LOCUS       sequence-454332-       11859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mU6-driven expression of enAsCas12a gRNAs with 
            EF1a-enAsCas12a-2A-Puro.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11859)
  AUTHORS   El Kassem G, Hillmer J, Boettcher M
  TITLE     Evaluation of Cas13d as a tool for genetic interaction mapping.
  JOURNAL   Nat Commun. 2025 Feb 14;16(1):1631. doi: 10.1038/s41467-025-56747-4.
  PUBMED    39952934
REFERENCE   2  (bases 1 to 11859)
  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: "2025-02-14"; volume: "16(1):1631. doi"; pages: " 
            10.1038/s41467-025-56747-4"
COMMENT     Sequence Label: AiO-Cas12a: All in one enAsCas12a expression a...
FEATURES             Location/Qualifiers
     source          1..11859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(245..256)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             434..667
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    745..866
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      906..1041
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1061..1080)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1062..1080)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1090..1107)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1090..1106)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1099..1121)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1248..1703
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1335..1354)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1545..1566
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1729..1833
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1834..1902,1903..2694)
                     /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(2052..2071)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2865..3453
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3354..3373
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3607..3624
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3741..3762
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3777..3782,3783..3800,3801..3807)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3815..3831
                     /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     3820..3842
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3839..3855
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3839..3855
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3874..3894
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3876..3894
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3922..4148
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4149..4329
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4376..4501
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4994..5227
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5412..5456
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    5637..5754
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     polyA_signal    5813..5934
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6347..6367)
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     protein_bind    6371..6404
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        6714..6925
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             6950..6976
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6977..7003
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7004..7030
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             7031..10948
                     /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"
     regulatory      10668..10677
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             10964..11020
                     /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             11021..11617
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     11514..11534
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    join(11633..11859,1..362)
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11686..11706)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
       61 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
      121 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
      181 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
      241 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
      301 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
      361 gcgtcgactt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
      421 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct
      481 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
      541 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
      601 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
      661 tctagcagta cgtatagtag ttcatgtcat cttattattc agtatttata acttgcaaag
      721 aaatgaatat cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa
      781 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
      841 ttgtccaaac tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc
      901 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
      961 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     1021 gaggaggctt ttttggaggc ctagggacgt acccaattcg ccctatagtg agtcgtatta
