LOCUS       sequence-390295-       14235 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized SpCas9-YE1-BE4max and blasticidin 
            resistance: EFS promoter-SpCas9-YE1-BE4max-NLS-FLAG-P2A-BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14235)
  AUTHORS   Kim N, Choi S, Kim S, Song M, Seo JH, Min S, Park J, Cho SR, Kim HH
  TITLE     Deep learning models to predict the editing efficiencies and 
            outcomes of diverse base editors.
  JOURNAL   Nat Biotechnol. 2024 Mar;42(3):484-497. doi: 
            10.1038/s41587-023-01792-x
  PUBMED    37188916
REFERENCE   2  (bases 1 to 14235)
  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"; journalName: 
            "Nat Biotechnol"; date: "2024-03"; volume: "42"; issue: "3"; pages: 
            "484-497"
COMMENT     Sequence Label: SpCas9-YE1-BE4max
FEATURES             Location/Qualifiers
     source          1..14235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        100..479
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        481..679
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     634..654
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             697..877
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    924..1049
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1542..1775
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1960..2004
                     /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    2302..2419
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2512..2723
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2774..2794
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3575..3598,3599..3601,3602..7675)
                     /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             3575..7675
                     /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             7706..7954
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             7985..8233
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             8276..8296
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8297..8320
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8330..8386
                     /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             8387..8785
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8810..9398
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8863..8883)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9281..9292)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9401..9417)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9402..9418)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9923..10103
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10129..10146)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10135..10359
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10405..10833
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10492..10511)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10702..10723
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10842..10862)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      11007..11141
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11068..11087
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11203..11250
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11269..11643
                     /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    11773..11894
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11810..11829)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11864..11883
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11943..11959)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11943..11959)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11956..11978)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11967..11983
                     /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(11991..11997,11998..12015,12016..12021))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12036..12057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12174..12191)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12345..12933)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12425..12444)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13104..13895,13896..13964))
                     /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     13727..13746
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13965..14069)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14144..14163)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
       61 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
      121 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      181 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      241 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      301 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      361 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      421 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      481 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      541 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      601 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      661 gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt agaccagatc
      721 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
      781 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
      841 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
      901 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
      961 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
     1021 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
     1081 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
     1141 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
     1201 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
     1261 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
     1321 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
     1381 agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
     1441 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     1501 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     1561 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     1621 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     1681 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     1741 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     1801 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     1861 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     1921 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     1981 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     2041 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     2101 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     2161 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     2221 agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat tctgcagaca
     2281 aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt acagtgcagg
     2341 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     2401 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag tttggttaat
     2461 tagctagcta ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc
     2521 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga tccggtgcct
     2581 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
     2641 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
     2701 acgggtttgc cgccagaaca caggaccggt tgccaccatg aaacggacag ccgacggaag
     2761 cgagttcgag tcaccaaaga agaagcggaa agtctcctca gagactgggc ctgtcgccgt
     2821 cgatccaacc ctgcgccgcc ggattgaacc tcacgagttt gaagtgttct ttgacccccg
     2881 ggagctgaga aaggagacat gcctgctgta cgagatcaac tggggaggca ggcactccat
     2941 ctggaggcac acctctcaga acacaaataa gcacgtggag gtgaacttca tcgagaagtt
     3001 taccacagag cggtacttct gccccaatac cagatgtagc atcacatggt ttctgagcta
     3061 ttccccttgc ggagagtgta gcagggccat caccgagttc ctgtccagat atccacacgt
     3121 gacactgttt atctacatcg ccaggctgta tcaccacgca gacccagaga ataggcaggg
     3181 cctgcgcgat ctgatcagct ccggcgtgac catccagatc atgacagagc aggagtccgg
     3241 ctactgctgg cggaacttcg tgaattattc tcctagcaac gaggcccact ggcctaggta
     3301 cccacacctg tgggtgcgcc tgtacgtgct ggagctgtat tgcatcatcc tgggcctgcc
     3361 cccttgtctg aatatcctgc ggagaaagca gccccagctg accttcttta caatcgccct
     3421 gcagtcttgt cactatcaga ggctgccacc ccacatcctg tgggccacag gcctgaagtc
     3481 tggaggatct agcggaggat cctctggcag cgagacacca ggaacaagcg agtcagcaac
     3541 accagagagc agtggcggca gcagcggcgg cagcgacaag aagtacagca tcggcctggc
     3601 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
     3661 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc
     3721 cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag
     3781 aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat
     3841 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
     3901 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
     3961 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
     4021 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
     4081 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
     4141 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
     4201 catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc
     4261 cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc
     4321 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
     4381 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
     4441 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
     4501 cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca
     4561 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
     4621 tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga
     4681 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
     4741 cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat
     4801 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta
     4861 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
     4921 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
     4981 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag
     5041 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
