LOCUS       sequence-426493-       11815 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  piggyBac expresion of enAsCas12a-KRAB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11815)
  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 11815)
  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: pNS49 (enAsCas12a-KRAB piggyBac)
FEATURES             Location/Qualifiers
     source          1..11815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        147..526
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        528..804
                     /label=chicken beta-actin promoter
     intron          805..1821
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1745..1767
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1829..1848
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1890..1916
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             1926..1952
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             1962..1988
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             1998..2024
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             2034..2060
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             2070..2096
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             2106..6023
                     /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
                     RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR
                     NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK
                     DLKLQNGISNQDWLAYIQELRN"
     CDS             6036..6062
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6072..6092
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6369..6554
                     /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             6564..6620
                     /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             6621..7328
                     /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    7368..7956
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7421..7441)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7839..7850)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7995..8012)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8001..8225
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   8579..8613
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8700..8719)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8701..8719)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8726..8743)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8726..8742)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8735..8757)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(8884..9339)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9021..9042)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9233..9252
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        9366..9470
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9471..9539,9540..10331)
                     /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(9689..9708)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10502..11090
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10991..11010
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11244..11261
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11378..11399
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11414..11419,11420..11437,11438..11444)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11452..11468
                     /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     11457..11479
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11476..11492
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11476..11492
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11511..11531
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11513..11531
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   11595..11657
                     /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 gggggggggg cgcgccgggg
      661 ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat
      721 cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat
      781 aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc
      841 tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg
      901 agcgggcggg acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt
      961 gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc gggagggccc tttgtgcggg
     1021 gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc
     1081 gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg ggctttgtgc gctccgcagt
     1141 gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt gcgggggggg ctgcgagggg
     1201 aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg
     1261 gtcgggctgc aaccccccct gcacccccct ccccgagttg ctgagcacgg cccggcttcg
     1321 ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg
     1381 caggtggggg tgccgggcgg ggcggggccg cctcgggccg gggagggctc gggggagggg
     1441 cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt
     1501 ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg tgcggagccg
     1561 aaatctggga ggcgccgccg caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc
     1621 ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc
     1681 cctctccagc ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg
     1741 gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat
     1801 gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat
     1861 tttggcaaag aattgctagc gccaccatgc ctgctgctaa acgtgttaag cttgatgggg
     1921 gtagcccggc agccaagaga gtcaaactcg acggcggctc cccagcggca aaaagggtga
     1981 aactagacgg gggtagcccc gccgcgaagc gtgtaaagct ggatggagga tcgcctgcgg
     2041 ctaagcgagt caaattagac ggtgggtcac cggctgcaaa gcgcgtcaaa ttggatgcta