     1081 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     1141 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     1201 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     1261 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1321 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1381 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     1441 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     1501 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     1561 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     1621 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     1681 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     1741 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1801 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     1861 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     1921 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     1981 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     2041 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     2101 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     2161 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     2221 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     2281 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     2341 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     2401 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2461 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     2521 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     2581 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     2641 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     2701 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     2761 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     2821 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2881 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2941 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3001 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3061 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3121 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3181 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3241 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3301 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3361 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3421 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3481 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     3541 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     3601 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     3661 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     3721 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     3781 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     3841 ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa
     3901 aagctggagc tgcaagctta atgtagtctt atgcaatact cttgtagtct tgcaacatgg
     3961 taacgatgag ttagcaacat gccttacaag gagagaaaaa gcaccgtgca tgccgattgg
     4021 tggaagtaag gtggtacgat cgtgccttat taggaaggca acagacgggt ctgacatgga
     4081 ttggacgaac cactgaattg ccgcattgca gagatattgt atttaagtgc ctagctcgat
     4141 acataaacgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     4201 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     4261 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     4321 tctctagcag tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc
     4381 tcgacgcagg actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt
     4441 gagtacgcca aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt
     4501 cagtattaag cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg
     4561 aaagaaaaaa tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc
     4621 agttaatcct ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca
     4681 accatccctt cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct
     4741 ctattgtgtg catcaaagga tagagataaa agacaccaag gaagctttag acaagataga
     4801 ggaagagcaa aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg
     4861 gaggaggaga tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa
     4921 ttgaaccatt aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa
     4981 gagcagtggg aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg
     5041 gcgcagcgtc aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc
     5101 agcagaacaa tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct
     5161 ggggcatcaa gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac
     5221 agctcctggg gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga
     5281 atgctagttg gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt
     5341 gggacagaga aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa
     5401 accagcaaga aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga
     5461 attggtttaa cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag
     5521 gcttggtagg tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg
     5581 gatattcacc attatcgttt cagaatcttg agacaaatgg cagtattcat ccacaatttt
     5641 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
     5701 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt
     5761 tattacaggg acagcagaga tccactttgg cgccggcctc gagcagacat gataagatac
     5821 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     5881 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttggattt
     5941 ggatgtgact ggagttcaga cgtgtgctct tccgatcaaa aaaagagacg tacaaaaaag
     6001 agcaagaagc taaaaaagat ttaaaaatta tttttagcgc agttaatgga acaggaacta
     6061 aatttacccc aaagaatcag gatatgaaga tgaaacattt aaaaatgttg taaatccaaa
     6121 tccagaattt gatcctgcat gaaaaatacc gcttgaatat gtttattttg ctcattttta
     6181 ttaaaagttt gcaaaaagga acataaaaat tctaattatt gatactaaag ttattaaaaa
     6241 gaagggtcat agaatataat attgctcatt tacaaccgtc tccccaacaa ggtggttctc
     6301 caagggatac ttatagtctc aaaacacaca attactttac agttagggtg agtttccttt
     6361 tgtgctgttt ataacttcgt ataatgtatg ctatacgaag ttatagaaat ccaagcctat
     6421 catgtaaaat gtagctagta ttaaaaagaa cagattatct gtcttttatc gcacattaag
     6481 cctctatagt tactaggaaa tattatatgc aaattaaccg gggcagggga gtagccgagc
     6541 ttctcccaca agtccgtgcg agggggccgg cgcgggccta gagatggcgc gtcggatcag
     6601 cgctagaaca atggtccata taactccagt tattgaggtt gaagagcatg cccgggctcg
     6661 aggggggccc taggtcttga aaggagtggg aattggctcc ggtgcccgtc agtgggcaga
     6721 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gatccggtgc
     6781 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
     6841 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
     6901 caacgggttt gccgccagaa cacaggaccg gttctagact agagcgatgc ctgctgctaa
     6961 gagagtgaaa ctggatcctg ctgctaagag agtgaaactg gatcctgctg ctaagagagt
