LOCUS       sequence-220510-        9213 bp    DNA     circular SYN 15-APR-2025
DEFINITION  T7 promoter bacterial expression plasmid for human codon optimized 
            AsCas12a-RVR(S542R/K548V/N552R) with C-terminal NLS and His-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9213)
  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 9213)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.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-hAsCas12a-RVR(S542R/K548V/N552R)-NLS(nucleoplasmin)-6xHis
             (AAS1897)
FEATURES             Location/Qualifiers
     source          1..9213
                     /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..6725
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="MGTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARN
                     DHYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYR
                     NAIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSF
                     DKFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREH
                     FENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEV
                     LNLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTL
                     LRNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELT
                     GKITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTT
                     LKKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNK
                     ARNYATKKPYSVEKFKLNFQMPTLARGWDVNVEKNRGAILFVKNGLYYLGIMPKQKGRY
                     KALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIE
                     PLEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSI
                     DLSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAK
                     GHHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKL
                     KDQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFF
                     FHVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQ
                     RSLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQA
                     VVVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTD
                     QFTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHY
                     DVKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIE
                     NHRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQ
                     MRNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKE
                     SKDLKLQNGISNQDWLAYIQELRN"
     CDS             6726..6773
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6792..6809
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6862..6880)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6876..6923
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      6960..7415
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7047..7066)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7257..7278
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7507..8322)
                     /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(7507..8322)
                     /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     8230..8249
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     8314..8333
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      8444..9032
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8933..8952
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9186..9203
                     /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 catgggcaca cagttcgagg
     2821 gctttaccaa cctgtatcag gtgagcaaga cactgcggtt tgagctgatc ccacagggca
     2881 agaccctgaa gcacatccag gagcagggct tcatcgagga ggacaaggcc cgcaatgatc
     2941 actacaagga gctgaagccc atcatcgatc ggatctacaa gacctatgcc gaccagtgcc
     3001 tgcagctggt gcagctggat tgggagaacc tgagcgccgc catcgactcc tatagaaagg
     3061 agaaaaccga ggagacaagg aacgccctga tcgaggagca ggccacatat cgcaatgcca
     3121 tccacgacta cttcatcggc cggacagaca acctgaccga tgccatcaat aagagacacg
     3181 ccgagatcta caagggcctg ttcaaggccg agctgtttaa tggcaaggtg ctgaagcagc
     3241 tgggcaccgt gaccacaacc gagcacgaga acgccctgct gcggagcttc gacaagttta
     3301 caacctactt ctccggcttt tatgagaaca ggaagaacgt gttcagcgcc gaggatatca
     3361 gcacagccat cccacaccgc atcgtgcagg acaacttccc caagtttaag gagaattgtc
     3421 acatcttcac acgcctgatc accgccgtgc ccagcctgcg ggagcacttt gagaacgtga
     3481 agaaggccat cggcatcttc gtgagcacct ccatcgagga ggtgttttcc ttcccttttt
     3541 ataaccagct gctgacacag acccagatcg acctgtataa ccagctgctg ggaggaatct
     3601 ctcgggaggc aggcaccgag aagatcaagg gcctgaacga ggtgctgaat ctggccatcc
     3661 agaagaatga tgagacagcc cacatcatcg cctccctgcc acacagattc atccccctgt
     3721 ttaagcagat cctgtccgat aggaacaccc tgtctttcat cctggaggag tttaagagcg
     3781 acgaggaagt gatccagtcc ttctgcaagt acaagacact gctgagaaac gagaacgtgc
     3841 tggagacagc cgaggccctg tttaacgagc tgaacagcat cgacctgaca cacatcttca
     3901 tcagccacaa gaagctggag acaatcagca gcgccctgtg cgaccactgg gatacactga
     3961 ggaatgccct gtatgagcgg agaatctccg agctgacagg caagatcacc aagtctgcca
     4021 aggagaaggt gcagcgcagc ctgaagcacg aggatatcaa cctgcaggag atcatctctg
     4081 ccgcaggcaa ggagctgagc gaggccttca agcagaaaac cagcgagatc ctgtcccacg
     4141 cacacgccgc cctggatcag ccactgccta caaccctgaa gaagcaggag gagaaggaga
     4201 tcctgaagtc tcagctggac agcctgctgg gcctgtacca cctgctggac tggtttgccg
     4261 tggatgagtc caacgaggtg gaccccgagt tctctgcccg gctgaccggc atcaagctgg
     4321 agatggagcc ttctctgagc ttctacaaca aggccagaaa ttatgccacc aagaagccct
     4381 actccgtgga gaagttcaag ctgaactttc agatgcctac actggccaga ggctgggacg
