LOCUS       sequence-244080-       14223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCas9-VPR driven by the bottleneck promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14223)
  AUTHORS   Buchman A, Shriner I, Yang T, Liu J, Antoshechkin I, Marshall JM, 
            Perry MW, Akbari OS
  TITLE     Engineered reproductively isolated species drive reversible 
            population replacement.
  JOURNAL   Nat Commun. 2021 Jun 2;12(1):3281. doi: 10.1038/s41467-021-23531-z.
  PUBMED    34078888
REFERENCE   2  (bases 1 to 14223)
  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: "2021-06-2"; volume: "12(1):3281. doi"; pages: " 
            10.1038/s41467-021-23531-z"
COMMENT     Sequence Label: OA-986C
FEATURES             Location/Qualifiers
     source          1..14223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    270..339
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attB site for the phi-C31 integrase (Groth et al., 
                     2000)"
     regulatory      1256..1265
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1262..1288,1289..1291,1292..3775,3776..3781,3782..3847,
                     3848..3850,3851..5365)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             5378..5398
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5423..5572
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             5597..5617
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6047..6403
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             6422..6991
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     CDS             join(8235..8237,8238..8240,8241..8954)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8280..8301)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8541..8560)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8888..8909
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    9076..9197
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9113..9132)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9167..9186
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   9862..9896
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(10077..10093)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10077..10093)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10090..10112)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10101..10117
                     /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(10125..10131,10132..10149,10150..10155))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10170..10191
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10308..10325)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10479..11067)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10559..10578)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11238..12029,12030..12098))
                     /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     11861..11880
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12099..12203)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     12271..12289
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(12327..12349)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     12449..12468
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12662..12684
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     12676..12693
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     12677..12693
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   13397..13459