     7021 gaaactggat acacagttcg agggctttac caacctgtat caggtgagca agacactgcg
     7081 gtttgagctg atcccacagg gcaagaccct gaagcacatc caggagcagg gcttcatcga
     7141 ggaggacaag gcccgcaatg atcactacaa ggagctgaag cccatcatcg atcggatcta
     7201 caagacctat gccgaccagt gcctgcagct ggtgcagctg gattgggaga acctgagcgc
     7261 cgccatcgac tcctatagaa aggagaaaac cgaggagaca aggaacgccc tgatcgagga
     7321 gcaggccaca tatcgcaatg ccatccacga ctacttcatc ggccggacag acaacctgac
     7381 cgatgccatc aataagagac acgccgagat ctacaagggc ctgttcaagg ccgagctgtt
     7441 taatggcaag gtgctgaagc agctgggcac cgtgaccaca accgagcacg agaacgccct
     7501 gctgcggagc ttcgacaagt ttacaaccta cttctccggc ttttatagaa acaggaagaa
     7561 cgtgttcagc gccgaggata tcagcacagc catcccacac cgcatcgtgc aggacaactt
     7621 ccccaagttt aaggagaatt gtcacatctt cacacgcctg atcaccgccg tgcccagcct
     7681 gcgggagcac tttgagaacg tgaagaaggc catcggcatc ttcgtgagca cctccatcga
     7741 ggaggtgttt tccttccctt tttataacca gctgctgaca cagacccaga tcgacctgta
     7801 taaccagctg ctgggaggaa tctctcggga ggcaggcacc gagaagatca agggcctgaa
     7861 cgaggtgctg aatctggcca tccagaagaa tgatgagaca gcccacatca tcgcctccct
     7921 gccacacaga ttcatccccc tgtttaagca gatcctgtcc gataggaaca ccctgtcttt
     7981 catcctggag gagtttaaga gcgacgagga agtgatccag tccttctgca agtacaagac
     8041 actgctgaga aacgagaacg tgctggagac agccgaggcc ctgtttaacg agctgaacag
     8101 catcgacctg acacacatct tcatcagcca caagaagctg gagacaatca gcagcgccct
     8161 gtgcgaccac tgggatacac tgaggaatgc cctgtatgag cggagaatct ccgagctgac
     8221 aggcaagatc accaagtctg ccaaggagaa ggtgcagcgc agcctgaagc acgaggatat
     8281 caacctgcag gagatcatct ctgccgcagg caaggagctg agcgaggcct tcaagcagaa
     8341 aaccagcgag atcctgtccc acgcacacgc cgccctggat cagccactgc ctacaaccct
     8401 gaagaagcag gaggagaagg agatcctgaa gtctcagctg gacagcctgc tgggcctgta
     8461 ccacctgctg gactggtttg ccgtggatga gtccaacgag gtggaccccg agttctctgc
     8521 ccggctgacc ggcatcaagc tggagatgga gccttctctg agcttctaca acaaggccag
     8581 aaattatgcc accaagaagc cctactccgt ggagaagttc aagctgaact ttcagatgcc
     8641 tacactggcc agaggctggg acgtgaatag agagaagaac aatggcgcca tcctgtttgt
     8701 gaagaacggc ctgtactatc tgggcatcat gccaaagcag aagggcaggt ataaggccct
     8761 gagcttcgag cccacagaga aaaccagcga gggctttgat aagatgtact atgactactt
     8821 ccctgatgcc gccaagatga tcccaaagtg cagcacccag ctgaaggccg tgacagccca
     8881 ctttcagacc cacacaaccc ccatcctgct gtccaacaat ttcatcgagc ctctggagat
     8941 cacaaaggag atctacgacc tgaacaatcc tgagaaggag ccaaagaagt ttcagacagc
     9001 ctacgccaag aaaaccggcg accagaaggg ctacagagag gccctgtgca agtggatcga
     9061 cttcacaagg gattttctgt ccaagtatac caagacaacc tctatcgatc tgtctagcct
     9121 gcggccatcc tctcagtata aggacctggg cgagtactat gccgagctga atcccctgct
     9181 gtaccacatc agcttccaga gaatcgccga gaaggagatc atggatgccg tggagacagg
     9241 caagctgtac ctgttccaga tctataacaa ggactttgcc aagggccacc acggcaagcc
     9301 taatctgcac acactgtatt ggaccggcct gttttctcca gagaacctgg ccaagacaag
     9361 catcaagctg aatggccagg ccgagctgtt ctaccgccct aagtccagga tgaagaggat
     9421 ggcacaccgg ctgggagaga agatgctgaa caagaagctg aaggatcaga aaaccccaat
     9481 ccccgacacc ctgtaccagg agctgtacga ctatgtgaat cacagactgt cccacgacct
     9541 gtctgatgag gccagggccc tgctgcccaa cgtgatcacc aaggaggtgt ctcacgagat
     9601 catcaaggat aggcgcttta ccagcgacaa gttctttttc cacgtgccta tcacactgaa
     9661 ctatcaggcc gccaattccc catctaagtt caaccagagg gtgaatgcct acctgaagga
     9721 gcaccccgag acacctatca tcggcatcga tcggggcgag agaaacctga tctatatcac
     9781 agtgatcgac tccaccggca agatcctgga gcagcggagc ctgaacacca tccagcagtt
     9841 tgattaccag aagaagctgg acaacaggga gaaggagagg gtggcagcaa ggcaggcctg
     9901 gtctgtggtg ggcacaatca aggatctgaa gcagggctat ctgagccagg tcatccacga
     9961 gatcgtggac ctgatgatcc actaccaggc cgtggtggtg ctggagaacc tgaatttcgg
    10021 ctttaagagc aagaggaccg gcatcgccga gaaggccgtg taccagcagt tcgagaagat
    10081 gctgatcgat aagctgaatt gcctggtgct gaaggactat ccagcagaga aagtgggagg
    10141 cgtgctgaac ccataccagc tgacagacca gttcacctcc tttgccaaga tgggcaccca
    10201 gtctggcttc ctgttttacg tgcctgcccc atatacatct aagatcgatc ccctgaccgg
    10261 cttcgtggac cccttcgtgt ggaaaaccat caagaatcac gagagccgca agcacttcct
    10321 ggagggcttc gactttctgc actacgacgt gaaaaccggc gacttcatcc tgcactttaa
    10381 gatgaacaga aatctgtcct tccagagggg cctgcccggc tttatgcctg catgggatat
    10441 cgtgttcgag aagaacgaga cacagtttga cgccaagggc acccctttca tcgccggcaa
    10501 gagaatcgtg ccagtgatcg agaatcacag attcaccggc agataccggg acctgtatcc
    10561 tgccaacgag ctgatcgccc tgctggagga gaagggcatc gtgttcaggg atggctccaa
    10621 catcctgcca aagctgctgg agaatgacga ttctcacgcc atcgacacca tggtggccct
    10681 gatccgcagc gtgctgcaga tgcggaactc caatgccgcc acaggcgagg actatatcaa
    10741 cagccccgtg cgcgatctga atggcgtgtg cttcgactcc cggtttcaga acccagagtg
    10801 gcccatggac gccgatgcca atggcgccta ccacatcgcc ctgaagggcc agctgctgct
    10861 gaatcacctg aaggagagca aggatctgaa gctgcagaac ggcatctcca atcaggactg
    10921 gctggcctac atccaggagc tgcgcaacgc tagcggatcc ggcgcaacaa acttctctct
    10981 gctgaaacaa gccggagatg tcgaagagaa tcctggaccg accgagtaca agcccacggt
    11041 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
    11101 cgactacccc gccacgcgcc acaccgtcga cccggaccgc cacatcgagc gggtcaccga
    11161 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
    11221 cgacggcgcc gccgtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
    11281 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
    11341 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
    11401 cggagtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
    11461 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
    11521 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
    11581 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaacg cgttaagtcg acaatcaacc
    11641 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
    11701 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
    11761 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
    11821 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctga
//