     2101 gcggcacaca gttcgagggc tttaccaacc tgtatcaggt gagcaagaca ctgcggtttg
     2161 agctgatccc acagggcaag accctgaagc acatccagga gcagggcttc atcgaggagg
     2221 acaaggcccg caatgatcac tacaaggagc tgaagcccat catcgatcgg atctacaaga
     2281 cctatgccga ccagtgcctg cagctggtgc agctggattg ggagaacctg agcgccgcca
     2341 tcgactccta tagaaaggag aaaaccgagg agacaaggaa cgccctgatc gaggagcagg
     2401 ccacatatcg caatgccatc cacgactact tcatcggccg gacagacaac ctgaccgatg
     2461 ccatcaataa gagacacgcc gagatctaca agggcctgtt caaggccgag ctgtttaatg
     2521 gcaaggtgct gaagcagctg ggcaccgtga ccacaaccga gcacgagaac gccctgctgc
     2581 ggagcttcga caagtttaca acctacttct ccggctttta tagaaacagg aagaacgtgt
     2641 tcagcgccga ggatatcagc acagccatcc cacaccgcat cgtgcaggac aacttcccca
     2701 agtttaagga gaattgtcac atcttcacac gcctgatcac cgccgtgccc agcctgcggg
     2761 agcactttga gaacgtgaag aaggccatcg gcatcttcgt gagcacctcc atcgaggagg
     2821 tgttttcctt ccctttttat aaccagctgc tgacacagac ccagatcgac ctgtataacc
     2881 agctgctggg aggaatctct cgggaggcag gcaccgagaa gatcaagggc ctgaacgagg
     2941 tgctgaatct ggccatccag aagaatgatg agacagccca catcatcgcc tccctgccac
     3001 acagattcat ccccctgttt aagcagatcc tgtccgatag gaacaccctg tctttcatcc
     3061 tggaggagtt taagagcgac gaggaagtga tccagtcctt ctgcaagtac aagacactgc
     3121 tgagaaacga gaacgtgctg gagacagccg aggccctgtt taacgagctg aacagcatcg
     3181 acctgacaca catcttcatc agccacaaga agctggagac aatcagcagc gccctgtgcg
     3241 accactggga tacactgagg aatgccctgt atgagcggag aatctccgag ctgacaggca
     3301 agatcaccaa gtctgccaag gagaaggtgc agcgcagcct gaagcacgag gatatcaacc
     3361 tgcaggagat catctctgcc gcaggcaagg agctgagcga ggccttcaag cagaaaacca
     3421 gcgagatcct gtcccacgca cacgccgccc tggatcagcc actgcctaca accctgaaga
     3481 agcaggagga gaaggagatc ctgaagtctc agctggacag cctgctgggc ctgtaccacc
     3541 tgctggactg gtttgccgtg gatgagtcca acgaggtgga ccccgagttc tctgcccggc
     3601 tgaccggcat caagctggag atggagcctt ctctgagctt ctacaacaag gccagaaatt
     3661 atgccaccaa gaagccctac tccgtggaga agttcaagct gaactttcag atgcctacac
     3721 tggccagagg ctgggacgtg aatagagaga agaacaatgg cgccatcctg tttgtgaaga
     3781 acggcctgta ctatctgggc atcatgccaa agcagaaggg caggtataag gccctgagct
     3841 tcgagcccac agagaaaacc agcgagggct ttgataagat gtactatgac tacttccctg
     3901 atgccgccaa gatgatccca aagtgcagca cccagctgaa ggccgtgaca gcccactttc
     3961 agacccacac aacccccatc ctgctgtcca acaatttcat cgagcctctg gagatcacaa
     4021 aggagatcta cgacctgaac aatcctgaga aggagccaaa gaagtttcag acagcctacg
     4081 ccaagaaaac cggcgaccag aagggctaca gagaggccct gtgcaagtgg atcgacttca
     4141 caagggattt tctgtccaag tataccaaga caacctctat cgatctgtct agcctgcggc
     4201 catcctctca gtataaggac ctgggcgagt actatgccga gctgaatccc ctgctgtacc
     4261 acatcagctt ccagagaatc gccgagaagg agatcatgga tgccgtggag acaggcaagc
     4321 tgtacctgtt ccagatctat aacaaggact ttgccaaggg ccaccacggc aagcctaatc
     4381 tgcacacact gtattggacc ggcctgtttt ctccagagaa cctggccaag acaagcatca
     4441 agctgaatgg ccaggccgag ctgttctacc gccctaagtc caggatgaag aggatggcac
     4501 accggctggg agagaagatg ctgaacaaga agctgaagga tcagaaaacc ccaatccccg
     4561 acaccctgta ccaggagctg tacgactatg tgaatcacag actgtcccac gacctgtctg
     4621 atgaggccag ggccctgctg cccaacgtga tcaccaagga ggtgtctcac gagatcatca
     4681 aggataggcg ctttaccagc gacaagttct ttttccacgt gcctatcaca ctgaactatc
     4741 aggccgccaa ttccccatct aagttcaacc agagggtgaa tgcctacctg aaggagcacc
     4801 ccgagacacc tatcatcggc atcgaccggg gcgagagaaa cctgatctat atcacagtga
     4861 tcgactccac cggcaagatc ctggagcagc ggagcctgaa caccatccag cagtttgatt
     4921 accagaagaa gctggacaac agggagaagg agagggtggc agcaaggcag gcctggtctg
     4981 tggtgggcac aatcaaggat ctgaagcagg gctatctgag ccaggtcatc cacgagatcg
     5041 tggacctgat gatccactac caggccgtgg tggtgctgga gaacctgaat ttcggcttta
     5101 agagcaagag gaccggcatc gccgagaagg ccgtgtacca gcagttcgag aagatgctga
     5161 tcgataagct gaattgcctg gtgctgaagg actatccagc agagaaagtg ggaggcgtgc
     5221 tgaacccata ccagctgaca gaccagttca cctcctttgc caagatgggc acccagtctg
     5281 gcttcctgtt ttacgtgcct gccccatata catctaagat cgatcccctg accggcttcg
     5341 tggacccctt cgtgtggaaa accatcaaga atcacgagag ccgcaagcac ttcctggagg
     5401 gcttcgactt tctgcactac gacgtgaaaa ccggcgactt catcctgcac tttaagatga
     5461 acagaaatct gtccttccag aggggcctgc ccggctttat gcctgcatgg gatatcgtgt
     5521 tcgagaagaa cgagacacag tttgacgcca agggcacccc tttcatcgcc ggcaagagaa
     5581 tcgtgccagt gatcgagaat cacagattca ccggcagata ccgggacctg tatcctgcca
     5641 acgagctgat cgccctgctg gaggagaagg gcatcgtgtt cagggatggc tccaacatcc
     5701 tgccaaagct gctggagaat gacgattctc acgccatcga cacgatggtg gccctgatcc
     5761 gcagcgtgct gcagatgcgg aactccaatg ccgccacagg cgaggactat atcaacagcc
     5821 ccgtgcgcga tctgaatggc gtgtgcttcg actcccggtt tcagaaccca gagtggccaa
     5881 tggacgccga tgccaatggc gcctaccaca tcgccctgaa gggccagctg ctgctgaatc
     5941 acctgaagga gagcaaggat ctgaagctgc agaacggcat ctccaatcag gactggctgg
     6001 cctacatcca ggagctgcgc aacagcaggg ctgactatcc ctatgacgtg cccgattatg
     6061 ccagcggctc ccccaagaag aagaggaagg tgggatccgg agggccgagc tctggcgcac
     6121 ccccaccaag tggagggtct cctgccgggt ccccaacatc tactgaagaa ggcaccagcg
     6181 aatccgcaac gcccgagtca ggccctggta cctccacaga accatctgaa ggtagtgcgc
     6241 ctggttcccc agctggaagc cctacttcca ccgaagaagg cacgtcaacc gaaccaagtg
     6301 aaggatctgc ccctgggacc agcactgaac catctgagat ggatgctaag tcactaactg
     6361 cctggtcccg gacactggtg accttcaagg atgtatttgt ggacttcacc agggaggagt
     6421 ggaagctgct ggacactgct cagcagatcg tgtacagaaa tgtgatgctg gagaactata
     6481 agaacctggt ttccttgggt tatcagctta ctaagccaga tgtgatcctc cggttggaga
     6541 agggagaaga gcccagcggc cgcgctacta acttcagcct gctgaagcag gctggagacg
     6601 tggaggagaa ccctggacct gtgtctaagg gcgaagagct gattaaggag aacatgcaca