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 ataatacata accatttgcg gtttaccggt actttcgttg atagaagcat cctcatcaca
       61 agatgataat aagtatacca tcttagctgg cttcggttta tatgagacga gagtaagggg
      121 tccgtcaaaa caaaacatcg atgttcccac tggcctggag cgactgtttt tcagtacttc
      181 cggtatctcg cgtttgtttg atcgcacggt tcccacaatg gttaattcga gctcgcccgg
      241 ggtcctaggt cgacgatgta ggtcacggtc tcgaagccgc ggtgcgggtg ccagggcgtg
      301 cccttgggct ccccgggcgc gtactccacc tcacccatct ggtccatcat gatgaacggg
      361 tcgaggtggc ggtagttgat cccggcgaac gcgcggcgca ccgggaagcc ctcgccctcg
      421 aaaccgctgg gcgcggtggt cacggtgagc acgggacgtg cgacggcgtc ggcgggtgcg
      481 gatacgcggg gcagcgtcag cgggttctcg acggtcacgg cgggcatgtc gacgcggccg
      541 cattagatga accccatggg atcctgacac ctcatttcgc cttttgcaca gatattttcc
      601 cattccatct gtgtttgatt gcgtatttct cttggaatat atggaaattg cagtagtatg
      661 cttaatgtat aacaccgttt ttatttcgtc cctcatttcc ttctattatt ttgtcggctg
      721 tgaattgcag agtgtcgggt gaatcattaa ttgcattgat tttgatttta acgagccagg
      781 taatgaaacg tcaacagtgt aaagtggcca taaacccctg cggattgctt gtagaaaccc
      841 tgaacatctg tcgcgtttaa tgtttttaag ctatttatga atcgataccc tcgctgatcc
      901 tttttggtgg tccttataaa agtgtgcaga ttagccgtca gtcttttgcc aaaaattggt
      961 gcgcggaaaa cacgcaaaac tcgcgtgttc gggttgcaat taccgggaaa acgctccttt
     1021 gacaaatctt ccttttttac tttcatagct taggtagtgg tctcaggtag tttcccggaa
     1081 ttaagttagg ctgggtatcg cctatcggga acggctacct gaacttttgg caccagctgt
     1141 cgggggtgaa aactggacca ggtagtcttt agaagtgcac ctatataagg tctgcgttcc
     1201 tggcctcaac agcacattcg tacttcaacc gtcaacgaat tcggctgttt aaactgccac
     1261 catggacaag aagtactcca ttgggctcgc tatcggcaca aacagcgtcg gctgggccgt
     1321 cattacggac gagtacaagg tgccgagcaa aaaattcaaa gttctgggca ataccgatcg
     1381 ccacagcata aagaagaacc tcattggcgc cctcctgttc gactccgggg agacggccga
     1441 agccacgcgg ctcaaaagaa cagcacggcg cagatatacc cgcagaaaga atcggatctg
     1501 ctacctgcag gagatcttta gtaatgagat ggctaaggtg gatgactctt tcttccatag
     1561 gctggaggag tcctttttgg tggaggagga taaaaagcac gagcgccacc caatctttgg
     1621 caatatcgtg gacgaggtgg cgtaccatga aaagtaccca accatatatc atctgaggaa
     1681 gaagcttgta gacagtactg ataaggctga cttgcggttg atctatctcg cgctggcgca
     1741 tatgatcaaa tttcggggac acttcctcat cgagggggac ctgaacccag acaacagcga
     1801 tgtcgataaa ctctttatcc aactggttca gacttacaat cagcttttcg aagagaaccc
     1861 gatcaacgca tccggagttg acgccaaagc aatcctgagc gctaggctgt ccaaatcccg
     1921 gcggctcgaa aacctcatcg cacagctccc tggggagaag aagaacggcc tgtttggtaa
     1981 tcttatcgcc ctgtcactcg ggctgacccc caactttaaa tctaacttcg acctggccga
     2041 agatgccaag cttcaactga gcaaagacac ctacgatgat gatctcgaca atctgctggc
     2101 ccagatcggc gaccagtacg cagacctttt tttggcggca aagaacctgt cagacgccat
     2161 tctgctgagt gatattctgc gagtgaacac ggagatcacc aaagctccgc tgagcgctag
     2221 tatgatcaag cgctatgatg agcaccacca agacttgact ttgctgaagg cccttgtcag
     2281 acagcaactg cctgagaagt acaaggaaat tttcttcgat cagtctaaaa atggctacgc
     2341 cggatacatt gacggcggag caagccagga ggaattttac aaatttatta agcccatctt
     2401 ggaaaaaatg gacggcaccg aggagctgct ggtaaagctt aacagagaag atctgttgcg
     2461 caaacagcgc actttcgaca atggaagcat cccccaccag attcacctgg gcgaactgca
     2521 cgctatcctc aggcggcaag aggatttcta cccctttttg aaagataaca gggaaaagat
     2581 tgagaaaatc ctcacatttc ggatacccta ctatgtaggc cccctcgccc ggggaaattc
     2641 cagattcgcg tggatgactc gcaaatcaga agagaccatc actccctgga acttcgagga
