LOCUS       sequence-426476-       11816 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piggyBac expression of multiAsCas12a-KRAB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11816)
  AUTHORS   Hsiung CC, Wilson CM, Sambold NA, Dai R, Chen Q, Teyssier N, 
            Misiukiewicz S, Arab A, O'Loughlin T, Cofsky JC, Shi J, Gilbert LA
  TITLE     Engineered CRISPR-Cas12a for higher-order combinatorial chromatin 
            perturbations.
  JOURNAL   Nat Biotechnol. 2024 May 17. doi: 10.1038/s41587-024-02224-0.
  PUBMED    38760567
REFERENCE   2  (bases 1 to 11816)
  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 
            Biotechnol. 2024 May 17. doi: 10.1038/s41587-024-02224-0."
COMMENT     Sequence Label: pCH61 (multiAsCas12a-KRAB piggyBac)
FEATURES             Location/Qualifiers
     source          1..11816
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        147..526
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        528..805
                     /label=chicken beta-actin promoter
     intron          806..1822
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1746..1768
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1830..1849
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1891..1917
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             1927..1953
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             1963..1989
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             1999..2025
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             2035..2061
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             2071..2097
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             2107..6024
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="TQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDH
                     YKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNA
                     IHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDK
                     FTTYFSGFYRNRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFE
                     NVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLN
                     LAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLR
                     NENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGK
                     ITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLK
                     KQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKAR
                     NYATKKPYSVEKFKLNFQMPTLARGWDVNREKNNGAILFVKNGLYYLGIMPKQKGRYKA
                     LSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPL
                     EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL
                     SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH
                     HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD
                     QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFH
                     VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRS
                     LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV
                     VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF
                     TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV
                     KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH
                     RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMA
                     NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK
                     DLKLQNGISNQDWLAYIQELRN"
     CDS             6037..6063
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6073..6093
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6370..6555
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     CDS             6565..6621
                     /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             6622..7329
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMR
                     IKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGV
                     LTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDM
                     ALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVA
                     RYCDLPSKLGHKLN"
     misc_feature    7369..7957
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7422..7442)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7840..7851)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7996..8013)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8002..8226
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   8580..8614
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8701..8720)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8702..8720)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8727..8744)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8727..8743)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8736..8758)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(8885..9340)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9022..9043)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9234..9253