     6661 tgaagctgta catggagggc accgtggaca accatcactt caagtgcaca tccgagggcg
     6721 aaggcaagcc ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc
     6781 tccccttcgc cttcgacatc ctggctacta gcttcctcta cggcagcaag accttcatca
     6841 accacaccca gggcatcccc gacttcttca agcagtcctt ccctgagggc ttcacatggg
     6901 agagagtcac cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc
     6961 aggacggctg cctcatctac aacgtcaaga tcagaggggt gaacttcaca tccaacggcc
     7021 ctgtgatgca gaagaaaaca ctcggctggg aggccttcac cgagacgctg taccccgctg
     7081 acggcggcct ggaaggcaga aacgacatgg ccctgaagct cgtgggcggg agccatctga
     7141 tcgcaaacgc caagaccaca tatagatcca agaaacccgc taagaacctc aagatgcctg
     7201 gcgtctacta tgtggactac agactggaaa gaatcaagga ggccaacaac gagacctacg
     7261 tcgagcagca cgaggtggca gtggccagat actgcgacct ccctagcaaa ctggggcaca
     7321 agcttaatta attaattaat gatcatacag tgctctgagt acgctataat caacctctgg
     7381 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     7441 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     7501 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     7561 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     7621 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     7681 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     7741 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     7801 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     7861 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     7921 agacgagtcg gatctccctt tgggccgcct ccccgcagga tccactagag cggccgccta
     7981 gagctcgcta tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     8041 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     8101 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     8161 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     8221 tatggcttct gaggcggaaa gaaccagctg gggctcgact agagcttgcg gaacccttaa
     8281 gcttttttat tatatattag tcacgatatc tataacaaga aaatatatat ataataagtt
     8341 atcacgtaag tagaacatga aataacaata taattatcgt atgagttaaa tcttaaaagt
     8401 cacgtaaaag ataatcatgc gtcattttga ctcacgcggt cgttatagtt caaaatcagt
     8461 gacacttacc gcattgacaa gcacgcctca cgggagctcc aagcggcgac tgagatgtcc
     8521 taaatgcaca gcgacggatt cgcgctattt agaaagagag agcaatattt caagaatgca
     8581 tgcgtcaatt ttacgcagac tatctttcta gggttaaaaa agatttgcgc tttactcgac
     8641 ctaaacttta aacacgtcat agaatcttcg tttgacaaaa accacattgt gttgggtcgc
     8701 cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     8761 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     8821 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     8881 ggcaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     8941 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     9001 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     9061 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     9121 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     9181 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     9241 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     9301 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     9361 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     9421 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     9481 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     9541 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     9601 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     9661 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     9721 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     9781 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     9841 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     9901 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     9961 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
    10021 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
    10081 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
    10141 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
    10201 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
    10261 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
    10321 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
    10381 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
    10441 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
    10501 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
    10561 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
    10621 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
    10681 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
    10741 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
    10801 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
    10861 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
    10921 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
    10981 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
    11041 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
    11101 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
    11161 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
    11221 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
    11281 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
    11341 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
    11401 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
    11461 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc
    11521 ctcactaaag ggaacctccc ccggtattca cgacagcagg ctgaataata aaaaaattag
    11581 aaactattat ttaaccctag aaagataatc atattgtgac gtacgttaaa gataatcatg
    11641 cgtaaaattg acgcatgtgt tttatcggtc tgtatatcga ggtttattta ttaatttgaa
    11701 tagatattaa gttttattat atttacactt acatactaat aataaattca acaaacaatt
    11761 tatttatgtt tatttattta ttaaaaaaaa acaaaaactc aaaatttctt ctata
//
