LOCUS       sequence-388885-       14358 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human codon-optimized SpCas9-NG-SsAPOBEC3B and blasticidin
            resistance: EFS promoter-SpCas9-NG-SsAPOBEC3B-NLS-FLAG-P2A-BSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14358)
  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 14358)
  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-NG-SsAPOBEC3B
FEATURES             Location/Qualifiers
     source          1..14358
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    417..538
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(454..473)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     508..527
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(587..603)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(587..603)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(600..622)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    611..627
                     /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(635..641,642..659,660..665))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    680..701
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(818..835)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(989..1577)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1069..1088)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1748..2539,2540..2608))
                     /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     2371..2390
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2609..2713)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2788..2807)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2979..3358
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3360..3558
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3513..3533
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3576..3756
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3803..3928
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4421..4654
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4839..4883
                     /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    5181..5298
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5391..5602
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5611..5620
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6553..10653
                     /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
                     SIRPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARFLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPRAFKYFDTTIDRKVYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             10684..10932
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             10963..11211
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             11212..11232
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11278..11298
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11299..11322
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             11332..11388
                     /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             11389..11787
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    11812..12400
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11865..11885)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(12283..12294)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(12403..12419)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12404..12420)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             12925..13105
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(13131..13148)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    13137..13361
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      13407..13835
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(13494..13513)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     13704..13725
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(13844..13864)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      14009..14143
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     14070..14089
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        14205..14252
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             join(14271..14358,1..287)
                     /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"
ORIGIN
        1 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
       61 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
      121 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
      181 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
      241 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
      301 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
      361 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
      421 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
      481 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
      541 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
      601 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
      661 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
      721 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
      781 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
      841 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
      901 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
      961 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1021 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1081 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1141 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1201 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1261 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1321 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1381 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1441 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1501 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1561 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1621 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1681 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     1741 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     1801 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     1861 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     1921 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     1981 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     2041 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     2101 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     2161 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     2221 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     2281 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     2341 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     2401 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     2461 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     2521 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     2581 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     2641 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2701 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     2761 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     2821 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     2881 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     2941 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
     3001 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     3061 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     3121 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     3181 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     3241 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     3301 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     3361 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     3421 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     3481 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     3541 ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct
     3601 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     3661 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     3721 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     3781 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     3841 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     3901 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     3961 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     4021 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     4081 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     4141 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     4201 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     4261 gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
     4321 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
     4381 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
     4441 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
     4501 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
     4561 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
     4621 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
     4681 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
     4741 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
     4801 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
     4861 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
     4921 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
     4981 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
     5041 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
     5101 gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa
     5161 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
     5221 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     5281 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttggttaatt
     5341 agctagctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg
     5401 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgat ccggtgccta
     5461 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