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        9367..9471
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9472..9540,9541..10332)
                     /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(9690..9709)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10503..11091
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10992..11011
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11245..11262
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11379..11400
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11415..11420,11421..11438,11439..11445)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11453..11469
                     /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     11458..11480
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11477..11493
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11477..11493
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11512..11532
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11514..11532
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   11596..11658
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
ORIGIN
        1 aagtaacaaa acttttaaac attctctctt ttacaaaaat aaacttattt tgtactttaa
       61 aaacagtcat gttgtattat aaaataagta attagcttaa cctatacata atagaaacaa
      121 attatactta ttaaccggtc atggtcgaca ttgattattg actagttatt aatagtaatc
      181 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      241 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      301 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      361 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      421 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      481 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
      541 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
      601 tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggc gcgcgccagg
      661 cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa
      721 tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta
      781 taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg
      841 ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt
      901 gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct
      961 tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg
     1021 ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc
     1081 cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag
     1141 tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg
     1201 gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc
     1261 ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg gcccggcttc
     1321 gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg
     1381 gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct cgggggaggg
     1441 gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca gccattgcct
     1501 tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct gtgcggagcc
     1561 gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc
     1621 cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct
     1681 ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg gacggggcag
     1741 ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca
     1801 tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca
     1861 ttttggcaaa gaattgctag cgccaccatg cctgctgcta aacgtgttaa gcttgatggg
     1921 ggtagcccgg cagccaagag agtcaaactc gacggcggct ccccagcggc aaaaagggtg
     1981 aaactagacg ggggtagccc cgccgcgaag cgtgtaaagc tggatggagg atcgcctgcg
     2041 gctaagcgag tcaaattaga cggtgggtca ccggctgcaa agcgcgtcaa attggatgct
     2101 agcggcacac agttcgaggg ctttaccaac ctgtatcagg tgagcaagac actgcggttt
     2161 gagctgatcc cacagggcaa gaccctgaag cacatccagg agcagggctt catcgaggag
     2221 gacaaggccc gcaatgatca ctacaaggag ctgaagccca tcatcgatcg gatctacaag
     2281 acctatgccg accagtgcct gcagctggtg cagctggatt gggagaacct gagcgccgcc
     2341 atcgactcct atagaaagga gaaaaccgag gagacaagga acgccctgat cgaggagcag
     2401 gccacatatc gcaatgccat ccacgactac ttcatcggcc ggacagacaa cctgaccgat
     2461 gccatcaata agagacacgc cgagatctac aagggcctgt tcaaggccga gctgtttaat
     2521 ggcaaggtgc tgaagcagct gggcaccgtg accacaaccg agcacgagaa cgccctgctg
     2581 cggagcttcg acaagtttac aacctacttc tccggctttt atagaaacag gaagaacgtg
     2641 ttcagcgccg aggatatcag cacagccatc ccacaccgca tcgtgcagga caacttcccc
     2701 aagtttaagg agaattgtca catcttcaca cgcctgatca ccgccgtgcc cagcctgcgg
     2761 gagcactttg agaacgtgaa gaaggccatc ggcatcttcg tgagcacctc catcgaggag
     2821 gtgttttcct tcccttttta taaccagctg ctgacacaga cccagatcga cctgtataac
     2881 cagctgctgg gaggaatctc tcgggaggca ggcaccgaga agatcaaggg cctgaacgag
     2941 gtgctgaatc tggccatcca gaagaatgat gagacagccc acatcatcgc ctccctgcca
     3001 cacagattca tccccctgtt taagcagatc ctgtccgata ggaacaccct gtctttcatc
     3061 ctggaggagt ttaagagcga cgaggaagtg atccagtcct tctgcaagta caagacactg
     3121 ctgagaaacg agaacgtgct ggagacagcc gaggccctgt ttaacgagct gaacagcatc
     3181 gacctgacac acatcttcat cagccacaag aagctggaga caatcagcag cgccctgtgc
