LOCUS       sequence-350234-       12388 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piggyBac transposon containing PE2 and Blasticidin expression 
            cassettes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12388)
  AUTHORS   Wolff JH, Haldrup J, Thomsen EA, Andersen S, Mikkelsen JG
  TITLE     piggyPrime: High-Efficacy Prime Editing in Human Cells Using 
            piggyBac-Based DNA Transposition.
  JOURNAL   Front Genome Ed. 2021 Nov 12;3:786893. doi: 
            10.3389/fgeed.2021.786893. eCollection 2021.
  PUBMED    34870275
REFERENCE   2  (bases 1 to 12388)
  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: "Front 
            Genome Ed."; date: "2021-11-12"; volume: "3:786893. doi"; pages: " 
            10.3389/fgeed.2021.786893. eCollection 2021"
COMMENT     Sequence Label: pPBT-PE2-PGK-Blast
FEATURES             Location/Qualifiers
     source          1..12388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   209..271
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(401..418)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(401..417)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(410..432)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      559..1014
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(646..665)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     856..877
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1040..1144
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1145..1213,1214..2005)
                     /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(1363..1382)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2176..2764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2665..2684
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2918..2935
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3052..3073
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3088..3093,3094..3111,3112..3118)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3126..3142
                     /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     3131..3153
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3150..3166
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3150..3166
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   3657..3691
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        3972..4351
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4352..4555
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4506..4526
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4597..4616
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4597..4615
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4665..4685
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4686..7199,7200..7202,7203..8786)
                     /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             4686..8786
                     /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             10959..10979
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(11005..11022)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11011..11235
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        11245..11755
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     11674..11692
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             11765..12163
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    12164..12388
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 tctagaacta gtgccccggg ctgcaggaat tcgataaaag ttttgttact ttatagaaga
       61 aattttgagt ttttgttttt tttaataaat aaataaacat aaataaattg tttgttgaat
      121 ttattattag tatgtaagtg taaatataat aaaacttaat atctattcaa attaataaat
      181 aaacctcgat atacagaccg ataaaacaca tgcgtcaatt ttacacatga ttatctttaa
      241 cgtacgtcac aatatgatta tctttctagg gttaatctag ctgcgtgttc tgcagcgtgt
      301 cgagcatctt catctgctcc atcacgctgt aaaacacatt tgcaccgcga gtctgcccgt
      361 cctccacggg ttcaaaaacg tgaatgaacg aggcgcgctc actggccgtc gttttacaac
      421 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
      481 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
      541 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
      601 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
      661 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
      721 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
      781 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
      841 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
      901 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
      961 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     1021 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     1081 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     1141 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     1201 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     1261 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     1321 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     1381 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     1441 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     1501 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     1561 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     1621 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     1681 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     1741 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     1801 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     1861 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     1921 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     1981 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     2041 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     2101 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     2161 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2221 