     5521 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
     5581 cgggtttgcc gccagaacac aggaccggtt gccaccatgg acccacagag gctgcgccag
     5641 tggcccggcc ctggcccagc aagcaggggc ggctacggcc agcggccaag aatcaggaac
     5701 cccgaggagt ggtttcacga gctgtctccc cggaccttca gctttcactt ccgcaacctg
     5761 aggttcgcat ccggccgcaa tcggtcttat atctgctgtc aggtggaggg caagaactgc
     5821 ttctttcagg gcatctttca gaatcaggtg ccacctgacc caccatgcca cgcagagctg
     5881 tgcttcctgt cttggttcca gagctggggc ctgtcccccg atgagcacta ctatgtgaca
     5941 tggtttatct cttggagccc ttgctgtgag tgtgccgcca aggtggccca gttcctggag
     6001 gagaaccgca acgtgagcct gtctctgagc gccgcaaggc tgtactattt ctggaagtcc
     6061 gagtctagag agggactgcg gagactgagc gacctgggag cacaagtggg aatcatgtcc
     6121 tttcaggatt tccagcactg ctggaacaat tttgtgcaca acctgggcat gcccttccag
     6181 ccttggaaga agctgcacaa gaattaccag aggctggtga ccgagctgaa gcagatcctg
     6241 cgcgaggagc ctgccacata tggctctcca caggcccagg gcaaggtgag aatcggaagc
     6301 accgcagcag gactgaggca cagccactcc cacacacgct ccgaggcaca cctgaggcct
     6361 aaccacagct ccagacagca caggatcctg aatcctccac gggaggccag agccaggacc
     6421 tgcgtgctgg tggatgcctc ttggatctgt tacagaagcg gaggatctag cggcggctct
     6481 agcggatctg agacacctgg cacaagcgag tctgccacac ctgagagtag cggcggatct
     6541 tctggtggct ctgacaagaa gtacagcatc ggcctggcca tcggcaccaa ctctgtgggc
     6601 tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac
     6661 accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa
     6721 acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac
     6781 cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc
     6841 ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc
     6901 atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac
     6961 ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc
     7021 ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac
     7081 aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag
     7141 gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc
     7201 aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg
     7261 ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac
     7321 ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac
     7381 ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc
     7441 gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg
     7501 agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct
     7561 ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac
     7621 ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag
     7681 cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac
     7741 ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga
     7801 gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg
     7861 gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg
     7921 ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac
     7981 ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac
     8041 ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac
     8101 ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc
     8161 gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg
     8221 aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc
     8281 gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg
     8341 ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa
     8401 gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa
     8461 acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc
     8521 ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag
     8581 acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc
     8641 cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc
     8701 gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc
     8761 ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag
     8821 aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc
     8881 cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa
     8941 gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag
     9001 aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat
     9061 gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg
     9121 accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag
     9181 aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc
     9241 gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc
     9301 aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc
     9361 cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc
     9421 ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag
     9481 atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg
     9541 atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac
     9601 gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc
     9661 ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc
     9721 cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc
     9781 cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag
     9841 accgaggtgc agacaggcgg cttcagcaaa gagtctatcc ggcccaagag gaacagcgat
     9901 aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt cgtgagcccc
     9961 accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg
    10021 aagagtgtga aagagctact ggggatcacc atcatggaaa gaagcagctt cgagaagaat
    10081 cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag
    10141 ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc
    10201 cggttcctgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac
    10261 ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg
    10321 tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc
    10381 aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac
    10441 cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat
    10501 ctgggagccc ctcgggcctt caagtacttt gacaccacca tcgaccggaa ggtgtaccgg
    10561 agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag
    10621 acacggatcg acctgtctca gctgggaggc gactctggtg gaagcggagg atctggcggc
    10681 agcaccaatc tgagcgacat catcgagaaa gagacaggca agcagctggt catccaagag
    10741 tccatcctga tgctgcctga agaggtggaa gaagtgatcg gcaacaagcc cgagtccgac
    10801 atcctggtgc acaccgccta cgatgagagc accgacgaga acgtgatgct gctgacctct
    10861 gacgcccctg agtacaagcc ttgggctctc gtgatccagg acagcaacgg cgagaacaag
    10921 atcaagatgc tgagcggcgg ctctggtggc tctggcggat ctacaaacct gtccgatatt
    10981 attgagaaag aaaccgggaa acagctcgtg attcaagagt ctattctcat gctcccggaa
    11041 gaagtcgagg aagtcattgg aaacaagcct gagagcgata ttctggtcca tacagcctac
    11101 gacgagtcta ccgatgagaa tgtcatgctc ctcaccagcg acgctcccga gtataagcca
    11161 tgggcacttg tcattcagga ctccaatggg gaaaacaaaa tcaaaatgct cccaaagaaa
    11221 aaacgcaagg tggagggagc tgataagcgc accgccgatg gttccgagtt cgaaagcccc
    11281 aagaagaaga ggaaagtcga ttacaaagac gatgacgata agggatccgg cgcaacaaac
    11341 ttctctctgc tgaaacaagc cggagatgtc gaagagaatc ctggaccgat ggccaagcct
    11401 ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc
    11461 atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact
    11521 ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact
    11581 gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg
    11641 ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc
    11701 aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg
    11761 ccctctggtt atgtgtggga gggctaagaa ttcgatatca agcttatcgg taatcaacct
    11821 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
    11881 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
    11941 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
    12001 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccgcac tggttggggc
    12061 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
    12121 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
    12181 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
    12241 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
    12301 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
    12361 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga
    12421 gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt
    12481 gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct
    12541 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
    12601 ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac
    12661 cacacacaag gctacttccc tgattggcag aactacacac cagggccagg gatcagatat
    12721 ccactgacct ttggatggtg ctacaagcta gtaccagttg agcaagagaa ggtagaagaa
    12781 gccaatgaag gagagaacac ccgcttgtta caccctgtga gcctgcatgg gatggatgac
    12841 ccggagagag aagtattaga gtggaggttt gacagccgcc tagcatttca tcacatggcc
    12901 cgagagctgc atccggactg tactgggtct ctctggttag accagatctg agcctgggag
    12961 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
    13021 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
    13081 tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag cctcgactgt
    13141 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
    13201 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
    13261 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
    13321 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
    13381 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
    13441 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
    13501 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
    13561 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
    13621 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
    13681 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
    13741 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
    13801 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
    13861 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
    13921 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
    13981 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccag ttccgcccat
    14041 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
    14101 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
    14161 ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat
    14221 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt
    14281 tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga
    14341 ccgaccggct cgggttct
//