     3241 gaccactggg atacactgag gaatgccctg tatgagcgga gaatctccga gctgacaggc
     3301 aagatcacca agtctgccaa ggagaaggtg cagcgcagcc tgaagcacga ggatatcaac
     3361 ctgcaggaga tcatctctgc cgcaggcaag gagctgagcg aggccttcaa gcagaaaacc
     3421 agcgagatcc tgtcccacgc acacgccgcc ctggatcagc cactgcctac aaccctgaag
     3481 aagcaggagg agaaggagat cctgaagtct cagctggaca gcctgctggg cctgtaccac
     3541 ctgctggact ggtttgccgt ggatgagtcc aacgaggtgg accccgagtt ctctgcccgg
     3601 ctgaccggca tcaagctgga gatggagcct tctctgagct tctacaacaa ggccagaaat
     3661 tatgccacca agaagcccta ctccgtggag aagttcaagc tgaactttca gatgcctaca
     3721 ctggccagag gctgggacgt gaatagagag aagaacaatg gcgccatcct gtttgtgaag
     3781 aacggcctgt actatctggg catcatgcca aagcagaagg gcaggtataa ggccctgagc
     3841 ttcgagccca cagagaaaac cagcgagggc tttgataaga tgtactatga ctacttccct
     3901 gatgccgcca agatgatccc aaagtgcagc acccagctga aggccgtgac agcccacttt
     3961 cagacccaca caacccccat cctgctgtcc aacaatttca tcgagcctct ggagatcaca
     4021 aaggagatct acgacctgaa caatcctgag aaggagccaa agaagtttca gacagcctac
     4081 gccaagaaaa ccggcgacca gaagggctac agagaggccc tgtgcaagtg gatcgacttc
     4141 acaagggatt ttctgtccaa gtataccaag acaacctcta tcgatctgtc tagcctgcgg
     4201 ccatcctctc agtataagga cctgggcgag tactatgccg agctgaatcc cctgctgtac
     4261 cacatcagct tccagagaat cgccgagaag gagatcatgg atgccgtgga gacaggcaag
     4321 ctgtacctgt tccagatcta taacaaggac tttgccaagg gccaccacgg caagcctaat
     4381 ctgcacacac tgtattggac cggcctgttt tctccagaga acctggccaa gacaagcatc
     4441 aagctgaatg gccaggccga gctgttctac cgccctaagt ccaggatgaa gaggatggca
     4501 caccggctgg gagagaagat gctgaacaag aagctgaagg atcagaaaac cccaatcccc
     4561 gacaccctgt accaggagct gtacgactat gtgaatcaca gactgtccca cgacctgtct
     4621 gatgaggcca gggccctgct gcccaacgtg atcaccaagg aggtgtctca cgagatcatc
     4681 aaggataggc gctttaccag cgacaagttc tttttccacg tgcctatcac actgaactat
     4741 caggccgcca attccccatc taagttcaac cagagggtga atgcctacct gaaggagcac
     4801 cccgagacac ctatcatcgg catcgaccgg ggcgagagaa acctgatcta tatcacagtg
     4861 atcgactcca ccggcaagat cctggagcag cggagcctga acaccatcca gcagtttgat
     4921 taccagaaga agctggacaa cagggagaag gagagggtgg cagcaaggca ggcctggtct
     4981 gtggtgggca caatcaagga tctgaagcag ggctatctga gccaggtcat ccacgagatc
     5041 gtggacctga tgatccacta ccaggccgtg gtggtgctgg agaacctgaa tttcggcttt
     5101 aagagcaaga ggaccggcat cgccgagaag gccgtgtacc agcagttcga gaagatgctg
     5161 atcgataagc tgaattgcct ggtgctgaag gactatccag cagagaaagt gggaggcgtg
     5221 ctgaacccat accagctgac agaccagttc acctcctttg ccaagatggg cacccagtct
     5281 ggcttcctgt tttacgtgcc tgccccatat acatctaaga tcgatcccct gaccggcttc
     5341 gtggacccct tcgtgtggaa aaccatcaag aatcacgaga gccgcaagca cttcctggag
     5401 ggcttcgact ttctgcacta cgacgtgaaa accggcgact tcatcctgca ctttaagatg
     5461 aacagaaatc tgtccttcca gaggggcctg cccggcttta tgcctgcatg ggatatcgtg
     5521 ttcgagaaga acgagacaca gtttgacgcc aagggcaccc ctttcatcgc cggcaagaga
     5581 atcgtgccag tgatcgagaa tcacagattc accggcagat accgggacct gtatcctgcc
     5641 aacgagctga tcgccctgct ggaggagaag ggcatcgtgt tcagggatgg ctccaacatc
     5701 ctgccaaagc tgctggagaa tgacgattct cacgccatcg acacgatggt ggccctgatc
     5761 cgcagcgtgc tgcagatggc caactccaat gccgccacag gcgaggacta tatcaacagc
     5821 cccgtgcgcg atctgaatgg cgtgtgcttc gactcccggt ttcagaaccc agagtggcca
     5881 atggacgccg atgccaatgg cgcctaccac atcgccctga agggccagct gctgctgaat
     5941 cacctgaagg agagcaagga tctgaagctg cagaacggca tctccaatca ggactggctg
     6001 gcctacatcc aggagctgcg caacagcagg gctgactatc cctatgacgt gcccgattat
     6061 gccagcggct cccccaagaa gaagaggaag gtgggatccg gagggccgag ctctggcgca
     6121 cccccaccaa gtggagggtc tcctgccggg tccccaacat ctactgaaga aggcaccagc
     6181 gaatccgcaa cgcccgagtc aggccctggt acctccacag aaccatctga aggtagtgcg
     6241 cctggttccc cagctggaag ccctacttcc accgaagaag gcacgtcaac cgaaccaagt
     6301 gaaggatctg cccctgggac cagcactgaa ccatctgaga tggatgctaa gtcactaact
     6361 gcctggtccc ggacactggt gaccttcaag gatgtatttg tggacttcac cagggaggag
     6421 tggaagctgc tggacactgc tcagcagatc gtgtacagaa atgtgatgct ggagaactat
     6481 aagaacctgg tttccttggg ttatcagctt actaagccag atgtgatcct ccggttggag
     6541 aagggagaag agcccagcgg ccgcgctact aacttcagcc tgctgaagca ggctggagac
     6601 gtggaggaga accctggacc tgtgtctaag ggcgaagagc tgattaagga gaacatgcac
     6661 atgaagctgt acatggaggg caccgtggac aaccatcact tcaagtgcac atccgagggc
     6721 gaaggcaagc cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct
     6781 ctccccttcg ccttcgacat cctggctact agcttcctct acggcagcaa gaccttcatc
     6841 aaccacaccc agggcatccc cgacttcttc aagcagtcct tccctgaggg cttcacatgg
     6901 gagagagtca ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc
     6961 caggacggct gcctcatcta caacgtcaag atcagagggg tgaacttcac atccaacggc
     7021 cctgtgatgc agaagaaaac actcggctgg gaggccttca ccgagacgct gtaccccgct
     7081 gacggcggcc tggaaggcag aaacgacatg gccctgaagc tcgtgggcgg gagccatctg
     7141 atcgcaaacg ccaagaccac atatagatcc aagaaacccg ctaagaacct caagatgcct
     7201 ggcgtctact atgtggacta cagactggaa agaatcaagg aggccaacaa cgagacctac
