LOCUS       sequence-270570-       12010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline inducible piggyBAC plasmid for mammalian expression of 
            cytosine base editor AncBE4max.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12010)
  AUTHORS   McGrath E, Shin H, Zhang L, Phue JN, Wu WW, Shen RF, Jang YY, 
            Revollo J, Ye Z
  TITLE     Targeting specificity of APOBEC-based cytosine base editor in human 
            iPSCs determined by whole genome sequencing.
  JOURNAL   Nat Commun. 2019 Nov 25;10(1):5353. doi: 10.1038/s41467-019-13342-8.
  PUBMED    31767844
REFERENCE   2  (bases 1 to 12010)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2019-11-25"; volume: "10(1):5353. doi"; pages: " 
            10.1038/s41467-019-13342-8"
COMMENT     Sequence Label: PB-XL-AncBE4max
FEATURES             Location/Qualifiers
     source          1..12010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..368
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     protein_bind    9..27
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    45..63
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    81..99
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    117..135
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    153..171
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    189..207
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    225..243
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             417..437
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1218..1241,1242..1244,1245..5318)
                     /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             1218..5318
                     /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             5349..5597
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5628..5876
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5919..5939
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5949..6005
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(6006..6008,6009..6011,6012..6722)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6006..6008,6009..6011,6012..6722)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6312..6331)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6659..6680
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6735..6755
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6764..6781
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    6964..7098
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   7374..7408
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7419..7436)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7419..7435)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7428..7450)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7644..7663)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7763..7785
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7823..7841)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7909..8013
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8014..8082,8083..8874)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8232..8251)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9045..9633
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9534..9553
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9787..9804
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9921..9942
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9957..9962,9963..9980,9981..9987)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9995..10011
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     10000..10022
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10019..10035
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10019..10035
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   10061..10123
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    10299..10380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(10497..10895)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(10896..10952)
                     /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             complement(10962..11705)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             complement(11706..11729)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      11729..11738
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(11758..11969)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
ORIGIN
        1 gagtttactc cctatcagtg atagagaacg tatgaagagt ttactcccta tcagtgatag
       61 agaacgtatg cagactttac tccctatcag tgatagagaa cgtataagga gtttactccc
      121 tatcagtgat agagaacgta tgaccagttt actccctatc agtgatagag aacgtatcta
      181 cagtttactc cctatcagtg atagagaacg tatatccagt ttactcccta tcagtgatag
      241 agaacgtata agctttgctt atgtaaacca gggcgcctat aaaagagtgc tgattttttg
      301 agtaaacttc aattccacaa cacttttgtc ttataccaac tttccgtacc acttcctacc
      361 ctcgtaaagg taccgccacc atgaaacgga cagccgacgg aagcgagttc gagtcaccaa
      421 agaagaagcg gaaagtcagc agtgaaaccg gaccagtggc agtggaccca accctgagga
      481 gacggattga gccccatgaa tttgaagtgt tctttgaccc aagggagctg aggaaggaga
      541 catgcctgct gtacgagatc aagtggggca caagccacaa gatctggcgc cacagctcca
      601 agaacaccac aaagcacgtg gaagtgaatt tcatcgagaa gtttacctcc gagcggcact
      661 tctgcccctc taccagctgt tccatcacat ggtttctgtc ttggagccct tgcggcgagt
      721 gttccaaggc catcaccgag ttcctgtctc agcaccctaa cgtgaccctg gtcatctacg
      781 tggcccggct gtatcaccac atggaccagc agaacaggca gggcctgcgc gatctggtga
      841 attctggcgt gaccatccag atcatgacag ccccagagta cgactattgc tggcggaact
      901 tcgtgaatta tccacctggc aaggaggcac actggccaag atacccaccc ctgtggatga
      961 agctgtatgc actggagctg cacgcaggaa tcctgggcct gcctccatgt ctgaatatcc
     1021 tgcggagaaa gcagccccag ctgacatttt tcaccattgc tctgcagtct tgtcactatc
     1081 agcggctgcc tcctcatatt ctgtgggcta caggcctgaa gtctggagga tctagcggag
     1141 gatcctctgg cagcgagaca ccaggaacaa gcgagtcagc aacaccagag agcagtggcg
     1201 gcagcagcgg cggcagcgac aagaagtaca gcatcggcct ggccatcggc accaactctg
     1261 tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg
     1321 gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg
