LOCUS       sequence-318793-       14948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of hyA3A-BE4max.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14948)
  AUTHORS   Zhang X, Chen L, Zhu B, Wang L, Chen C, Hong M, Huang Y, Li H, Han 
            H, Cai B, Yu W, Yin S, Yang L, Yang Z, Liu M, Zhang Y, Mao Z, Wu Y, 
            Liu M, Li D
  TITLE     Increasing the efficiency and targeting range of cytidine base 
            editors through fusion of a single-stranded DNA-binding protein 
            domain.
  JOURNAL   Nat Cell Biol. 2020 Jun;22(6):740-750. doi: 
            10.1038/s41556-020-0518-8. Epub 2020 May 11.
  PUBMED    32393889
REFERENCE   2  (bases 1 to 14948)
  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"; doi: 
            "10.1038/s41556-020-0518-8"; journalName: "Nat Cell Biol"; date: 
            "2020-06"; volume: "22"; issue: "6"; pages: "740-750"
COMMENT     Sequence Label: Lenti-117G-hyA3A-BE4max
FEATURES             Location/Qualifiers
     source          1..14948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..200
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             218..398
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    445..570
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1063..1296
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1481..1525
                     /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    1823..1940
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1991..2239
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1991..2011
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2162..2181
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2261..2336
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2393..2604
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2670..2690
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3819..3842,3843..3845,3846..7919)
                     /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             3819..7919
                     /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             7920..7967
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7983..8039
                     /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             8040..8288
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             8319..8567
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             8610..8630
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8640..8696
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(8697..8699,8700..8702,8703..9416)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(8697..8699,8700..8702,8703..9413)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(9003..9022)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9350..9371
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    9423..10011
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9476..9496)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9894..9905)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             10083..10316
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(10342..10359)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10348..10572
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10618..11046
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10705..10724)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10915..10936
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11055..11075)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11060..11389
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11240..11375
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11302..11321
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11437..11484
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11503..11877
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    12007..12128
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12044..12063)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12098..12117
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12177..12193)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12177..12193)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12190..12212)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12201..12217
                     /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)."
     promoter        complement(join(12225..12231,12232..12249,12250..12255))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12270..12291
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12408..12425)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12579..13167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12659..12678)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13338..14129,14130..14198))
                     /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     13961..13980
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14199..14303)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14378..14397)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        14569..14948
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat
      241 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
      301 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
      361 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
      421 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
      481 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
      541 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
      601 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
      661 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
      721 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
      781 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
      841 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
      901 cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
      961 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1021 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1081 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1141 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1201 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1261 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1321 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1381 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1441 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1501 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1561 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1621 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1681 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1741 cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa ttctgcagac
     1801 aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg tacagtgcag
     1861 gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta caaaaacaaa
     1921 ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca gtttggttaa
     1981 ttaaggtacc gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc
     2041 tgttagagag ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac
     2101 gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat
     2161 ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt
     2221 gtggaaagga cgaaacaccg gctattggtc aaggcaaggc gttttagagc tagaaatagc
     2281 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     2341 tttgctagct aggtcttgaa aggagtggga attggctccg gtgcccgtca gtgggcagag
     2401 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg atccggtgcc
     2461 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
     2521 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
     2581 aacgggtttg ccgccagaac acaggggcgc gcctctagag cgatcgcgcc accatgaaga
     2641 ggaccgccga tggctctgag ttcgagagcc ccaagaagaa gcggaaggtg gaggcatctc
     2701 cagcaagcgg accaaggcac ctgatggacc cccacatctt cacctctaac tttaacaatg
     2761 gcatcggcag gcacaagaca tacctgtgct atgaggtgga gcgcctggac aacggcacca
     2821 gcgtgaagat ggatcagcac agaggcttcc tgcacaacca ggccaagaat ctgctgtgcg
     2881 gcttctacgg ccggcacgca gagctgagat ttctggacct ggtgcctagc ctgcagctgg
     2941 atccagccca gatctatagg gtgacctggt tcatcagctg gtccccatgc ttttcctggg
     3001 gatgtgcagg agaggtgcgc gccttcctgc aggagaatac acacgtgcgg ctgagaatct
     3061 ttgccgcccg gatctacgac tatgatcctc tgtacaagga ggccctgcag atgctgagag
     3121 acgcaggagc ccaggtgtcc atcatgacct atgatgagtt caagcactgc tgggacacat
     3181 ttgtggatca ccagggctgt ccctttcagc cttgggacgg actggatgag cactcccagg
     3241 ccctgtctgg caggctgagg gccatcctgc agaaccaggg caatagcgga ggatctagcg
     3301 gaggatcaag cggaagcgag actcctggaa ccagcgaaag cgcaacccca gaaagcagcg
     3361 gaggaagtag cggaggaagc atggcaatgc agatgcagct tgaagcaaat gcagatactt
     3421 cagtggaaga agaaagcttt ggcccacaac ccatttcacg gttagagcag tgtggcataa
     3481 atgccaacga tgtgaagaaa ttggaagaag ctggattcca tactgtggag gctgttgcct
     3541 atgcgccaaa gaaggagcta ataaatatta agggaattag tgaagccaaa gctgataaaa
     3601 ttctggctga ggcagctaaa ttagttccaa tgggtttcac cactgcaact gaattccacc
     3661 aaaggcggtc agagatcata cagattacta ctggctccaa agagcttgac aaactacttc
     3721 aatctggcgg atctagcggt ggatctagcg gctctgagac ccctggaaca tccgaatccg
     3781 ccactccaga gagcagcgga ggctcttctg gaggatcaga caagaagtac agcatcggcc
     3841 tggccatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca
     3901 gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg
     3961 gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca
     4021 gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg
     4081 agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag
     4141 aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc
     4201 acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg
     4261 ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc
     4321 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
     4381 tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca
     4441 aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc
     4501 tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga
     4561 cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg
     4621 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
     4681 tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga
     4741 acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc
     4801 accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag
     4861 agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc
     4921 aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac
     4981 tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca
     5041 gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt
     5101 tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc
     5161 cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga
     5221 gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc
     5281 agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc
     5341 ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat
     5401 acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg
     5461 tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact
     5521 tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg
     5581 cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca
     5641 atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca
     5701 gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga
     5761 agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg
     5821 gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg
     5881 ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc
     5941 agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg
     6001 gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga
     6061 aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga
     6121 ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca
     6181 aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg
     6241 agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg
     6301 acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctgaagg
     6361 acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca
     6421 acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg
     6481 ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga
     6541 gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa
     6601 agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc
     6661 tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg
     6721 atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc
     6781 tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg
     6841 tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa
     6901 tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg
     6961 agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa
     7021 ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca
     7081 tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt
     7141 ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta
     7201 agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag
     7261 tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca
     7321 tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag
     7381 aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg
     7441 gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc
     7501 cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc
     7561 ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga
     7621 tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca
     7681 aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata
     7741 tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca
     7801 ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc
     7861 accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca
     7921 aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa gaaaaaggga tccggcagcg
     7981 gcgcaacaaa cttctctctg ctgaaacaag ccggagatgt cgaagagaat cctggaccga
     8041 ctaatctgag cgacatcatt gagaaggaga ctgggaaaca gctggtcatt caggagtcca
     8101 tcctgatgct gcctgaggag gtggaggaag tgatcggcaa caagccagag tctgacatcc
     8161 tggtgcacac cgcctacgac gagtccacag atgagaatgt aatgctgctg acctctgacg
     8221 cccccgagta taagccttgg gccctggtca tccaggattc taacggcgag aataagatca
     8281 agatgctgag cggaggatcc ggaggatctg gaggcagcac caacctgtct gacatcatcg
     8341 agaaggagac aggcaagcag ctggtcatcc aggagagcat cctgatgctg cccgaagaag
     8401 tcgaagaagt gatcggaaac aagcctgaga gcgatatcct ggtccatacc gcctacgacg
     8461 agagtaccga cgaaaatgtg atgctgctga catccgacgc cccagagtat aagccctggg
     8521 ctctggtcat ccaggattcc aacggagaga acaaaatcaa aatgctgtct ggcggctcaa
     8581 aaagaaccgc cgacggcagc gaattcgagc ccaagaagaa gaggaaagtc ggaagcggag
     8641 ctactaactt cagcctgctg aagcaggctg gagacgtgga ggagaaccct ggacctatgg
     8701 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     8761 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     8821 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     8881 tgaccaccct gacctatgga gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     8941 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     9001 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     9061 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     9121 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     9181 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     9241 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     9301 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     9361 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaggtta
     9421 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     9481 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     9541 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     9601 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     9661 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     9721 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     9781 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     9841 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     9901 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     9961 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cgtcgacttt
    10021 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
    10081 actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc
    10141 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
    10201 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
    10261 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagggc
    10321 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
    10381 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
    10441 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
    10501 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
    10561 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
    10621 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
    10681 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
    10741 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
    10801 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
    10861 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
    10921 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
    10981 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
    11041 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
    11101 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
    11161 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
    11221 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
    11281 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
    11341 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
    11401 agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag
    11461 tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt
    11521 gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg
    11581 ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg
    11641 accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg
    11701 tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc
    11761 cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc
    11821 gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac
    11881 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
    11941 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
    12001 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
    12061 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
    12121 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
    12181 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
    12241 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
    12301 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
    12361 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
    12421 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
    12481 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
    12541 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
    12601 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
    12661 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
    12721 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
    12781 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
    12841 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
    12901 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
    12961 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
    13021 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
    13081 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
    13141 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
    13201 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
    13261 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
    13321 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
    13381 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
    13441 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
    13501 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
    13561 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
    13621 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
    13681 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
    13741 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
    13801 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
    13861 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
    13921 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
    13981 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
    14041 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
    14101 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
    14161 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
    14221 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
    14281 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
    14341 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
    14401 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
    14461 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
    14521 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat
    14581 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
    14641 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
    14701 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
    14761 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
    14821 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
    14881 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
    14941 ttaccatg
//
