pET-28b-T7-bLbCas12a-NLS(nucleoplasmin)-6xHis (RTW645)

LOCUS       pET-28b-T7-bLbCas12a-NLS(nucleoplasmin)-6xHis (RTW645)        8982 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 promoter bacterial expression plasmid for bacterial codon 
            optimized LbCas12a with C-terminal NLS and His-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8982)
  AUTHORS   Kleinstiver BP, Sousa AA, Walton RT, Tak YE, Hsu JY, Clement K, 
            Welch MM, Horng JE, Malagon-Lopez J, Scarfo I, Maus MV, Pinello L, 
            Aryee MJ, Joung JK
  TITLE     Engineered CRISPR-Cas12a variants with increased activities and 
            improved targeting ranges for gene, epigenetic and base editing.
  JOURNAL   Nat Biotechnol. 2019 Feb 11. pii: 10.1038/s41587-018-0011-0. doi: 
            10.1038/s41587-018-0011-0.
  PUBMED    30742127
REFERENCE   2  (bases 1 to 8982)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2019 Feb 11. pii: 10.1038/s41587-018-0011-0. doi: 
            10.1038/s41587-018-0011-0."
COMMENT     Sequence Label: pET-28b-T7-bLbCas12a-NLS(nucleoplasmin)-6xHis
            ...
FEATURES             Location/Qualifiers
     source          1..8982
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    5..144
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(70..89)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     230..252
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(246..437)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1246..2328)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2290..2309
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2329..2406)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2612..2631
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2715..2734
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2715..2733
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2734..2758
                     /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)."
     RBS             2773..2795
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2802..6488
                     /codon_start=1
                     /product="CRISPR-associated protein from Lachnospiraceae 
                     bacterium ND2006 (Zetsche et al., 2015)"
                     /label=LbCpf1
                     /translation="MGSKLEKFTNCYSLSKTLRFKAIPVGKTQENIDNKRLLVEDEKRA
                     EDYKGVKKLLDRYYLSFINDVLHSIKLKNLNNYISLFRKKTRTEKENKELENLEINLRK
                     EIAKAFKGNEGYKSLFKKDIIETILPEFLDDKDEIALVNSFNGFTTAFTGFFDNRENMF
                     SEEAKSTSIAFRCINENLTRYISNMDIFEKVDAIFDKHEVQEIKEKILNSDYDVEDFFE
                     GEFFNFVLTQEGIDVYNAIIGGFVTESGEKIKGLNEYINLYNQKTKQKLPKFKPLYKQV
                     LSDRESLSFYGEGYTSDEEVLEVFRNTLNKNSEIFSSIKKLEKLFKNFDEYSSAGIFVK
                     NGPAISTISKDIFGEWNVIRDKWNAEYDDIHLKKKAVVTEKYEDDRRKSFKKIGSFSLE
                     QLQEYADADLSVVEKLKEIIIQKVDEIYKVYGSSEKLFDADFVLEKSLKKNDAVVAIMK
                     DLLDSVKSFENYIKAFFGEGKETNRDESFYGDFVLAYDILLKVDHIYDAIRNYVTQKPY
                     SKDKFKLYFQNPQFMGGWDKDKETDYRATILRYGSKYYLAIMDKKYAKCLQKIDKDDVN
                     GNYEKINYKLLPGPNKMLPKVFFSKKWMAYYNPSEDIQKIYKNGTFKKGDMFNLNDCHK
                     LIDFFKDSISRYPKWSNAYDFNFSETEKYKDIAGFYREVEEQGYKVSFESASKKEVDKL
                     VEEGKLYMFQIYNKDFSDKSHGTPNLHTMYFKLLFDENNHGQIRLSGGAELFMRRASLK
                     KEELVVHPANSPIANKNPDNPKKTTTLSYDVYKDKRFSEDQYELHIPIAINKCPKNIFK
                     INTEVRVLLKHDDNPYVIGIDRGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKT
                     DYHSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALEDLNS
                     GFKNSRVKVEKQVYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMST
                     QNGFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFAL
                     DYKNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINY
                     QQGDIRALLCEQSDKAFYSSFMALMSLMLQMRNSITGRTDVDFLISPVKNSDGIFYDSR
                     NYEAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQT
                     SVKH"
     CDS             6495..6542
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6561..6578
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6631..6649)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6645..6692
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      6729..7184
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6816..6835)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7026..7047
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7276..8091)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(7276..8091)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     7999..8018
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     8083..8102
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      8213..8801
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8702..8721
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8955..8972
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg
       61 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
      121 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
