LOCUS       sequence-272540-       11961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses prime editing 2 (PE2) enzyme upon doxycycline treatment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11961)
  TITLE     XLone-PE2 for inducible prime editing
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11961)
  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: 
            "Unpublished"
COMMENT     Sequence Label: XLone-PE2
FEATURES             Location/Qualifiers
     source          1..11961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        9..376
                     /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    17..35
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    53..71
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    89..107
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    125..143
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    161..179
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    197..215
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    233..251
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             425..445
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(446..2959,2960..2962,2963..4546)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(H840A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the H840A mutation in the HNH catalytic domain"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             446..4546
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6719..6739
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    6923..7057
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   7333..7367
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7378..7395)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7378..7394)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7387..7409)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7603..7622)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7722..7744
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7782..7800)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7868..7972
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7973..8041,8042..8833)
                     /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(8191..8210)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9004..9592
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9493..9512
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9746..9763
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9880..9901
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9916..9921,9922..9939,9940..9946)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9954..9970
                     /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     9959..9981
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9978..9994
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9978..9994
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   10020..10082
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    10258..10339
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(10456..10854)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(10855..10911)
                     /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(10921..11664)
                     /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(11665..11688)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      11688..11697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(11717..11928)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
ORIGIN
        1 atcacctcga gtttactccc tatcagtgat agagaacgta tgaagagttt actccctatc
       61 agtgatagag aacgtatgca gactttactc cctatcagtg atagagaacg tataaggagt
      121 ttactcccta tcagtgatag agaacgtatg accagtttac tccctatcag tgatagagaa
      181 cgtatctaca gtttactccc tatcagtgat agagaacgta tatccagttt actccctatc
      241 agtgatagag aacgtataag ctttgcttat gtaaaccagg gcgcctataa aagagtgctg
      301 attttttgag taaacttcaa ttccacaaca cttttgtctt ataccaactt tccgtaccac
      361 ttcctaccct cgtaaaggta ccgccaccat gaaacggaca gccgacggaa gcgagttcga
      421 gtcaccaaag aagaagcgga aagtcgacaa gaagtacagc atcggcctgg acatcggcac
      481 caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa
      541 ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt
      601 cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac
      661 cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt
      721 ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca
      781 cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc
      841 caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct
      901 gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga
      961 cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa
     1021 ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc
     1081 tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa
     1141 gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa
     1201 gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga
     1261 cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc
     1321 caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac
     1381 caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac
     1441 cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga
     1501 ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta
     1561 caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct
     1621 gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca
     1681 gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct
     1741 gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg
     1801 ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat
     1861 caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga
     1921 gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct
     1981 gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg
     2041 aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt
     2101 caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga
     2161 gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac
     2221 ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga
     2281 ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga
     2341 ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg
     2401 gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa
     2461 gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt
     2521 catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt
     2581 gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat
     2641 taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg
     2701 gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg
     2761 acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag
     2821 ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct
     2881 gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct
     2941 gtccgactac gatgtggacg ctatcgtgcc tcagagcttt ctgaaggacg actccatcga
     3001 caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga
     3061 agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac
     3121 ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa
     3181 ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca
     3241 gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt
     3301 gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta
     3361 caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt
     3421 gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta
     3481 caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac
     3541 cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc
     3601 caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt
     3661 gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa
     3721 tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa
     3781 gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg
     3841 cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa
     3901 gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag
     3961 cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga
     4021 cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat
     4081 gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt
     4141 gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga
     4201 gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat
     4261 cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc
     4321 ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt
     4381 taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg
     4441 gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac
     4501 cggcctgtac gagacacgga tcgacctgtc tcagctggga ggtgactctg gaggatctag
     4561 cggaggatcc tctggcagcg agacaccagg aacaagcgag tcagcaacac cagagagcag
     4621 tggcggcagc agcggcggca gcagcaccct aaatatagaa gatgagtatc ggctacatga
     4681 gacctcaaaa gagccagatg tttctctagg gtccacatgg ctgtctgatt ttcctcaggc
     4741 ctgggcggaa accgggggca tgggactggc agttcgccaa gctcctctga tcatacctct
     4801 gaaagcaacc tctacccccg tgtccataaa acaatacccc atgtcacaag aagccagact
     4861 ggggatcaag ccccacatac agagactgtt ggaccaggga atactggtac cctgccagtc
     4921 cccctggaac acgcccctgc tacccgttaa gaaaccaggg actaatgatt ataggcctgt
     4981 ccaggatctg agagaagtca acaagcgggt ggaagacatc caccccaccg tgcccaaccc
     5041 ttacaacctc ttgagcgggc tcccaccgtc ccaccagtgg tacactgtgc ttgatttaaa
     5101 ggatgccttt ttctgcctga gactccaccc caccagtcag cctctcttcg cctttgagtg
     5161 gagagatcca gagatgggaa tctcaggaca attgacctgg accagactcc cacagggttt
     5221 caaaaacagt cccaccctgt ttaatgaggc actgcacaga gacctagcag acttccggat
     5281 ccagcaccca gacttgatcc tgctacagta cgtggatgac ttactgctgg ccgccacttc
     5341 tgagctagac tgccaacaag gtactcgggc cctgttacaa accctaggga acctcgggta
     5401 tcgggcctcg gccaagaaag cccaaatttg ccagaaacag gtcaagtatc tggggtatct
     5461 tctaaaagag ggtcagagat ggctgactga ggccagaaaa gagactgtga tggggcagcc
     5521 tactccgaag acccctcgac aactaaggga gttcctaggg aaggcaggct tctgtcgcct
     5581 cttcatccct gggtttgcag aaatggcagc ccccctgtac cctctcacca aaccggggac
     5641 tctgtttaat tggggcccag accaacaaaa ggcctatcaa gaaatcaagc aagctcttct
     5701 aactgcccca gccctggggt tgccagattt gactaagccc tttgaactct ttgtcgacga
     5761 gaagcagggc tacgccaaag gtgtcctaac gcaaaaactg ggaccttggc gtcggccggt
     5821 ggcctacctg tccaaaaagc tagacccagt agcagctggg tggccccctt gcctacggat
     5881 ggtagcagcc attgccgtac tgacaaagga tgcaggcaag ctaaccatgg gacagccact
     5941 agtcattctg gccccccatg cagtagaggc actagtcaaa caaccccccg accgctggct
     6001 ttccaacgcc cggatgactc actatcaggc cttgcttttg gacacggacc gggtccagtt
     6061 cggaccggtg gtagccctga acccggctac gctgctccca ctgcctgagg aagggctgca
     6121 acacaactgc cttgatatcc tggccgaagc ccacggaacc cgacccgacc taacggacca
     6181 gccgctccca gacgccgacc acacctggta cacggatgga agcagtctct tacaagaggg
     6241 acagcgtaag gcgggagctg cggtgaccac cgagaccgag gtaatctggg ctaaagccct
     6301 gccagccggg acatccgctc agcgggctga actgatagca ctcacccagg ccctaaagat
     6361 ggcagaaggt aagaagctaa atgtttatac tgatagccgt tatgcttttg ctactgccca
     6421 tatccatgga gaaatataca gaaggcgtgg gtggctcaca tcagaaggca aagagatcaa
     6481 aaataaagac gagatcttgg ccctactaaa agccctcttt ctgcccaaaa gacttagcat
     6541 aatccattgt ccaggacatc aaaagggaca cagcgccgag gctagaggca accggatggc
     6601 tgaccaagcg gcccgaaagg cagccatcac agagactcca gacacctcta ccctcctcat
     6661 agaaaattca tcaccctctg gcggctcaaa aagaaccgcc gacggcagcg aattcgagcc
     6721 caagaagaag aggaaagtct aaactagtag accacctccc ctgcgagcta agctggacag
     6781 ccaatgacgg gtaagagagt gacatttttc actaacctaa gacaggaggg ccgtcagagc
     6841 tactgcctaa tccaaagacg ggtaaaagtg ataaaaatgt atcactccaa cctaagacag