     2701 agtcgtggat aagggggcct ctgcccagtc cttcatcgaa aggatgacta actttgataa
     2761 aaatctgcct aacgaaaagg tgcttcctaa acactctctg ctgtacgagt acttcacagt
     2821 ttataacgag ctcaccaagg tcaaatacgt cacagaaggg atgagaaagc cagcattcct
     2881 gtctggagag cagaagaaag ctatcgtgga cctcctcttc aagacgaacc ggaaagttac
     2941 cgtgaaacag ctcaaagaag actatttcaa aaagattgaa tgtttcgact ctgttgaaat
     3001 cagcggagtg gaggatcgct tcaacgcatc cctgggaacg tatcacgatc tcctgaaaat
     3061 cattaaagac aaggacttcc tggacaatga ggagaacgag gacattcttg aggacattgt
     3121 cctcaccctt acgttgtttg aagataggga gatgattgaa gaacgcttga aaacttacgc
     3181 tcatctcttc gacgacaaag tcatgaaaca gctcaagagg cgccgatata caggatgggg
     3241 gcggctgtca agaaaactga tcaatgggat ccgagacaag cagagtggaa agacaatcct
     3301 ggattttctt aagtccgatg gatttgccaa ccggaacttc atgcagttga tccatgatga
     3361 ctctctcacc tttaaggagg acatccagaa agcacaagtt tctggccagg gggacagtct
     3421 tcacgagcac atcgctaatc ttgcaggtag cccagctatc aaaaagggaa tactgcagac
     3481 cgttaaggtc gtggatgaac tcgtcaaagt aatgggaagg cataagcccg agaatatcgt
     3541 tatcgagatg gcccgagaga accaaactac ccagaaggga cagaagaaca gtagggaaag
     3601 gatgaagagg attgaagagg gtataaaaga actggggtcc caaatcctta aggaacaccc
     3661 agttgaaaac acccagcttc agaatgagaa gctctacctg tactacctgc agaacggcag
     3721 ggacatgtac gtggatcagg aactggacat caatcggctc tccgactacg acgtggctgc
     3781 tatcgtgccc cagtcttttc tcaaagatga ttctattgat aataaagtgt tgacaagatc
     3841 cgataaagct agagggaaga gtgataacgt cccctcagaa gaagttgtca agaaaatgaa
     3901 aaattattgg cggcagctgc tgaacgccaa actgatcaca caacggaagt tcgataatct
     3961 gactaaggct gaacgaggtg gcctgtctga gttggataaa gccggcttca tcaaaaggca
     4021 gcttgttgag acacgccaga tcaccaagca cgtggcccaa attctcgatt cacgcatgaa
     4081 caccaagtac gatgaaaatg acaaactgat tcgagaggtg aaagttatta ctctgaagtc
     4141 taagctggtc tcagatttca gaaaggactt tcagttttat aaggtgagag agatcaacaa
     4201 ttaccaccat gcgcatgatg cctacctgaa tgcagtggta ggcactgcac ttatcaaaaa
     4261 atatcccaag cttgaatctg aatttgttta cggagactat aaagtgtacg atgttaggaa
     4321 aatgatcgca aagtctgagc aggaaatagg caaggccacc gctaagtact tcttttacag
     4381 caatattatg aattttttca agaccgagat tacactggcc aatggagaga ttcggaagcg
     4441 accacttatc gaaacaaacg gagaaacagg agaaatcgtg tgggacaagg gtagggattt
     4501 cgcgacagtc cggaaggtcc tgtccatgcc gcaggtgaac atcgttaaaa agaccgaagt
     4561 acagaccgga ggcttctcca aggaaagtat cctcccgaaa aggaacagcg acaagctgat
     4621 cgcacgcaaa aaagattggg accccaagaa atacggcgga ttcgattctc ctacagtcgc
     4681 ttacagtgta ctggttgtgg ccaaagtgga gaaagggaag tctaaaaaac tcaaaagcgt
     4741 caaggaactg ctgggcatca caatcatgga gcgatcaagc ttcgaaaaaa accccatcga
     4801 ctttctcgag gcgaaaggat ataaagaggt caaaaaagac ctcatcatta agcttcccaa
     4861 gtactctctc tttgagcttg aaaacggccg gaaacgaatg ctcgctagtg cgggcgagct
     4921 gcagaaaggt aacgagctgg cactgccctc taaatacgtt aatttcttgt atctggccag
     4981 ccactatgaa aagctcaaag ggtctcccga agataatgag cagaagcagc tgttcgtgga
     5041 acaacacaaa cactaccttg atgagatcat cgagcaaata agcgaattct ccaaaagagt
     5101 gatcctcgcc gacgctaacc tcgataaggt gctttctgct tacaataagc acagggataa
     5161 gcccatcagg gagcaggcag aaaacattat ccacttgttt actctgacca acttgggcgc
     5221 gcctgcagcc ttcaagtact tcgacaccac catagacaga aagcggtaca cctctacaaa
     5281 ggaggtcctg gacgccacac tgattcatca gtcaattacg gggctctatg aaacaagaat
     5341 cgacctctct cagctcggtg gagacagcag ggctgacccc aagaagaaga ggaaggtgga
     5401 ggccagcggt tccggacggg ctgacgcatt ggacgatttt gatctggata tgctgggaag