     5101 gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt
     5161 gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga
     5221 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
     5281 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
     5341 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
     5401 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
     5461 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
     5521 gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
     5581 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
     5641 cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa
     5701 agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag
     5761 ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt
     5821 gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac
     5881 ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga
     5941 gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa
     6001 gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat
     6061 caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga
     6121 ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt
     6181 gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa
     6241 gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga
     6301 actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca
     6361 cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat
     6421 ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt
     6481 ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa
     6541 cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta
     6601 cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg
     6661 caaggctacc gccaagtact tcttctacag caacatcatg aactttttca agaccgagat
     6721 taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg
     6781 ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc
     6841 ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat
     6901 cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa
     6961 gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga
     7021 aaagggcaag tccaagaaac tgaagagtgt gaaagagcta ctggggatca ccatcatgga
     7081 aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt
     7141 gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg
     7201 gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc
     7261 caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga
     7321 ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat
     7381 cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt
     7441 gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat
     7501 ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac
     7561 catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca
     7621 gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacagcgg
     7681 cgggagcggc gggagcgggg ggagcactaa tctgagcgac atcattgaga aggagactgg
     7741 gaaacagctg gtcattcagg agtccatcct gatgctgcct gaggaggtgg aggaagtgat
     7801 cggcaacaag ccagagtctg acatcctggt gcacaccgcc tacgacgagt ccacagatga
     7861 gaatgtgatg ctgctgacct ctgacgcccc cgagtataag ccttgggccc tggtcatcca
     7921 ggattctaac ggcgagaata agatcaagat gctgagcgga ggatccggag gatctggagg
     7981 cagcaccaac ctgtctgaca tcatcgagaa ggagacaggc aagcagctgg tcatccagga
     8041 gagcatcctg atgctgcccg aagaagtcga agaagtgatc ggaaacaagc ctgagagcga
     8101 tatcctggtc cataccgcct acgacgagag taccgacgaa aatgtgatgc tgctgacatc
     8161 cgacgcccca gagtataagc cctgggctct ggtcatccag gattccaacg gagagaacaa
     8221 aatcaaaatg ctgtctggcg gctcaaaaag aaccgccgac ggcagcgaat tcgagcccaa
     8281 gaagaagagg aaagtcgatt acaaagacga tgacgataag ggatccggcg caacaaactt
     8341 ctctctgctg aaacaagccg gagatgtcga agagaatcct ggaccgatgg ccaagccttt
     8401 gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat
     8461 ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg
     8521 tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc
     8581 tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg
     8641 catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa
     8701 agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc
     8761 ctctggttat gtgtgggagg gctaagaatt cgatatcaag cttatcggta atcaacctct
     8821 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     8881 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     8941 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     9001 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     9061 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     9121 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     9181 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     9241 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     9301 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     9361 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc gacctcgaga
     9421 cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc
     9481 ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt
     9541 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
     9601 actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca
     9661 cacacaaggc tacttccctg attggcagaa ctacacacca gggccaggga tcagatatcc
     9721 actgaccttt ggatggtgct acaagctagt accagttgag caagagaagg tagaagaagc
     9781 caatgaagga gagaacaccc gcttgttaca ccctgtgagc ctgcatggga tggatgaccc
     9841 ggagagagaa gtattagagt ggaggtttga cagccgccta gcatttcatc acatggcccg
     9901 agagctgcat ccggactgta ctgggtctct ctggttagac cagatctgag cctgggagct
     9961 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
    10021 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
    10081 gtcagtgtgg aaaatctcta gcagggcccg tttaaacccg ctgatcagcc tcgactgtgc
    10141 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
    10201 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
    10261 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
    10321 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
    10381 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
    10441 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
    10501 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
    10561 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
    10621 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
    10681 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
    10741 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
    10801 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
    10861 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
    10921 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
    10981 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccagtt ccgcccattc
    11041 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
    11101 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
    11161 cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg
    11221 gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg
    11281 accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc
    11341 gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac
    11401 gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc
    11461 tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg
    11521 aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg
    11581 gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac
    11641 tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
    11701 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
    11761 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
    11821 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
    11881 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
    11941 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
    12001 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
    12061 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
    12121 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
    12181 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
    12241 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
    12301 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
    12361 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
    12421 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
    12481 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
    12541 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
    12601 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
    12661 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
    12721 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
    12781 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
    12841 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
    12901 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
    12961 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
    13021 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
    13081 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
    13141 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
    13201 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
    13261 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
    13321 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
    13381 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
    13441 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
    13501 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
    13561 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
    13621 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
    13681 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
    13741 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
    13801 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
    13861 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
    13921 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
    13981 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
    14041 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
    14101 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
    14161 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
    14221 cgagcaaaat ttaag
//
