LOCUS       sequence-211403-       12157 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector for constitutive expression of 2X in mammalian 
            cells (codon optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12157)
  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 12157)
  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-2X-P2A-Puro
FEATURES             Location/Qualifiers
     source          1..12157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..253
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             254..434
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    481..606
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1099..1332
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1517..1561
                     /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    1745..1862
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1931..2142
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2168..2854
                     /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             2873..2893
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2903..2923
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2954..2977,2978..2980,2981..7054)
                     /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             2954..7054
                     /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             7067..7315
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7328..7348
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7364..7420
                     /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             7421..8020
                     /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(7421..7440)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7917..7937
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    8036..8624
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8089..8109)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8507..8518)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8696..8929
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9007..9128
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9044..9063)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9098..9117
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9168..9303
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9323..9342)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9324..9342)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9352..9369)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9352..9368)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9361..9383)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9510..9965
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9597..9616)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9807..9828
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9991..10095
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10096..10164,10165..10956)
                     /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(10314..10333)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11127..11715
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11616..11635
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11869..11886
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12003..12024
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12039..12044,12045..12062,12063..12069)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12077..12093
                     /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     12082..12104
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12101..12117
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12101..12117
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12136..12156
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        12138..12156
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aacaaaagct ggagctgcaa gcttaatgta gtcttatgca atactcttgt agtcttgcaa
       61 catggtaacg atgagttagc aacatgcctt acaaggagag aaaaagcacc gtgcatgccg
      121 attggtggaa gtaaggtggt acgatcgtgc cttattagga aggcaacaga cgggtctgac
      181 atggattgga cgaaccactg aattgccgca ttgcagagat attgtattta agtgcctagc
      241 tcgatacata aacgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
      301 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
      361 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
      421 gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag
      481 ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga
      541 ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag
      601 agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca
      661 gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga
      721 ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag
      781 ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca
      841 accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag
      901 atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag
      961 acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt
     1021 aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga
     1081 aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac
     1141 tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt
     1201 gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac
     1261 agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga
     1321 tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc
     1381 ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat
     1441 ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc
     1501 gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt
     1561 gtggaattgg tttaacataa caaattggct gtggtatata aaattattca taatgatagt
     1621 aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag
     1681 gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg gacccagacc
     1741 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     1801 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     1861 ttcgggttta ttacagggac agcagagatc cactttggcg ccggctcgag tggctccggt
     1921 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     1981 ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     2041 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     2101 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtgtcgtg acgcgggatc
     2161 cgccaccatg agctcagaga ctggcccagt ggctgtggac cccacattga gacggcggat
     2221 cgagccccat gagtttgagg tattcttcga tccgagagag ctccgcaagg agacctgcct
     2281 gctttacgaa attaattggg ggggccggca ctccatttgg cgacatacat cacagaacac
     2341 taacaagcac gtcgaagtca acttcatcga gaagttcacg acagaaagat atttctgtcc
     2401 gaacacaagg tgcagcatta cctggtttct cagctggagc ccatgcggcg aatgtagtag
     2461 ggccatcact gaattcctgt caaggtatcc ccacgtcact ctgtttattt acatcgcaag
     2521 gctgtaccac cacgctgacc cccgcaatcg acaaggcctg cgggatttga tctcttcagg
     2581 tgtgactatc caaattatga ctgagcagga gtcaggatac tgctggagaa actttgtgaa
     2641 ttatagcccg agtaatgaag cccactggcc taggtatccc catctgtggg tacgactgta
     2701 cgttcttgaa ctgtactgca tcatactggg cctgcctcct tgtctcaaca ttctgagaag
     2761 gaagcagcca cagctgacat tctttaccat cgctcttcag tcttgtcatt accagcgact
     2821 gcccccacac attctctggg ccaccgggtt gaaaagcggc agcgagactc ccccaaagaa
     2881 gaaacggaaa gtaggcggct cccccaagaa gaagcggaag gtagggacct cagagtccgc
     2941 cacacccgaa agtgacaaga agtacagcat cggcctggcc atcggcacca actctgtggg
     3001 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
     3061 caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga
     3121 aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa
     3181 ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt
     3241 cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc
     3301 catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca
     3361 cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc
     3421 cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga
     3481 caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga
     3541 ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag
     3601 caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct
     3661 gttcggcaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga
     3721 cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa
     3781 cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc
     3841 cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct
     3901 gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc
     3961 tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa
     4021 cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa
     4081 gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga
     4141 cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg
     4201 agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg
     4261 ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag
     4321 gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa
     4381 cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa
     4441 cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta
     4501 cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc
     4561 cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg
     4621 gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc
     4681 cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct
     4741 gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga
     4801 agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa
     4861 aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac
     4921 cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa
     4981 gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat
     5041 ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg
     5101 cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat
     5161 cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga
     5221 gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag
     5281 ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa
     5341 agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca
     5401 gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga
     5461 tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct
     5521 gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa
     5581 gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt
     5641 cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat
     5701 caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc
     5761 ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac
     5821 cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga
     5881 gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct
     5941 gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga
     6001 cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt
     6061 cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat
     6121 ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg
     6181 ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa
     6241 gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga
     6301 taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc
     6361 caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact
     6421 gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa
     6481 tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa
     6541 gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc
     6601 cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta
     6661 cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct
     6721 gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc
     6781 caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca
     6841 ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa
     6901 tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac
     6961 cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga
     7021 gacacggatc gacctgtctc agctgggagg cgattcaggc ggatctacta atctgtcaga
     7081 tattattgaa aaggagaccg gtaagcaact ggttatccag gaatccatcc tcatgctccc
     7141 agaggaggtg gaagaagtca ttgggaacaa gccggaaagc gatatactcg tgcacaccgc
     7201 ctacgacgag agcaccgacg agaatgtcat gcttctgact agcgacgccc ctgaatacaa
     7261 gccttgggct ctggtcatac aggatagcaa cggtgagaac aagattaaga tgctctctgg
     7321 tggttctccc aagaagaaga ggaaagtcgc tagcggcagc ggcgccacca acttcagcct
     7381 gctgaagcag gccggcgacg tggaggagaa ccccggcccc atgaccgagt acaagcccac
     7441 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     7501 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     7561 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     7621 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     7681 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     7741 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     7801 cgtcggagtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     7861 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     7921 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     7981 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga acgcgttaag tcgacaatca
     8041 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     8101 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     8161 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     8221 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     8281 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     8341 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     8401 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     8461 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     8521 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     8581 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcgtcgac tttaagacca
     8641 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     8701 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg
     8761 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     8821 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     8881 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tacgtatagt
     8941 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     9001 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     9061 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     9121 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc ccgcccctaa
     9181 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     9241 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     9301 gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt
     9361 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     9421 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     9481 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     9541 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     9601 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     9661 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     9721 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     9781 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     9841 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     9901 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     9961 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
    10021 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
    10081 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
    10141 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
    10201 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
    10261 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
    10321 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
    10381 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
    10441 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
    10501 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
    10561 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
    10621 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
    10681 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
    10741 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
    10801 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
    10861 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
    10921 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
    10981 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
    11041 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
    11101 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
    11161 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
    11221 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
    11281 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
    11341 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
    11401 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
    11461 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
    11521 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
    11581 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
    11641 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
    11701 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
    11761 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
    11821 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
    11881 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
    11941 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
    12001 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
    12061 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
    12121 gattacgcca agcgcgcaat taaccctcac taaaggg
//