     4441 tgaatgtgga gaagaacaga ggcgccatcc tgtttgtgaa gaacggcctg tactatctgg
     4501 gcatcatgcc aaagcagaag ggcaggtata aggccctgag cttcgagccc acagagaaaa
     4561 ccagcgaggg ctttgataag atgtactatg actacttccc tgatgccgcc aagatgatcc
     4621 caaagtgcag cacccagctg aaggccgtga cagcccactt tcagacccac acaaccccca
     4681 tcctgctgtc caacaatttc atcgagcctc tggagatcac aaaggagatc tacgacctga
     4741 acaatcctga gaaggagcca aagaagtttc agacagccta cgccaagaaa accggcgacc
     4801 agaagggcta cagagaggcc ctgtgcaagt ggatcgactt cacaagggat tttctgtcca
     4861 agtataccaa gacaacctct atcgatctgt ctagcctgcg gccatcctct cagtataagg
     4921 acctgggcga gtactatgcc gagctgaatc ccctgctgta ccacatcagc ttccagagaa
     4981 tcgccgagaa ggagatcatg gatgccgtgg agacaggcaa gctgtacctg ttccagatct
     5041 ataacaagga ctttgccaag ggccaccacg gcaagcctaa tctgcacaca ctgtattgga
     5101 ccggcctgtt ttctccagag aacctggcca agacaagcat caagctgaat ggccaggccg
     5161 agctgttcta ccgccctaag tccaggatga agaggatggc acaccggctg ggagagaaga
     5221 tgctgaacaa gaagctgaag gatcagaaaa ccccaatccc cgacaccctg taccaggagc
     5281 tgtacgacta tgtgaatcac agactgtccc acgacctgtc tgatgaggcc agggccctgc
     5341 tgcccaacgt gatcaccaag gaggtgtctc acgagatcat caaggatagg cgctttacca
     5401 gcgacaagtt ctttttccac gtgcctatca cactgaacta tcaggccgcc aattccccat
     5461 ctaagttcaa ccagagggtg aatgcctacc tgaaggagca ccccgagaca cctatcatcg
     5521 gcatcgatcg gggcgagaga aacctgatct atatcacagt gatcgactcc accggcaaga
     5581 tcctggagca gcggagcctg aacaccatcc agcagtttga ttaccagaag aagctggaca
     5641 acagggagaa ggagagggtg gcagcaaggc aggcctggtc tgtggtgggc acaatcaagg
     5701 atctgaagca gggctatctg agccaggtca tccacgagat cgtggacctg atgatccact
     5761 accaggccgt ggtggtgctg gagaacctga atttcggctt taagagcaag aggaccggca
     5821 tcgccgagaa ggccgtgtac cagcagttcg agaagatgct gatcgataag ctgaattgcc
     5881 tggtgctgaa ggactatcca gcagagaaag tgggaggcgt gctgaaccca taccagctga
     5941 cagaccagtt cacctccttt gccaagatgg gcacccagtc tggcttcctg ttttacgtgc
     6001 ctgccccata tacatctaag atcgatcccc tgaccggctt cgtggacccc ttcgtgtgga
     6061 aaaccatcaa gaatcacgag agccgcaagc acttcctgga gggcttcgac tttctgcact
     6121 acgacgtgaa aaccggcgac ttcatcctgc actttaagat gaacagaaat ctgtccttcc
     6181 agaggggcct gcccggcttt atgcctgcat gggatatcgt gttcgagaag aacgagacac
     6241 agtttgacgc caagggcacc cctttcatcg ccggcaagag aatcgtgcca gtgatcgaga
     6301 atcacagatt caccggcaga taccgggacc tgtatcctgc caacgagctg atcgccctgc
     6361 tggaggagaa gggcatcgtg ttcagggatg gctccaacat cctgccaaag ctgctggaga
     6421 atgacgattc tcacgccatc gacacgatgg tggccctgat ccgcagcgtg ctgcagatgc
     6481 ggaactccaa tgccgccaca ggcgaggact atatcaacag ccccgtgcgc gatctgaatg
     6541 gcgtgtgctt cgactcccgg tttcagaacc cagagtggcc aatggacgcc gatgccaatg
     6601 gcgcctacca catcgccctg aagggccagc tgctgctgaa tcacctgaag gagagcaagg
     6661 atctgaagct gcagaacggc atctccaatc aggactggct ggcctacatc caggagctgc
     6721 gcaacaaaag gccggcggcc acgaaaaagg ccggccaggc aaaaaagaaa aagggagcgg
     6781 ccgcactcga gcaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg
     6841 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     6901 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga
     6961 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     7021 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     7081 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     7141 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     7201 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     7261 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     7321 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     7381 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     7441 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat
     7501 taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
     7561 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     7621 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     7681 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     7741 gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac
     7801 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     7861 tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg
     7921 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     7981 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca
     8041 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     8101 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     8161 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     8221 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     8281 tcgcggccta gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     8341 gtttatgtaa gcagacagtt ttattgttca tgaccaaaat cccttaacgt gagttttcgt
     8401 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     8461 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     8521 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     8581 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     8641 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     8701 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     8761 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     8821 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     8881 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     8941 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     9001 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     9061 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     9121 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     9181 agcgcagcga gtcagtgagc gaggaagcgg aag
//