     1381 gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga
     1441 agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca
     1501 gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc
     1561 accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct
     1621 accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc
     1681 tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc
     1741 ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt
     1801 tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac
     1861 tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg
     1921 gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact
     1981 tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg
     2041 acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc
     2101 tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc
     2161 ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga
     2221 aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca
     2281 agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca
     2341 tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag
     2401 aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc
     2461 tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca
     2521 accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg
     2581 ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct
     2641 ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga
     2701 ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg
     2761 agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa
     2821 agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca
     2881 accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg
     2941 actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg
     3001 atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc
     3061 tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc
     3121 tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat
     3181 acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg
     3241 gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc
     3301 tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc
     3361 agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg
     3421 gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc
     3481 ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga
     3541 acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc
     3601 tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc
     3661 tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact
     3721 acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg
     3781 tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg
     3841 tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa
     3901 agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct
     3961 tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg
     4021 actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga
     4081 tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc
     4141 gcgagatcaa caactaccac cacgcccacg acgcctacct aaacgccgtc gtgggaaccg
     4201 ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt
     4261 acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt
     4321 acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg
     4381 agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata
     4441 agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga
     4501 aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca
     4561 gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca
     4621 gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga
     4681 aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga
     4741 agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca
     4801 tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct
     4861 ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc
     4921 tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac
     4981 agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt
     5041 tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca
     5101 agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga
     5161 ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt
     5221 acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt
     5281 acgagacacg gatcgacctg tctcagctgg gaggtgacag cggcgggagc ggcgggagcg
     5341 gggggagcac taatctgagc gacatcattg agaaggagac tgggaaacag ctggtcattc
     5401 aggagtccat cctgatgctg cctgaggagg tggaggaagt gatcggcaac aagccagagt
     5461 ctgacatcct ggtgcacacc gcctacgacg agtccacaga tgagaatgtg atgctgctga
     5521 cctctgacgc ccccgagtat aagccttggg ccctggtcat ccaggattct aacggcgaga
     5581 ataagatcaa gatgctgagc ggaggatccg gaggatctgg aggcagcacc aacctgtctg
     5641 acatcatcga gaaggagaca ggcaagcagc tggtcatcca ggagagcatc ctgatgctgc
     5701 ccgaagaagt cgaagaagtg atcggaaaca agcctgagag cgatatcctg gtccataccg
     5761 cctacgacga gagtaccgac gaaaatgtga tgctgctgac atccgacgcc ccagagtata
     5821 agccctgggc tctggtcatc caggattcca acggagagaa caaaatcaaa atgctgtctg
     5881 gcggctcaaa aagaaccgcc gacggcagcg aattcgagcc caagaagaag aggaaagtcg
     5941 gaagcggagc tactaacttc agcctgctga agcaggctgg agacgtggag gagaaccctg
     6001 gacctatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     6061 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     6121 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     6181 ccaccctcgt gaccaccctg acctatggag tgcagtgctt cagccgctac cccgaccaca
     6241 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     6301 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     6361 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     6421 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     6481 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     6541 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     6601 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     6661 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     6721 agtctggtgg ttctcccaag aagaaaagga aagtctaacc ggtcatcatc accatcacca