     6901 gcgcagcttc cgagggattt gagatccaga catgataaga tacattgatg agtttggaca
     6961 aaccaaaact agaatgcagt gaaaaaaatg ccttatttgt gaaatttgtg atgctattgc
     7021 cttatttgta accattataa gctgcaataa acaagtttga tatctataac aagaaaatat
     7081 atatataata agttatcacg taagtagaac atgaaataac aatataatta tcgtatgagt
     7141 taaatcttaa aagtcacgta aaagataatc atgcgtcatt ttgactcacg cggtcgttat
     7201 agttcaaaat cagtgacact taccgcattg acaagcacgc ctcacgggag ctccaagcgg
     7261 cgactgagat gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat
     7321 atttcaagaa tgcatgcgtc aattttacgc agactatctt tctagggtta agaattcact
     7381 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     7441 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     7501 ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac
     7561 gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc
     7621 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     7681 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     7741 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     7801 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     7861 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     7921 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     7981 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     8041 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     8101 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     8161 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     8221 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     8281 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     8341 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     8401 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     8461 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     8521 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     8581 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     8641 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     8701 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     8761 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     8821 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     8881 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     8941 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     9001 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     9061 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     9121 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     9181 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     9241 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     9301 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     9361 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     9421 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     9481 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     9541 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     9601 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     9661 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     9721 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     9781 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     9841 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     9901 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     9961 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagg tcgacttaac
    10021 cctagaaaga taatcatatt gtgacgtacg ttaaagataa tcatgcgtaa aattgacgca
    10081 tgtgttttat cggtctgtat atcgaggttt atttattaat ttgaatagat attaagtttt
    10141 attatattta cacttacata ctaataataa attcaacaaa caatttattt atgtttattt
    10201 atttattaaa aaaaaacaaa aactcaaaat ttcttctata aagtaacaaa acttttagca
    10261 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
    10321 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
    10381 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta
    10441 tgatcctctg gagatttagc cctcccacac ataaccagag ggcagcaatt cacgaatccc
    10501 aactgccgtc ggctgtccat cactgtcctt cactatggct ttgatcccag gatgcagatc
    10561 gagaagcacc tgtcggcacc gtccgcaggg gctcaagatg cccctgttct catttccgat
    10621 cgcgacgata caagtcaggt tgccagctgc cgcagcagca gcagtgccca gcaccacgag
    10681 ttctgcacaa ggtcccccag taaaatgata tacattgaca ccagtgaaga tgcggccgtc
    10741 gctagagaga gctgcgctgg cgacgctgta gtcttcagag atggggatgc tgttgattgt
    10801 agccgttgct ctttcaatga gggtggattc ttcttgagac aaaggcttgg ccatggggcc
    10861 ggggttctcc tccacgtcgc cggcctgctt cagcaggctg aagttggtgg cgccgctgcc
    10921 cccggggagc atgtcaaggt caaaatcgtc aagagcgtca gcaggcagca tatcaaggtc
    10981 aaagtcgtca agggcatcgg ctgggagcat gtctaagtca aaatcgtcaa gggcgtcggt
    11041 cggcccgccg ctttcgcact ttagctgttt ctccaggcca catatgatta gttccaggcc
    11101 gaaaaggaag gcaggttcgg ctccctgccg gtcgaacagc tcaattgctt gtttcagaag
    11161 tgggggcata gaatcggtgg taggtgtctc tctttcctct tttgctactt gatgctcctg
    11221 ttcctccaat acgcagccca gtgtaaagtg gcccacggcg gacagagcgt acagtgcgtt
    11281 ctccagggag aagccttgct gacacaggaa cgcgagctga ttttccaggg tttcgtactg
    11341 tttctctgtt gggcgggtgc cgagatgcac tttagccccg tcgcgatgtg agaggagagc
    11401 acagcggtat gacttggcgt tgttccgcag aaagtcttgc catgactcgc cttccagggg
    11461 gcaggagtgg gtatgatgcc tgtccagcat ctcgattggc agggcatcga gcagggcccg
    11521 cttgttcttc acgtgccagt acagggtagg ctgctcaact cccagctttt gagcgagttt
    11581 ccttgtcgtc aggccttcga taccgactcc attgagtaat tccagagcag agtttatgac
    11641 tttgctcttg tccagtctag acatcttatc gtcatcgtct ttgtaatcca tggtggcgga
    11701 tcccgcgtca cgacacctgt gttctggcgg caaacccgtt gcgaaaaaga acgttcacgg
    11761 cgactactgc acttatatac ggttctcccc caccctcggg aaaaaggcgg agccagtaca
    11821 cgacatcact ttcccagttt accccgcgcc accttctcta ggcaccggtt caattgccga
    11881 cccctccccc caacttctcg gggactgtgg gcgatgtgcg ctctgcccac tgacgggcac
    11941 cggagccact cgagtggaat t
//
