LOCUS       sequence-458310-       14736 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ef1a-PmCDA1-BE3-T2A-BlastR-wPRE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14736)
  AUTHORS   Schwartz CI, Abell NS, Li A, Aradhana, Tycko J, Truong A, Montgomery
            SB, Hess GT
  TITLE     Toward optimizing diversifying base editors for high-throughput 
            mutational scanning studies.
  JOURNAL   Nucleic Acids Res. 2025 Jun 20;53(12):gkaf620. doi: 
            10.1093/nar/gkaf620.
  PUBMED    40613705
REFERENCE   2  (bases 1 to 14736)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2025-06-20"; volume: "53(12):gkaf620. doi"; 
            pages: " 10.1093/nar/gkaf620"
COMMENT     Sequence Label: pGH903
FEATURES             Location/Qualifiers
     source          1..14736
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2687..3865
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2918..3856
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3813..3833
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3878..4501
                     /codon_start=1
                     /product="cytidine deaminase from the sea lamprey 
                     Petromyzon marinus (Rogozin et al., 2007)"
                     /label=PmCDA1
                     /translation="MTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGER
                     RACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKI
                     LEWYNQELRGNGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIF
                     IQSSHNQLNENRWLEKTLKRAEKRRSELSIMIQVKILHTTKSPAV"
     CDS             join(4550..4573,4574..4576,4577..7132,7133..7135,
                     7136..8650)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     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             join(4550..4573,4574..4576,4577..8650)
                     /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             8663..8911
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             8924..8944
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8951..9004
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             9005..9403
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9428..10016
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9481..9501)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9899..9910)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10019..10035)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10020..10036)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10541..10721
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10747..10764)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10753..10977
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11023..11451
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11110..11129)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11320..11341
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11460..11480)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11465..11794
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11645..11780
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11707..11726
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11842..11889
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11908..12282
                     /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    12412..12533
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12449..12468)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12503..12522
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12582..12598)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12582..12598)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12595..12617)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12606..12622
                     /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(12630..12636,12637..12654,12655..12660))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12675..12696
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12813..12830)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12984..13572)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13064..13083)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13743..14534,14535..14603))
                     /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     14366..14385
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14604..14708)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta gctagctgca aagatggata aagttttaaa cagagaggaa
     2641 tctttgcagc taatggacct tctaggtctt gaaaggagtg ggaattggct ccggtgcccg
     2701 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
     2761 ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
     2821 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
     2881 cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg
     2941 cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct
     3001 gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc
     3061 ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg
     3121 ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct
     3181 agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt
     3241 aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg
     3301 ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag
     3361 aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc
     3421 gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga
     3481 aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg
     3541 agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc
     3601 ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt
     3661 ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac
     3721 tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt
     3781 tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt
     3841 ttttcttcca tttcaggtgt cgtgacgtac ggccaccatg acggatgccg aatatgttcg
     3901 catacatgaa aaattggaca tttatacctt caagaagcag ttcttcaaca ataaaaagtc
     3961 cgtatcacat cgctgctatg tattgtttga actgaagcga aggggtgaaa ggagggcttg
     4021 tttttggggt tatgctgtca acaagccaca gtctgggact gagcgcggca tacatgctga
     4081 gatcttctcc atcagaaaag tcgaagagta tctcagagac aaccctggcc aattcacaat
     4141 aaactggtat tcctcctggt caccatgcgc agattgcgcg gagaaaatac tcgaatggta
     4201 taatcaggaa ctgagaggaa acggacatac tctgaaaatt tgggcgtgca agctgtatta
     4261 tgagaaaaac gcacgaaatc agatcggttt gtggaacctt cgcgataatg gtgtaggcct
     4321 gaacgtcatg gtatctgagc attaccaatg ctgtaggaaa atattcatcc agagcagcca
     4381 caaccagctc aacgaaaatc gctggttgga aaagacactt aaaagagcag aaaagagacg
     4441 cagcgaactc tcaataatga ttcaggtcaa aatattgcat acgacgaaga gtccagccgt
     4501 tagcggcagc gagactcccg ggacctcaga gtccgccaca cccgaaagtg acaagaagta
     4561 cagcatcggc ctggccatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta
     4621 caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa
     4681 gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa
     4741 gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat
     4801 cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt
     4861 cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga
     4921 ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag
     4981 caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg
     5041 gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt
     5101 catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg
     5161 cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct
     5221 gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggcaacctga ttgccctgag
     5281 cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca
     5341 gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca
     5401 gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat
     5461 cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata
     5521 cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga
     5581 gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg
     5641 cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg
     5701 caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt
     5761 cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg
     5821 gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac
     5881 cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat
     5941 gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg
     6001 cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga
     6061 gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac
     6121 caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa
     6181 aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa
     6241 agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga
     6301 tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga
     6361 cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact
     6421 gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga
     6481 caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa
     6541 gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc
     6601 cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa
     6661 agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc
     6721 caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga
     6781 cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag
     6841 agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga
     6901 agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca
     6961 gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga
     7021 ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gaccacatcg tgcctcagag
     7081 ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg
     7141 caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca
     7201 gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag
     7261 aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg
     7321 gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga
     7381 gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga
     7441 tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca
     7501 cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga
     7561 aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag
     7621 cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt
     7681 tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac
     7741 aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa
     7801 agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt
     7861 cagcaaagag tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga
     7921 ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt
     7981 ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg
     8041 gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa
     8101 gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga
     8161 gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga
     8221 actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct
     8281 gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta
     8341 cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc
     8401 taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca
     8461 ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa
     8521 gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc
     8581 caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct
     8641 gggaggcgac agcggtggtt ctactaatct gtcagatatt attgaaaagg agaccggtaa
     8701 gcaactggtt atccaggaat ccatcctcat gctcccagag gaggtggaag aagtcattgg
     8761 gaacaagccg gaaagcgata tactcgtgca caccgcctac gacgagagca ccgacgagaa
     8821 tgtcatgctt ctgactagcg acgcccctga atacaagcct tgggctctgg tcatacagga
     8881 tagcaacggt gagaacaaga ttaagatgct ctctggtggt tctcccaaga agaagaggaa
     8941 agtcggatcc gagggcagag gaagtctgct aacatgcggt gacgtcgagg agaatcctgg
     9001 cccaatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag caacggctac
     9061 aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc tctctagcga
     9121 cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt gtgcagaact
     9181 cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat
     9241 cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg tgcttctcga
     9301 tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga cggcagttgg
     9361 gattcgtgaa ttgctgccct ctggttatgt gtgggagggc taagaattcg atatcaagct
     9421 tatcggtaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     9481 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     9541 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     9601 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     9661 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     9721 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     9781 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     9841 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     9901 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     9961 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
    10021 ataccgtcga cctcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc
    10081 taccaatgct gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt
    10141 cacacctcag gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
    10201 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatatcct
    10261 tgatctgtgg atctaccaca cacaaggcta cttccctgat tggcagaact acacaccagg
    10321 gccagggatc agatatccac tgacctttgg atggtgctac aagctagtac cagttgagca
    10381 agagaaggta gaagaagcca atgaaggaga gaacacccgc ttgttacacc ctgtgagcct
    10441 gcatgggatg gatgacccgg agagagaagt attagagtgg aggtttgaca gccgcctagc
    10501 atttcatcac atggcccgag agctgcatcc ggactgtact gggtctctct ggttagacca
    10561 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
    10621 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
    10681 atccctcaga cccttttagt cagtgtggaa aatctctagc agggcccgtt taaacccgct
    10741 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
    10801 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
    10861 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
    10921 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
    10981 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
    11041 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
    11101 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
    11161 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
    11221 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
    11281 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
    11341 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
    11401 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
    11461 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
    11521 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
    11581 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
    11641 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
    11701 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
    11761 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
    11821 tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta
    11881 taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc
    11941 accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg
    12001 gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc
    12061 gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg
    12121 gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg
    12181 ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg
    12241 gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc
    12301 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
    12361 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
    12421 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
    12481 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
    12541 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
    12601 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
    12661 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
    12721 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
    12781 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
    12841 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
    12901 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
    12961 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
    13021 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
    13081 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
    13141 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
    13201 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
    13261 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
    13321 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
    13381 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
    13441 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
    13501 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
    13561 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
    13621 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
    13681 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
    13741 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
    13801 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
    13861 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
    13921 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
    13981 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
    14041 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
    14101 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
    14161 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
    14221 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
    14281 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
    14341 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
    14401 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
    14461 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
    14521 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
    14581 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
    14641 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
    14701 gttccgcgca catttccccg aaaagtgcca cctgac
//