     6781 ttgactagta gaccacctcc cctgcgagct aagctggaca gccaatgacg ggtaagagag
     6841 tgacattttt cactaaccta agacaggagg gccgtcagag ctactgccta atccaaagac
     6901 gggtaaaagt gataaaaatg tatcactcca acctaagaca ggcgcagctt ccgagggatt
     6961 tgagatccag acatgataag atacattgat gagtttggac aaaccaaaac tagaatgcag
     7021 tgaaaaaaat gccttatttg tgaaatttgt gatgctattg ccttatttgt aaccattata
     7081 agctgcaata aacaagtttg atatctataa caagaaaata tatatataat aagttatcac
     7141 gtaagtagaa catgaaataa caatataatt atcgtatgag ttaaatctta aaagtcacgt
     7201 aaaagataat catgcgtcat tttgactcac gcggtcgtta tagttcaaaa tcagtgacac
     7261 ttaccgcatt gacaagcacg cctcacggga gctccaagcg gcgactgaga tgtcctaaat
     7321 gcacagcgac ggattcgcgc tatttagaaa gagagagcaa tatttcaaga atgcatgcgt
     7381 caattttacg cagactatct ttctagggtt aagaattcac tggccgtcgt tttacaacgt
     7441 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     7501 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     7561 ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     7621 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     7681 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     7741 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     7801 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     7861 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     7921 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     7981 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     8041 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     8101 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     8161 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     8221 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     8281 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     8341 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     8401 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     8461 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     8521 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     8581 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     8641 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     8701 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     8761 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     8821 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     8881 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     8941 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     9001 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     9061 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     9121 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     9181 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     9241 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     9301 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     9361 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     9421 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     9481 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     9541 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     9601 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     9661 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     9721 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     9781 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     9841 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     9901 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     9961 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
    10021 ggaaacagct atgaccatga ttacgccaag gtcgacttaa ccctagaaag ataatcatat
    10081 tgtgacgtac gttaaagata atcatgcgta aaattgacgc atgtgtttta tcggtctgta
    10141 tatcgaggtt tatttattaa tttgaataga tattaagttt tattatattt acacttacat
    10201 actaataata aattcaacaa acaatttatt tatgtttatt tatttattaa aaaaaaacaa
    10261 aaactcaaaa tttcttctat aaagtaacaa aacttttagc agtgaaaaaa atgctttatt
    10321 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
    10381 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
    10441 taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcctct ggagatttag
    10501 ccctcccaca cataaccaga gggcagcaat tcacgaatcc caactgccgt cggctgtcca
    10561 tcactgtcct tcactatggc tttgatccca ggatgcagat cgagaagcac ctgtcggcac
    10621 cgtccgcagg ggctcaagat gcccctgttc tcatttccga tcgcgacgat acaagtcagg
    10681 ttgccagctg ccgcagcagc agcagtgccc agcaccacga gttctgcaca aggtccccca
    10741 gtaaaatgat atacattgac accagtgaag atgcggccgt cgctagagag agctgcgctg
    10801 gcgacgctgt agtcttcaga gatggggatg ctgttgattg tagccgttgc tctttcaatg
    10861 agggtggatt cttcttgaga caaaggcttg gccatggggc cggggttctc ctccacgtcg
    10921 ccggcctgct tcagcaggct gaagttggtg gcgccgctgc ccccggggag catgtcaagg
    10981 tcaaaatcgt caagagcgtc agcaggcagc atatcaaggt caaagtcgtc aagggcatcg
    11041 gctgggagca tgtctaagtc aaaatcgtca agggcgtcgg tcggcccgcc gctttcgcac
    11101 tttagctgtt tctccaggcc acatatgatt agttccaggc cgaaaaggaa ggcaggttcg
    11161 gctccctgcc ggtcgaacag ctcaattgct tgtttcagaa gtgggggcat agaatcggtg
    11221 gtaggtgtct ctctttcctc ttttgctact tgatgctcct gttcctccaa tacgcagccc
    11281 agtgtaaagt ggcccacggc ggacagagcg tacagtgcgt tctccaggga gaagccttgc
    11341 tgacacagga acgcgagctg attttccagg gtttcgtact gtttctctgt tgggcgggtg
    11401 ccgagatgca ctttagcccc gtcgcgatgt gagaggagag cacagcggta tgacttggcg
    11461 ttgttccgca gaaagtcttg ccatgactcg ccttccaggg ggcaggagtg ggtatgatgc
    11521 ctgtccagca tctcgattgg cagggcatcg agcagggccc gcttgttctt cacgtgccag
    11581 tacagggtag gctgctcaac tcccagcttt tgagcgagtt tccttgtcgt caggccttcg
    11641 ataccgactc cattgagtaa ttccagagca gagtttatga ctttgctctt gtccagtcta
    11701 gacatcttat cgtcatcgtc tttgtaatcc atggtggcgg atcccgcgtc acgacacctg
    11761 tgttctggcg gcaaacccgt tgcgaaaaag aacgttcacg gcgactactg cacttatata
    11821 cggttctccc ccaccctcgg gaaaaaggcg gagccagtac acgacatcac tttcccagtt
    11881 taccccgcgc caccttctct aggcaccggt tcaattgccg acccctcccc ccaacttctc
    11941 ggggactgtg ggcgatgtgc gctctgccca ctgacgggca ccggagccac tcgagtggaa
    12001 ttatcacctc
//
