LOCUS       sequence-309882-       14103 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Piggybac vector that allows for the generation of PE2-expressing 
            stable cell lines.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14103)
  AUTHORS   Melore S, Chavez A
  TITLE     Chavez lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 14103)
  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            "Unpublished"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: pB-CAG-PE2-NatMX
FEATURES             Location/Qualifiers
     source          1..14103
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..659
                     /label=chicken beta-actin promoter
     intron          660..1676
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1600..1622
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1684..1703
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1821..1841
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1842..4355,4356..4358,4359..5942)
                     /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             1842..5942
                     /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             8115..8135
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(8175..8194)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    8219..8267
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        8499..8710
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      8720..8729
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8726..9298
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     polyA_signal    9317..9438
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9354..9373)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9408..9427
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   10201..10235
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(10726..10742)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10726..10742)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10739..10761)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10750..10766
                     /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(10774..10780,10781..10798,10799..10804))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10819..10840
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10957..10974)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11128..11716)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11208..11227)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11887..12678,12679..12747))
                     /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     12510..12529
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12748..12852)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(12878..13333)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(13015..13036)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     13227..13246
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     13460..13482
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     13474..13491
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     13475..13491
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   13621..13683
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gggcgcgcgc cggggggggs ggggsggggs grggggsggg
      541 gsggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      601 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      661 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      781 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      841 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
      961 gcgggcgcgg cgcggggctt tttgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
     1021 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
     1081 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
     1141 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
     1201 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
     1261 ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
     1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
     1381 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
     1441 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
     1501 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
     1561 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
     1621 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
     1681 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaatct agataccctc
     1741 gtaaaggtct agagctagcg aattcgaatt tgagagccgc caccatgaaa cggacagccg
     1801 acggaagcga gttcgagtca ccaaagaaga agcggaaagt cgacaagaag tacagcatcg
     1861 gcctggacat cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc
     1921 ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga
     1981 tcggagccct gctgttcgac agcggcgaaa cagccgaggc cacccggctg aagagaaccg
     2041 ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca
     2101 acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg
     2161 aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct
     2221 accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca
     2281 aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact
     2341 tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc
     2401 tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg
     2461 ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc
     2521 agctgcccgg cgagaagaag aatggcctgt tcggaaacct gattgccctg agcctgggcc
     2581 tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca
     2641 aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg
     2701 acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag
     2761 tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc
     2821 accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca
     2881 aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacattgac ggcggagcca
     2941 gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg
     3001 aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc ttcgacaacg
     3061 gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag
     3121 atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca
     3181 tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa
     3241 agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgcttccg
     3301 cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc
     3361 tgcccaagca cagcctgctg tacgagtact tcaccgtgta taacgagctg accaaagtga
     3421 aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaggcca
     3481 tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact
     3541 acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca
     3601 acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg
     3661 acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg
     3721 acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga
     3781 tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca
     3841 acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct
     3901 tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca
     3961 tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg
     4021 ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg
     4081 tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc
     4141 agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca
     4201 tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga
     4261 acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac
     4321 tggacatcaa ccggctgtcc gactacgatg tggacgctat cgtgcctcag agctttctga
     4381 aggacgactc catcgacaac aaggtgctga ccagaagcga caagaaccgg ggcaagagcg
     4441 acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg cagctgctga
     4501 acgccaagct gattacccag agaaagttcg acaatctgac caaggccgag agaggcggcc
     4561 tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca
     4621 caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac gagaatgaca
     4681 agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga
     4741 aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct
     4801 acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt
     4861 tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg
     4921 aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga
     4981 ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg
     5041 aaaccgggga gatcgtgtgg gataagggcc gggattttgc caccgtgcgg aaagtgctga
     5101 gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag
     5161 agtctatcct gcccaagagg aacagcgata agctgatcgc cagaaagaag gactgggacc
     5221 ctaagaagta cggcggcttc gacagcccca ccgtggccta ttctgtgctg gtggtggcca
     5281 aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg gggatcacca
     5341 tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca
     5401 aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa
     5461 acggccggaa gagaatgctg gcctctgccg gcgaactgca gaagggaaac gaactggccc
     5521 tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct
     5581 cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaagcac tacctggacg
     5641 agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg
     5701 acaaagtgct gtccgcctac aacaagcacc gggataagcc catcagagag caggccgaga
     5761 atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg
     5821 acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac gccaccctga
     5881 tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggtg
     5941 actctggagg atctagcgga ggatcctctg gcagcgagac accaggaaca agcgagtcag
     6001 caacaccaga gagcagtggc ggcagcagcg gcggcagcag caccctaaat atagaagatg
     6061 agtatcggct acatgagacc tcaaaagagc cagatgtttc tctagggtcc acatggctgt
     6121 ctgattttcc tcaggcctgg gcggaaaccg ggggcatggg actggcagtt cgccaagctc
     6181 ctctgatcat acctctgaaa gcaacctcta cccccgtgtc cataaaacaa taccccatgt
     6241 cacaagaagc cagactgggg atcaagcccc acatacagag actgttggac cagggaatac
     6301 tggtaccctg ccagtccccc tggaacacgc ccctgctacc cgttaagaaa ccagggacta
     6361 atgattatag gcctgtccag gatctgagag aagtcaacaa gcgggtggaa gacatccacc
     6421 ccaccgtgcc caacccttac aacctcttga gcgggctccc accgtcccac cagtggtaca
     6481 ctgtgcttga tttaaaggat gcctttttct gcctgagact ccaccccacc agtcagcctc
     6541 tcttcgcctt tgagtggaga gatccagaga tgggaatctc aggacaattg acctggacca
     6601 gactcccaca gggtttcaaa aacagtccca ccctgtttaa tgaggcactg cacagagacc
     6661 tagcagactt ccggatccag cacccagact tgatcctgct acagtacgtg gatgacttac
     6721 tgctggccgc cacttctgag ctagactgcc aacaaggtac tcgggccctg ttacaaaccc
     6781 tagggaacct cgggtatcgg gcctcggcca agaaagccca aatttgccag aaacaggtca
     6841 agtatctggg gtatcttcta aaagagggtc agagatggct gactgaggcc agaaaagaga
     6901 ctgtgatggg gcagcctact ccgaagaccc ctcgacaact aagggagttc ctagggaagg
     6961 caggcttctg tcgcctcttc atccctgggt ttgcagaaat ggcagccccc ctgtaccctc
     7021 tcaccaaacc ggggactctg tttaattggg gcccagacca acaaaaggcc tatcaagaaa
     7081 tcaagcaagc tcttctaact gccccagccc tggggttgcc agatttgact aagccctttg
     7141 aactctttgt cgacgagaag cagggctacg ccaaaggtgt cctaacgcaa aaactgggac
     7201 cttggcgtcg gccggtggcc tacctgtcca aaaagctaga cccagtagca gctgggtggc
     7261 ccccttgcct acggatggta gcagccattg ccgtactgac aaaggatgca ggcaagctaa
     7321 ccatgggaca gccactagtc attctggccc cccatgcagt agaggcacta gtcaaacaac
     7381 cccccgaccg ctggctttcc aacgcccgga tgactcacta tcaggccttg cttttggaca
     7441 cggaccgggt ccagttcgga ccggtggtag ccctgaaccc ggctacgctg ctcccactgc
     7501 ctgaggaagg gctgcaacac aactgccttg atatcctggc cgaagcccac ggaacccgac
     7561 ccgacctaac ggaccagccg ctcccagacg ccgaccacac ctggtacacg gatggaagca
     7621 gtctcttaca agagggacag cgtaaggcgg gagctgcggt gaccaccgag accgaggtaa
     7681 tctgggctaa agccctgcca gccgggacat ccgctcagcg ggctgaactg atagcactca
     7741 cccaggccct aaagatggca gaaggtaaga agctaaatgt ttatactgat agccgttatg
     7801 cttttgctac tgcccatatc catggagaaa tatacagaag gcgtgggtgg ctcacatcag
     7861 aaggcaaaga gatcaaaaat aaagacgaga tcttggccct actaaaagcc ctctttctgc
     7921 ccaaaagact tagcataatc cattgtccag gacatcaaaa gggacacagc gccgaggcta
     7981 gaggcaaccg gatggctgac caagcggccc gaaaggcagc catcacagag actccagaca
     8041 cctctaccct cctcatagaa aattcatcac cctctggcgg ctcaaaaaga accgccgacg
     8101 gcagcgaatt cgagcccaag aagaagagga aagtctgaac cggttagtaa tgagtttaaa
     8161 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     8221 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     8281 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     8341 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     8401 agccaatgtc ggggcggcag gccctgccat agcagatctg cgaaatcgga tccgcggccg
     8461 cactgattct agctgcagag gctccggtgc ccgtcagtgg gcagagcgca catcgcccac
     8521 agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg
     8581 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg
     8641 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc
     8701 cagaacacag gatcctcgag ccaccatggg taccactctt gacgacacgg cttaccggta
     8761 ccgcaccagt gtcccggggg acgccgaggc catcgaggca ctggatgggt ccttcaccac
     8821 cgacaccgtc ttccgcgtca ccgccaccgg ggacggcttc accctgcggg aggtgccggt
     8881 ggacccgccc ctgaccaagg tgttccccga cgacgaatcg gacgacgaat cggacgacgg
     8941 ggaggacggc gacccggact cccggacgtt cgtcgcgtac ggggacgacg gcgacctggc
     9001 gggcttcgtg gttgtctcgt actccggctg gaaccgccgg ctgaccgtcg aggacatcga
     9061 ggtcgccccg gagcaccggg ggcacggggt cgggcgcgcg ttgatggggc tcgcgacgga
     9121 gttcgcccgc gagcggggcg ccgggcacct ctggctggag gtcaccaacg tcaacgcacc
     9181 ggcgatccac gcgtaccggc ggatggggtt caccctctgc ggcctggaca ccgccctgta
     9241 cgacggcacc gcctcggacg gcgagcaggc gctctacatg agcatgccct gcccctaaat
     9301 ctagtgtcga caagttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     9361 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     9421 actcatcaat gtatcttatc atgtctggaa ttgactcaaa tgatgtcaat tagtctatca
     9481 gaagctcatc tggtctccct tccgggggac aagacatccc tgtttaatat ttaaacagca
     9541 gtgttcccaa actgggttct tatatccctt gctctggtca accaggttgc agggtttcct
     9601 gtcctcacag gaacgaagtc cctaaagaaa cagtggcagc caggtttagc cccggaattg
     9661 actggattcc ttttttaggg cccattggta tggctttttc cccgtatccc cccaggtgtc
     9721 tgcaggctca aagagcagcg agaagcgttc agaggaaagc gatcccgtgc caccttcccc
     9781 gtgcccgggc tgtccccgca cgctgccggc tcggggatgc ggggggagcg ccggaccgga
     9841 gcggagcccc gggcggctcg ctgctgcccc ctagcggggg agggacgtaa ttacatccct
     9901 gggggctttg ggggggggct gtccctgata tctataacaa gaaaatatat atataataag
     9961 ttatcacgta agtagaacat gaaataacaa tataattatc gtatgagtta aatcttaaaa
    10021 gtcacgtaaa agataatcat gcgtcatttt gactcacgcg gtcgttatag ttcaaaatca
    10081 gtgacactta ccgcattgac aagcacgcct cacgggagct ccaagcggcg actgagatgt
    10141 cctaaatgca cagcgacgga ttcgcgctat ttagaaagag agagcaatat ttcaagaatg
    10201 catgcgtcaa ttttacgcag actatctttc tagggttaat ctagctgcat caggatcata
    10261 tcgtcgggtc ttttttccgg ctcagtcatc gcccaagctg gcgctatctg ggcatcgggg
    10321 aggaagaagc ccgtgccttt tcccgcgagg ttgaagcggc atggaaagag tttgccgagg
    10381 atgactgctg ctgcattgac gttgagcgaa aacgcacgtt taccatgatg attcgggaag
    10441 gtgtggccat gcacgccttt aacggtgaac tgttcgttca ggccacctgg gataccagtt
    10501 cgtcgcggct tttccggaca cagttccgga tggtcagccc gaagcgcatc agcaacccga
    10561 acaataccgg cgacagccgg aactgccgtg ccggtgtgca gattaatgac agcggtgcgg
    10621 cgctgggata ttacgtcagc gaggacgggt atcctggctg gatgccgcag aaatggacat
    10681 ggataccccg tgagttaccc ggcgggcgcg cttggcgtaa tcatggtcat agctgtttcc
    10741 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
    10801 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
    10861 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
    10921 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
    10981 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
    11041 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
    11101 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
    11161 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
    11221 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
    11281 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
    11341 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
    11401 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
    11461 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
    11521 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
    11581 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
    11641 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
    11701 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
    11761 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
    11821 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
    11881 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
    11941 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
    12001 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
    12061 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
    12121 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
    12181 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
    12241 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
    12301 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
    12361 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
    12421 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
    12481 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
    12541 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
    12601 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
    12661 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
    12721 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
    12781 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
    12841 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
    12901 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
    12961 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
    13021 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
    13081 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
    13141 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
    13201 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
    13261 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
    13321 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
    13381 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
    13441 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcc
    13501 tcgttcattc acgtttttga acccgtggag gacgggcaga ctcgcggtgc aaatgtgttt
    13561 tacagcgtga tggagcagat gaagatgctc gacacgctgc agaacacgca gctagattaa
    13621 ccctagaaag ataatcatat tgtgacgtac gttaaagata atcatgtgta aaattgacgc
    13681 atgtgtttta tcggtctgta tatcgaggtt tatttattaa tttgaataga tattaagttt
    13741 tattatattt acacttacat actaataata aattcaacaa acaatttatt tatgtttatt
    13801 tatttattaa aaaaaacaaa aactcaaaat ttcttctata aagtaacaaa acttttatga
    13861 gggacagccc ccccccaaag cccccaggga tgtaattacg tccctccccc gctagggggc
    13921 agcagcgagc cgcccggggc tccgctccgg tccggcgctc cccccgcatc cccgagccgg
    13981 cagcgtgcgg ggacagcccg ggcacgggga aggtggcacg ggatcgcttt cctctgaacg
    14041 cttctcgctg ctctttgagc ctgcagacac ctggggggat acggggaaaa ggcctccacg
    14101 gcc
//