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2281 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     2341 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2401 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2461 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2521 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2581 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2641 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2701 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2761 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2821 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2881 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2941 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     3001 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     3061 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     3121 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     3181 gcgcgcccgc cgggtaactc acggggtatc catgtccatt tctgcggcat ccagccagga
     3241 tacccgtcct cgctgacgta atatcccagc gccgcaccgc tgtcattaat ctgcacaccg
     3301 gcacggcagt tccggctgtc gccggtattg ttcgggttgc tgatgcgctt cgggctgacc
     3361 atccggaact gtgtccggaa aagccgcgac gaactggtat cccaggtggc ctgaacgaac
     3421 agttcaccgt taaaggcgtg catggccaca ccttcccgaa tcatcatggt aaacgtgcgt
     3481 tttcgctcaa cgtcaatgca gcagcagtca tcctcggcaa actctttcca tgccgcttca
     3541 acctcgcggg aaaaggcacg ggcttcttcc tccccgatgc ccagatagcg ccagcttggg
     3601 cgatgactga gccggaaaaa agacccgacg atatgatcct gatgcagcta gattaaccct
     3661 agaaagatag tctgcgtaaa attgacgcat gcattcttga aatattgctc tctctttcta
     3721 aatagcgcga atccgtcgct gtgcatttag gacatctcag tcgccgcttg gagctcccgt
     3781 gaggcgtgct tgtcaatgcg gtaagtgtca ctgattttga actataacga ccgcgtgagt
     3841 caaaatgacg catgattatc ttttacgtga cttttaagat ttaactcata cgataattat
     3901 attgttattt catgttctac ttacgtgata acttattata tatatatttt cttgttatag
     3961 atatcaagct tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     4021 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     4081 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     4141 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     4201 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     4261 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     4321 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     4381 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     4441 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     4501 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     4561 agtgaaccgt cagatccgct agagatccgc ggccgctaat acgactcact atagggagag
     4621 ccgccaccat gaaacggaca gccgacggaa gcgagttcga gtcaccaaag aagaagcgga
     4681 aagtcgacaa gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg
     4741 tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc
     4801 ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg
     4861 aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct
     4921 gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca
     4981 gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg
     5041 gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa
     5101 agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc
     5161 acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg
     5221 acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc
     5281 ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca
     5341 gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa
     5401 acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg
     5461 aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg
     5521 cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca
     5581 tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct
     5641 ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc
     5701 ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg
     5761 ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc
     5821 tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc
     5881 ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc
     5941 acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga
     6001 tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca
     6061 gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg
     6121 aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata
     6181 agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg
     6241 tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc
     6301 tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga
     6361 ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa
     6421 tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa
     6481 ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg
     6541 tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg
     6601 cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg
     6661 gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc
     6721 tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg
     6781 acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc
     6841 tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga
     6901 cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg
     6961 tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga
     7021 gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc
     7081 ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc
     7141 gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacg
     7201 ctatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa
     7261 gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga
     7321 agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc
     7381 tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac
     7441 agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga
     7501 acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt
     7561 ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca
     7621 actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa
     7681 agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga
     7741 agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca
     7801 gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc
     7861 ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt
     7921 ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg
     7981 tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga
     8041 tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc cccaccgtgg
     8101 cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg
     8161 tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg
     8221 actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta
     8281 agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac
     8341 tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca
     8401 gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg
     8461 aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag
     8521 tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata
     8581 agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag
     8641 cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca
     8701 aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga
     8761 tcgacctgtc tcagctggga ggtgactctg gaggatctag cggaggatcc tctggcagcg
     8821 agacaccagg aacaagcgag tcagcaacac cagagagcag tggcggcagc agcggcggca
     8881 gcagcaccct aaatatagaa gatgagtatc ggctacatga gacctcaaaa gagccagatg
     8941 tttctctagg gtccacatgg ctgtctgatt ttcctcaggc ctgggcggaa accgggggca
     9001 tgggactggc agttcgccaa gctcctctga tcatacctct gaaagcaacc tctacccccg
     9061 tgtccataaa acaatacccc atgtcacaag aagccagact ggggatcaag ccccacatac
     9121 agagactgtt ggaccaggga atactggtac cctgccagtc cccctggaac acgcccctgc
     9181 tacccgttaa gaaaccaggg actaatgatt ataggcctgt ccaggatctg agagaagtca
     9241 acaagcgggt ggaagacatc caccccaccg tgcccaaccc ttacaacctc ttgagcgggc
     9301 tcccaccgtc ccaccagtgg tacactgtgc ttgatttaaa ggatgccttt ttctgcctga
     9361 gactccaccc caccagtcag cctctcttcg cctttgagtg gagagatcca gagatgggaa
     9421 tctcaggaca attgacctgg accagactcc cacagggttt caaaaacagt cccaccctgt
     9481 ttaatgaggc actgcacaga gacctagcag acttccggat ccagcaccca gacttgatcc
     9541 tgctacagta cgtggatgac ttactgctgg ccgccacttc tgagctagac tgccaacaag
     9601 gtactcgggc cctgttacaa accctaggga acctcgggta tcgggcctcg gccaagaaag
     9661 cccaaatttg ccagaaacag gtcaagtatc tggggtatct tctaaaagag ggtcagagat
     9721 ggctgactga ggccagaaaa gagactgtga tggggcagcc tactccgaag acccctcgac
     9781 aactaaggga gttcctaggg aaggcaggct tctgtcgcct cttcatccct gggtttgcag
     9841 aaatggcagc ccccctgtac cctctcacca aaccggggac tctgtttaat tggggcccag
     9901 accaacaaaa ggcctatcaa gaaatcaagc aagctcttct aactgcccca gccctggggt
     9961 tgccagattt gactaagccc tttgaactct ttgtcgacga gaagcagggc tacgccaaag
    10021 gtgtcctaac gcaaaaactg ggaccttggc gtcggccggt ggcctacctg tccaaaaagc
    10081 tagacccagt agcagctggg tggccccctt gcctacggat ggtagcagcc attgccgtac
    10141 tgacaaagga tgcaggcaag ctaaccatgg gacagccact agtcattctg gccccccatg
    10201 cagtagaggc actagtcaaa caaccccccg accgctggct ttccaacgcc cggatgactc
    10261 actatcaggc cttgcttttg gacacggacc gggtccagtt cggaccggtg gtagccctga
    10321 acccggctac gctgctccca ctgcctgagg aagggctgca acacaactgc cttgatatcc
    10381 tggccgaagc ccacggaacc cgacccgacc taacggacca gccgctccca gacgccgacc
    10441 acacctggta cacggatgga agcagtctct tacaagaggg acagcgtaag gcgggagctg
    10501 cggtgaccac cgagaccgag gtaatctggg ctaaagccct gccagccggg acatccgctc
    10561 agcgggctga actgatagca ctcacccagg ccctaaagat ggcagaaggt aagaagctaa
    10621 atgtttatac tgatagccgt tatgcttttg ctactgccca tatccatgga gaaatataca
    10681 gaaggcgtgg gtggctcaca tcagaaggca aagagatcaa aaataaagac gagatcttgg
    10741 ccctactaaa agccctcttt ctgcccaaaa gacttagcat aatccattgt ccaggacatc
    10801 aaaagggaca cagcgccgag gctagaggca accggatggc tgaccaagcg gcccgaaagg
    10861 cagccatcac agagactcca gacacctcta ccctcctcat agaaaattca tcaccctctg
    10921 gcggctcaaa aagaaccgcc gacggcagcg aattcgagcc caagaagaag aggaaagtct
    10981 aatgtagatc ccccggggga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
    11041 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
    11101 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
    11161 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
    11221 cggtgggctc tatgggcggc cgctggggtt ggggttgcgc cttttccaag gcagccctgg
    11281 gtttgcgcag ggacgcggct gctctgggcg tggttccggg aaacgcagcg gcgccgaccc
    11341 tgggtctcgc acattcttca cgtccgttcg cagcgtcacc cggatcttcg ccgctaccct
    11401 tgtgggcccc ccggcgacgc ttcctgctcc gcccctaagt cgggaaggtt ccttgcggtt
    11461 cgcggcgtgc cggacgtgac aaacggaagc cgcacgtctc actagtaccc tcgcagacgg
    11521 acagcgccag ggagcaatgg cagcgcgccg accgcgatgg gctgtggcca atagcggctg
    11581 ctcagcaggg cgcgccgaga gcagcggccg ggaaggggcg gtgcgggagg cggggtgtgg
    11641 ggcggtagtg tgggccctgt tcctgcccgc gcggtgttcc gcattctgca agcctccgga
    11701 gcgcacgtcg gcagtcggct ccctcgttga ccgaatcacc gacctctctc cccagcaatt
    11761 caccatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag caacggctac
    11821 aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc tctctagcga
    11881 cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt gtgcagaact
    11941 cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat
    12001 cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg tgcttctcga
    12061 tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga cggcagttgg
    12121 gattcgtgaa ttgctgccct ctggttatgt gtgggagggc taactgtgcc ttctagttgc
    12181 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
    12241 actgtccttt cctaataaaa tgagaaaatt gcatcgcatt gtctgagtag gtgtcattct
    12301 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
    12361 catgctgggg atgcggtggg ctctatgg
//