      181 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
      241 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
      301 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
      361 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
      421 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
      481 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
      541 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
      601 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
      661 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
      721 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
      781 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
      841 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
      901 cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
      961 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     1021 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     1081 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     1141 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     1201 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     1261 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     1321 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     1381 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     1441 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     1501 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     1561 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     1621 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     1681 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     1741 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     1801 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     1861 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     1921 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     1981 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     2041 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     2101 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     2161 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     2221 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     2281 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     2341 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     2401 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     2461 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     2521 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     2581 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     2641 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct
     2701 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
     2761 tctagaaata attttgttta actttaagaa ggagatatac catgggcagc aaactggaaa
     2821 aatttacgaa ttgttatagc ctgtccaaga ccctgcgttt caaagccatc cccgttggca
     2881 aaacccagga gaatattgat aataaacgtc tgctggttga ggatgaaaaa agagcagaag
     2941 actataaggg agtcaaaaaa ctgctggatc ggtactacct gagctttata aatgacgtgc
     3001 tgcatagcat taaactgaaa aatctgaata actatattag tctgttccgc aagaaaaccc
     3061 gaacagagaa agaaaataaa gagctggaaa acctggagat caatctgcgt aaagagatcg
     3121 caaaagcttt taaaggaaat gaaggttata aaagcctgtt caaaaaagac attattgaaa
     3181 ccatcctgcc ggaatttctg gatgataaag acgagatagc gctcgtgaac agcttcaacg
     3241 ggttcacgac cgccttcacg ggctttttcg ataacaggga aaatatgttt tcagaggaag
     3301 ccaaaagcac ctcgatagcg ttccgttgca ttaatgaaaa tttgacaaga tatatcagca
     3361 acatggatat tttcgagaaa gttgatgcga tctttgacaa acatgaagtg caggagatta
     3421 aggaaaaaat tctgaacagc gattatgatg ttgaggattt tttcgagggg gaatttttta
     3481 actttgtact gacacaggaa ggtatagatg tgtataatgc tattatcggc gggttcgtta
     3541 ccgaatccgg cgagaaaatt aagggtctga atgagtacat caatctgtat aaccaaaaga
     3601 ccaaacagaa actgccaaaa ttcaaaccgc tgtacaagca agtcctgagc gatcgggaaa
     3661 gcttgagctt ttacggtgaa ggttatacca gcgacgagga ggtactggag gtctttcgca
     3721 ataccctgaa caagaacagc gaaattttca gctccattaa aaagctggag aaactgttta
     3781 agaattttga cgagtacagc agcgcaggta tttttgtgaa gaacggacct gccataagca
     3841 ccattagcaa ggatattttt ggagagtgga atgttatccg tgataaatgg aacgcggaat
     3901 atgatgacat acacctgaaa aagaaggctg tggtaactga gaaatatgaa gacgatcgcc
     3961 gcaaaagctt taaaaaaatc ggcagcttta gcctggagca gctgcaggaa tatgcggacg
     4021 ccgacctgag cgtggtcgag aaactgaagg aaattattat ccaaaaagtg gatgagattt
     4081 acaaggtata tggtagcagc gaaaaactgt ttgatgcgga cttcgttctg gaaaaaagcc
     4141 tgaaaaaaaa tgatgctgtt gttgcgatca tgaaagacct gctcgatagc gttaagagct
     4201 ttgaaaatta cattaaagca ttctttggcg agggcaaaga aacaaacaga gacgaaagct
     4261 tttatggcga cttcgtcctg gcttatgaca tcctgttgaa ggtagatcat atatatgatg
     4321 caattcgtaa ttacgtaacc caaaagccgt acagcaaaga taagttcaaa ctgtatttcc
     4381 agaacccgca gtttatgggt ggctgggaca aagacaagga gacagactat cgcgccacta
     4441 ttctgcgtta cggcagcaag tactatctcg ccatcatgga caaaaaatat gcaaagtgtc
     4501 tgcagaaaat cgataaagac gacgtgaacg gaaattacga aaagattaat tataagctgc
     4561 tgccagggcc caacaagatg ttaccgaaag tatttttttc caaaaaatgg atggcatact