     5461 tgacgccctc gatgattttg accttgacat gcttggttcg gatgcccttg atgactttga
     5521 cctcgacatg ctcggcagtg acgcccttga tgatttcgac ctggacatgc tgattaactc
     5581 tagaagttcc ggatctccga aaaagaaacg caaagttggt agccagtacc tgcccgacac
     5641 cgacgaccgg caccggatcg aggaaaagcg gaagcggacc tacgagacat tcaagagcat
     5701 catgaagaag tcccccttca gcggccccac cgaccctaga cctccaccta gaagaatcgc
     5761 cgtgcccagc agatccagcg ccagcgtgcc aaaacctgcc ccccagcctt accccttcac
     5821 cagcagcctg agcaccatca actacgacga gttccctacc atggtgttcc ccagcggcca
     5881 gatctctcag gcctctgctc tggctccagc ccctcctcag gtgctgcctc aggctcctgc
     5941 tcctgcacca gctccagcca tggtgtctgc actggctcag gcaccagcac ccgtgcctgt
     6001 gctggctcct ggacctccac aggctgtggc tccaccagcc cctaaaccta cacaggccgg
     6061 cgagggcaca ctgtctgaag ctctgctgca gctgcagttc gacgacgagg atctgggagc
     6121 cctgctggga aacagcaccg atcctgccgt gttcaccgac ctggccagcg tggacaacag
     6181 cgagttccag cagctgctga accagggcat ccctgtggcc cctcacacca ccgagcccat
     6241 gctgatggaa taccccgagg ccatcacccg gctcgtgaca ggcgctcaga ggcctcctga
     6301 tccagctcct gcccctctgg gagcaccagg cctgcctaat ggactgctgt ctggcgacga
     6361 ggacttcagc tctatcgccg acatggactt ctccgcactg ctgggtagcg gatcgggatc
     6421 tcgggattcc agggaaggga tgtttttgcc gaagcctgag gccggctccg ctattagtga
     6481 cgtgtttgag ggccgcgagg tgtgccagcc aaaacgaatc cggccatttc atcctccagg
     6541 aagtccatgg gccaaccgcc cactccccgc cagcctcgca ccaacaccaa ccggtccagt
     6601 acatgagcca gtcgggtcac tgaccccggc accagtccct cagccactgg atccagcgcc
     6661 cgcagtgact cccgaggcca gtcacctgtt ggaggatccc gatgaagaga cgagccaggc
     6721 tgtcaaagcc cttcgggaga tggccgatac tgtgattccc cagaaggaag aggctgcaat
     6781 ctgtggccaa atggaccttt cccatccgcc cccaaggggc catctggatg agctgacaac
     6841 cacacttgag tccatgaccg aggatctgaa cctggactca cccctgaccc cggaattgaa
     6901 cgagattctg gataccttcc tgaacgacga gtgcctcttg catgccatgc atatcagcac
     6961 aggactgtcc atcttcgaca catctctgtt ttgattaatt aactagaatg aatcgttttt
     7021 aaaataacaa atcaattgtt ttataatatt cgtacgattc tttgattatg taataaaatg
     7081 tgatcattag gaagattacg aaaaatataa aaaatatgag ttctgtgtgt ataacaaatg
     7141 ctgtaaacgc cacaattgtg tttgttgcaa ataaacccat gattatttga ttaaaattgt
     7201 tgttttcttt gttcatagac aatagtgtgt tttgcctaaa cgtgtactgc ataaactcca
     7261 tgcgagtgta tagcgagcta gtggctaacg cttgccccac caaagtagat tcgtcaaaat
     7321 cctcaatttc atcaccctcc tccaagttta acatttggcc gtcggaatta acttctaaag
     7381 atgccacata atctaataaa tgaaatagag attcaaacgt ggcgtcatcg tccgtttcga
     7441 ccatttccga aaagaactcg ggcataaact ctatgatttc tctggacgtg gtgttgtcga
     7501 aactctcaaa gtacgcagtc aggaacgtgc gcgacatgtc gtcgggaaac tcgcgcggaa
     7561 acatgttgtt gtaaccgaac gggtcccata gcgccaaaac caaatctgcc agcgtcaata
     7621 gaatgagcac gatgccgaca atggagctgg cttggatagc gattcgagtt aacggccggg
     7681 gcgcgcccat actcggtggc ctccccacca ccaacttttt tgcactgcaa aaaaacacgc
     7741 ttttgcacgc gggcccatac atagtacaaa ctctacgttt cgtagactat tttacataaa
     7801 tagtctacac cgttgtatac gctccaaata cactaccaca cattgaacct ttttgcagtg
     7861 caaaaaagta cgtgtcggca gtcacgtagg ccggccttat cgggtcgcgt cctgtcacgt
     7921 acgaatcaca ttatcggacc ggacgagtgt tgtcttatcg tgacaggacg ccagcttcct
     7981 gtgttgctaa ccgcagccgg acgcaactcc ttatcggaac aggacgcgcc tccatatcag
     8041 ccgcgcgtta tctcatgcgc gtgaccggac acgaggcgcc cgtcccgctt atcgcgccta
     8101 taaatacagc ccgcaacgat ctggtaaaca cagttgaaca gcatctgtta cagcgacaca
     8161 acatgagccg gtccaacaac gccaacgcgc ccacgccatc caaccgccgc cgcaacctgt