     7261 gtcgagcagc acgaggtggc agtggccaga tactgcgacc tccctagcaa actggggcac
     7321 aagcttaatt aattaattaa tgatcataca gtgctctgag tacgctataa tcaacctctg
     7381 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     7441 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     7501 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     7561 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     7621 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     7681 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     7741 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     7801 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     7861 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     7921 cagacgagtc ggatctccct ttgggccgcc tccccgcagg atccactaga gcggccgcct
     7981 agagctcgct atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     8041 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     8101 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     8161 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     8221 ctatggcttc tgaggcggaa agaaccagct ggggctcgac tagagcttgc ggaaccctta
     8281 agctttttta ttatatatta gtcacgatat ctataacaag aaaatatata tataataagt
     8341 tatcacgtaa gtagaacatg aaataacaat ataattatcg tatgagttaa atcttaaaag
     8401 tcacgtaaaa gataatcatg cgtcattttg actcacgcgg tcgttatagt tcaaaatcag
     8461 tgacacttac cgcattgaca agcacgcctc acgggagctc caagcggcga ctgagatgtc
     8521 ctaaatgcac agcgacggat tcgcgctatt tagaaagaga gagcaatatt tcaagaatgc
     8581 atgcgtcaat tttacgcaga ctatctttct agggttaaaa aagatttgcg ctttactcga
     8641 cctaaacttt aaacacgtca tagaatcttc gtttgacaaa aaccacattg tgttgggtcg
     8701 ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa
     8761 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     8821 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     8881 tggcaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     8941 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     9001 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     9061 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     9121 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     9181 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     9241 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     9301 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     9361 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     9421 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     9481 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     9541 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     9601 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     9661 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     9721 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     9781 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     9841 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     9901 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     9961 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
    10021 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
    10081 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
    10141 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
    10201 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
    10261 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
    10321 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
    10381 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
    10441 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
    10501 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
    10561 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
    10621 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
    10681 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
    10741 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
    10801 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
    10861 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
    10921 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
    10981 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
    11041 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
    11101 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
    11161 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
    11221 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
    11281 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
    11341 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
    11401 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
    11461 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
    11521 cctcactaaa gggaacctcc cccggtattc acgacagcag gctgaataat aaaaaaatta
    11581 gaaactatta tttaacccta gaaagataat catattgtga cgtacgttaa agataatcat
    11641 gcgtaaaatt gacgcatgtg ttttatcggt ctgtatatcg aggtttattt attaatttga
    11701 atagatatta agttttatta tatttacact tacatactaa taataaattc aacaaacaat
    11761 ttatttatgt ttatttattt attaaaaaaa aacaaaaact caaaatttct tctata
//