     4621 ataacccgag cgaggatata cagaagattt acaaaaatgg gaccttcaaa aagggggata
     4681 tgttcaatct gaatgactgc cacaaactga tcgatttttt taaagatagc atcagccgtt
     4741 atcctaaatg gtcaaacgcg tatgatttta atttctccga aacggagaaa tataaagaca
     4801 ttgctggttt ctatcgcgaa gtcgaagaac agggttataa agttagcttt gaatcggcca
     4861 gcaagaaaga ggttgataaa ctggtggagg agggtaagct gtatatgttt cagatttata
     4921 acaaagactt tagcgacaaa agccacggta ctcctaatct gcatacgatg tactttaaac
     4981 tgctgtttga tgagaataac cacggccaaa tccgtctctc cggtggagca gaacttttta
     5041 tgcggcgtgc gagcctaaaa aaggaagaac tggtggtgca tcccgccaac agcccgattg
     5101 ctaacaaaaa tccagataat cctaagaaga ccaccacact gtcgtacgat gtctataagg
     5161 ataaacgttt ctcggaagac cagtatgaat tgcatatacc gatagcaatt aataaatgcc
     5221 caaaaaacat tttcaaaatc aacactgaag ttcgtgtgct gctgaaacat gatgataatc
     5281 cgtatgtgat cggaattgac cgtggggaga gaaatctgct gtatattgta gtcgttgatg
     5341 gcaagggcaa catcgttgag cagtatagcc tgaatgaaat aattaataat tttaacggta
     5401 tacgtattaa aaccgactat catagcctgc tggataaaaa ggagaaagag cgttttgagg
     5461 cacgccaaaa ttggacgagc atcgaaaaca tcaaggaact gaaggcagga tatatcagcc
     5521 aagtagtcca taaaatctgt gaactggtgg agaagtacga cgctgtcatt gccctggaag
     5581 acctcaatag cggctttaaa aacagccggg tgaaggtgga gaaacaggta taccaaaagt
     5641 ttgaaaagat gctcattgat aagctgaact atatggttga taaaaagagc aacccgtgcg
     5701 ccactggcgg tgcactgaaa gggtaccaaa ttaccaataa atttgaaagc tttaaaagca
     5761 tgagcacgca gaatgggttt attttttata taccagcatg gctgacgagc aagattgacc
     5821 ccagcactgg ttttgtcaat ctgctgaaaa ccaaatacac aagcattgcg gatagcaaaa
     5881 aatttatttc gagcttcgac cgtattatgt atgttccgga ggaagatctg tttgaatttg
     5941 ccctggatta taaaaacttc agccgcaccg atgcagatta tatcaaaaaa tggaagctgt
     6001 acagttatgg taatcgtata cgtatcttcc gtaatccgaa gaaaaacaat gtgttcgatt
     6061 gggaagaggt ctgtctgacc agcgcgtata aagaactgtt caacaagtac ggaataaatt
     6121 atcagcaagg tgacattcgc gcactgctgt gtgaacagtc agataaagca ttttatagca
     6181 gctttatggc gctgatgagc ctgatgctcc agatgcgcaa cagcataacc ggtcgcacag
     6241 atgttgactt tctgatcagc cctgtgaaga atagcgacgg catcttctac gattccagga
     6301 actatgaagc acaggaaaac gctattctgc ctaaaaatgc cgatgccaac ggcgcctata
     6361 atattgcacg gaaggttctg tgggcgattg gacagttcaa gaaagcggaa gatgagaagc
     6421 tggataaggt aaaaattgct attagcaata aggaatggct ggagtacgca cagacatcgg
     6481 ttaaacacgg tagtaaaagg ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa
     6541 agggagcggc cgcactcgag caccaccacc accaccactg agatccggct gctaacaaag
     6601 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     6661 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg
     6721 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     6781 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     6841 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     6901 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     6961 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     7021 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     7081 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     7141 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     7201 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     7261 ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     7321 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     7381 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     7441 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     7501 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
     7561 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     7621 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     7681 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     7741 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     7801 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     7861 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     7921 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     7981 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     8041 ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct
     8101 tgtattactg tttatgtaag cagacagttt tattgttcat gaccaaaatc ccttaacgtg
     8161 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     8221 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     8281 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     8341 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     8401 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     8461 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     8521 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     8581 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     8641 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     8701 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     8761 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     8821 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     8881 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     8941 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga ag
//