     8221 ctctggtgct cgagatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     8281 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     8341 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     8401 tgccctggcc caccctcgtg accaccctga cctggggcgt gcagtgcttc agccgctacc
     8461 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     8521 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     8581 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     8641 acatcctggg gcacaagctg gagtacaact acatcagcca caacgtctat atcaccgccg
     8701 acaagcagaa gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggca
     8761 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     8821 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     8881 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     8941 agctgtacaa gtaagcggcc gcgactctag atcataatca gccataccac atttgtagag
     9001 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     9061 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     9121 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     9181 ctcatcaatg tatcttaaag cttggccggc cgatctcgga tctgacaatg ttcagtgcag
     9241 agactcggct acgcctcgtg gactttgaag ttgaccaaca atgtttattc ttacctctaa
     9301 tagtcctctg tggcaaggtc aagattctgt tagaagccaa tgaagaacct ggttgttcaa
     9361 taacattttg ttcgtctaat atttcactac cgcttgacgt tggctgcact tcatgtacct
     9421 catctataaa cgcttcttct gtatcgctct ggacgtcatc ttcacttacg tgatctgata
     9481 tttcactgtc agaatcctca ccaacaagct cgtcatcgct ttgcagaaga gcagagagga
     9541 tatgctcatc gtctaaagaa ctacccattt tattatatat tagtcacgat atctataaca
     9601 agaaaatata tatataataa gttatcacgt aagtagaaca tgaaataaca atataattat
     9661 cgtatgagtt aaatcttaaa agtcacgtaa aagataatca tgcgtcattt tgactcacgc
     9721 ggtcgttata gttcaaaatc agtgacactt accgcattga caagcacgcc tcacgggagc
     9781 tccaagcggc gactgagatg tcctaaatgc acagcgacgg attcgcgcta tttagaaaga
     9841 gagagcaata tttcaagaat gcatgcgtca attttacgca gactatcttt ctagggttaa
     9901 aaaagatttg cgctttactc gacctaaact ttaaacacgt catagaatct tcgtttgaca
     9961 aaaaccacat tgtggccaag ctgtgtgacg cgacgcgcgc taaagaatgg caaaccaagt
    10021 cgcgcgagcg tcgactctag aggatccccg ggtaccgagc tcgaattcgt aatcatgtca
    10081 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
    10141 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
    10201 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
    10261 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
    10321 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
    10381 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
    10441 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
    10501 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
    10561 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
    10621 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
    10681 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
    10741 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
    10801 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
    10861 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
    10921 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
    10981 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
    11041 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
    11101 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
    11161 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
    11221 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
