216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10117..10136
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10370..10387
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10504..10525
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10540..10545,10546..10563,10564..10570)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10578..10594
                     /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     10583..10605
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10602..10618
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10602..10618
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10637..10657
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10639..10657
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        10685..10911
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             10912..11092
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11139..11264
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11757..11990
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12175..12219
                     /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"
ORIGIN
        1 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta
       61 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccagac
      121 cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac
      181 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat
      241 tttcgggttt attacaggga cagcagagat ccactttggc gccggctcga gtggctccgg
      301 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
      361 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
      421 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
      481 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtgtcgt gacgcgggat
      541 ccgccaccat ggactataag gaccacgacg gagactacaa ggatcatgat attgattaca
      601 aagacgatga cgataagatg gccccaaaga agaagcggaa ggtcggtatc cacggagtcc
      661 cagcagccat gagctcagag actggcccag tggctgtgga ccccacattg agacggcgga
      721 tcgagcccca tgagtttgag gtattcttcg atccgagaga gctccgcaag gagacctgcc
      781 tgctttacga aattaattgg gggggccggc actccatttg gcgacataca tcacagaaca
      841 ctaacaagca cgtcgaagtc aacttcatcg agaagttcac gacagaaaga tatttctgtc
      901 cgaacacaag gtgcagcatt acctggtttc tcagctggag cccatgcggc gaatgtagta
      961 gggccatcac tgaattcctg tcaaggtatc cccacgtcac tctgtttatt tacatcgcaa
     1021 ggctgtacca ccacgctgac ccccgcaatc gacaaggcct gcgggatttg atctcttcag
     1081 gtgtgactat ccaaattatg actgagcagg agtcaggata ctgctggaga aactttgtga
     1141 attatagccc gagtaatgaa gcccactggc ctaggtatcc ccatctgtgg gtacgactgt
     1201 acgttcttga actgtactgc atcatactgg gcctgcctcc ttgtctcaac attctgagaa
     1261 ggaagcagcc acagctgaca ttctttacca tcgctcttca gtcttgtcat taccagcgac
     1321 tgcccccaca cattctctgg gccaccgggt tgaaaagcgg cagcgagact cccccaaaga
     1381 agaaacggaa agtaggcggc tcccccaaga agaagcggaa ggtagggacc tcagagtccg
     1441 ccacacccga aagtgacaag aagtacagca tcggcctggc catcggcacc aactctgtgg
     1501 gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca
     1561 acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg
     1621 aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga
     1681 accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct
     1741 tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc
     1801 ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc
     1861 acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg
     1921 ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg
     1981 acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg
     2041 aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga
     2101 gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc
     2161 tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg
     2221 acctggccga ggataccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca
     2281 acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt
     2341 ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc
     2401 tgagcgcctc tatgatcaag ctgtacgacg agcaccacca ggacctgacc ctgctgaaag
     2461 ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga
     2521 acggctacgc cggctacatt gacggcggag ccagccagga agagttctac aagttcatca
     2581 agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg
     2641 acctgctgcg gaagcagcgg accttcgaca acggcatcat cccccaccag atccacctgg
     2701 gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc
     2761 gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca
     2821 ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga
     2881 acttcgagaa ggtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca
     2941 acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt
     3001 acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc
     3061 ccgccttcct gagcggcgac cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc
     3121 ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact
     3181 ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc
     3241 tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg
     3301 aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga
     3361 aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca
     3421 ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca
     3481 agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atccagctga
     3541 tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg
     3601 gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca
     3661 tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg
     3721 agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca
     3781 gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga
     3841 aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc
     3901 agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg
     3961 atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc
     4021 tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga
     4081 agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt
     4141 tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca
     4201 tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact
     4261 cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca
     4321 ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg
     4381 agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc
     4441 tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg
     4501 acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact
     4561 tcttctacag caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga
     4621 tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg
     4681 gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa
     4741 agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg
     4801 ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc
     4861 ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac
     4921 tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga
     4981 atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca
     5041 agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg
     5101 ccggcgtgct gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt
     5161 acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc
     5221 tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct
     5281 ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc
     5341 accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca
     5401 atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca
     5461 ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg
     5521 agacacggat cgacctgtct cagctgggag gcgattcagg cggatctact aatctgtcag
     5581 atattattga aaaggagacc ggtaagcaac tggttatcca ggaatccatc ctcatgctcc
     5641 cagaggaggt ggaagaagtc attgggaaca agccggaaag cgatatactc gtgcacaccg
     5701 cctacgacga gagcaccgac gagaatgtca tgcttctgac tagcgacgcc cctgaataca
     5761 agccttgggc tctggtcata caggatagca acggtgagaa caagattaag atgctctctg
     5821 gtggttctcc caagaagaag aggaaagtcg ctagcggcag cggcgccacc aacttcagcc
     5881 tgctgaagca ggccggcgac gtggaggaga accccggccc catgaccgag tacaagccca
     5941 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     6001 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     6061 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     6121 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     6181 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     6241 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     6301 ccgtcggagt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     6361 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     6421 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     6481 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg aacgcgttaa gtcgacaatc
     6541 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     6601 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     6661 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     6721 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     6781 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     6841 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     6901 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     6961 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     7021 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     7081 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcgtcga ctttaagacc
     7141 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     7201 agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg
     7261 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     7321 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     7381 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtacgtatag
     7441 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag
     7501 tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     7561 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     7621 tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat cccgccccta
     7681 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     7741 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     7801 ggcctaggga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
     7861 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     7921 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     7981 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     8041 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     8101 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     8161 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     8221 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     8281 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     8341 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     8401 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     8461 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     8521 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     8581 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     8641 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     8701 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     8761 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     8821 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     8881 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     8941 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     9001 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     9061 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     9121 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     9181 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     9241 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     9301 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     9361 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     9421 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     9481 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     9541 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     9601 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttttgcgcg taatctgctg
     9661 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     9721 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     9781 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     9841 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     9901 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     9961 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
    10021 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
    10081 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
    10141 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
    10201 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
    10261 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
    10321 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
    10381 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
    10441 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
    10501 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
    10561 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
    10621 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgcaagc
    10681 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
    10741 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
    10801 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
    10861 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctt
    10921 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
    10981 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
    11041 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
    11101 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
    11161 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
    11221 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
    11281 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
    11341 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
    11401 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
    11461 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
    11521 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
    11581 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
    11641 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
    11701 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
    11761 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
    11821 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
    11881 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
    11941 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
    12001 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
    12061 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
    12121 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
    12181 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
    12241 aattggctgt ggtatataaa attattcata atgatag
//