    11281 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
    11341 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
    11401 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
    11461 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
    11521 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
    11581 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
    11641 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
    11701 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
    11761 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
    11821 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
    11881 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
    11941 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
    12001 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
    12061 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
    12121 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
    12181 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
    12241 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
    12301 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
    12361 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
    12421 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
    12481 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
    12541 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
    12601 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
    12661 tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tttgtttaaa aatataacaa
    12721 aattgtgatc ccacaaaatg aagtggggca aaatcaaata attaatagtg tccgtaaact
    12781 tgttggtctt caactttttg aggaacacgt tggacggcaa atccgtgact ataacacaag
    12841 ttgatttaat aattttagcc aacacgtcgg gctgcgtgtt ttttgccgac gcgtctgtgt
    12901 acacgttgat taactggtcg attaaactgt tgaaataatt taatttttgg ttcttcttta
    12961 aatctgtgat gaaatttttt aaaataactt taaattcttc attggtaaaa aatgccacgt
    13021 tttgcaactt gtgagggtct aatatgaggt caaactcagt aggagtttta tccaaaaaag
    13081 aaaacatgat tacgtctgta cacgaacgcg tattaacgca gagtgcaaag tataagaggg
    13141 ttaaaaaata tattttacgc accatatacg catcgggttg atatcgttaa tatggatcaa
    13201 tttgaacagt tgattaacgt gtctctgctc aagtctttga tcaaaacgca aatcgacgaa
    13261 aatgtgtcgg acaatatcaa gtcgatgagc gaaaaactaa aaaggctaga atacgacaat
    13321 ctcacagaca gcgttgagat atacggtatt cacgacagca ggctgaataa taaaaaaatt
    13381 agaaactatt atttaaccct agaaagataa tcatattgtg acgtacgtta aagataatca
    13441 tgcgtaaaat tgacgcatgt gttttatcgg tctgtatatc gaggtttatt tattaatttg
    13501 aatagatatt aagttttatt atatttacac ttacatacta ataataaatt caacaaacaa
    13561 tttatttatg tttatttatt tattaaaaaa aaacaaaaac tcaaaatttc ttctataaag
    13621 taacaaaact tttaaacatt ctctctttta caaaaataaa cttattttgt actttaaaaa
    13681 cagtcatgtt gtattataaa ataagtaatt agcttaactt atacataata gaaacaaatt
    13741 atacttatta gtcagtcaga aacaactttg gcacatatca atattatgct ctcgacaaat
    13801 aacttttttg cattttttgc acgatgcatt tgcctttcgc cttattttag aggggcagta
    13861 agtacagtaa gtacgttttt tcattactgg ctcttcagta ctgtcatctg atgtaccagg
    13921 cacttcattt ggcaaaatat tagagatatt atcgcgcaaa tatctcttca aagtaggagc
    13981 ttctaaacgc ttacgcataa acgatgacgt caggctcatg taaaggtttc tcataaattt
    14041 tttgcgactt tgaacctttt ctcccttgct actgacatta tggctgtata taataaaaga
    14101 atttatgcag gcaatgttta tcattccgta caataatgcc ataggccacc tattcgtctt
    14161 cctactgcag gtcatcacag aacacatttg gtctagcgtg tccactccgc ctttagtttg
    14221 att
//
