pCAG-enAsCas12a-HF1(E174R/N282A/S542R/K548R)-NLS(nuc)-3xHA (

LOCUS       pCAG-enAsCas12a-HF1(E174R/N282A/S542R/K548R)-NLS(nuc)-3xHA (AAS1815)                8893 bp ds-DNA     circular SYN 07-AUG-2018
DEFINITION  Mammalian expression plasmid for human codon optimized 
            enAsCas12a-HF1 (enhanced high-fidelity AsCas12a) encoding 
            E174R/N282A/S542R/K548R substitutions.
ACCESSION   .
VERSION     .
KEYWORDS    pCAG-enAsCas12a-HF1(E174R/N282A/S542R/K548R)-NLS(nuc)-3xHA (AAS1815)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8893)
  AUTHORS   Keith Joung
  TITLE     Joung lab Cas12a plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8893)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8893
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..662
                     /label=chicken beta-actin promoter
     intron          663..1671
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1595..1617
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1679..1698
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1770..5690
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND
                     HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN
                     AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD
                     KFTTYFSGFYRNRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF
                     ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL
                     ALAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL
                     RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG
                     KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL
                     KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA
                     RNYATKKPYSVEKFKLNFQMPTLARGWDVNREKNNGAILFVKNGLYYLGIMPKQKGRYK
                     ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP
                     LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID
                     LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKG
                     HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK
                     DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFF
                     HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQR
                     SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV
                     VVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ
                     FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD
                     VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN
                     HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM
                     RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES
                     KDLKLQNGISNQDWLAYIQELRN"
     regulatory      5410..5419
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             5691..5738
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5745..5825
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     primer_bind     complement(5915..5934)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_site      5980..6035
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(6034..6053)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6396..6412)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6396..6412)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6409..6431)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6420..6436
                     /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)."
     promoter        complement(6444..6474)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6489..6510
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6569..6764
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6615..6750
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6677..6696
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    6770..6904
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6807..6826)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6861..6880
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6972..6989)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7143..7731)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7223..7242)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7902..8762)
                     /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     8525..8544
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8763..8867)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catgggtcga ggtgagcccc acgttctgct tcactctccc
      421 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
      481 agcgatgggg gcgggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
      541 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgttgcct tcgccccgtg ccccgctccg cgccgcctcg cgccgcccgc
      721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      781 cgggctgtaa ttagcgcttg gtttaatgac ggctcgtttc ttttctgtgg ctgcgtgaaa
      841 gccttaaagg gctccgggag ggccctttgt gcggggggga gcggctcggg gggtgcgtgc
      901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggcccgcgct gcccggcggc tgtgagcgct
      961 gcgggcgcgg cgcggggctt tgtgcgctcc gcgtgtgcgc gaggggagcg cggccggggg
     1021 cggtgccccg cggtgcgggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc
     1081 gtgggggggt gagcaggggg tgtgggcgcg gcggtcgggc tgtaaccccc ccctgcaccc
     1141 ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgtg cggggcgtgg
     1201 cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg
     1261 gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctggcggagc cgaaatctgg gaggcgccgc cgcaccccct
     1441 ctagcgggcg cgggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
     1501 tcgtgcgtcg ccgcgccgcc gtccccttct ccatctccag cctcggggct gccgcagggg
     1561 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     1621 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     1681 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattctgcag tcgacggtac
     1741 cgcgggcccg ggatccaccg gtcgccacca tgacacagtt cgagggcttt accaacctgt
     1801 atcaggtgag caagacactg cggtttgagc tgatcccaca gggcaagacc ctgaagcaca
     1861 tccaggagca gggcttcatc gaggaggaca aggcccgcaa tgatcactac aaggagctga
     1921 agcccatcat cgatcggatc tacaagacct atgccgacca gtgcctgcag ctggtgcagc
     1981 tggattggga gaacctgagc gccgccatcg actcctatag aaaggagaaa accgaggaga
     2041 caaggaacgc cctgatcgag gagcaggcca catatcgcaa tgccatccac gactacttca
     2101 tcggccggac agacaacctg accgatgcca tcaataagag acacgccgag atctacaagg
     2161 gcctgttcaa ggccgagctg tttaatggca aggtgctgaa gcagctgggc accgtgacca
     2221 caaccgagca cgagaacgcc ctgctgcgga gcttcgacaa gtttacaacc tacttctccg
     2281 gcttttatag aaacaggaag aacgtgttca gcgccgagga tatcagcaca gccatcccac
     2341 accgcatcgt gcaggacaac ttccccaagt ttaaggagaa ttgtcacatc ttcacacgcc
     2401 tgatcaccgc cgtgcccagc ctgcgggagc actttgagaa cgtgaagaag gccatcggca
     2461 tcttcgtgag cacctccatc gaggaggtgt tttccttccc tttttataac cagctgctga
     2521 cacagaccca gatcgacctg tataaccagc tgctgggagg aatctctcgg gaggcaggca
     2581 ccgagaagat caagggcctg aacgaggtgc tggccctggc catccagaag aatgatgaga
     2641 cagcccacat catcgcctcc ctgccacaca gattcatccc cctgtttaag cagatcctgt
     2701 ccgataggaa caccctgtct ttcatcctgg aggagtttaa gagcgacgag gaagtgatcc
     2761 agtccttctg caagtacaag acactgctga gaaacgagaa cgtgctggag acagccgagg
     2821 ccctgtttaa cgagctgaac agcatcgacc tgacacacat cttcatcagc cacaagaagc
     2881 tggagacaat cagcagcgcc ctgtgcgacc actgggatac actgaggaat gccctgtatg
     2941 agcggagaat ctccgagctg acaggcaaga tcaccaagtc tgccaaggag aaggtgcagc
     3001 gcagcctgaa gcacgaggat atcaacctgc aggagatcat ctctgccgca ggcaaggagc
     3061 tgagcgaggc cttcaagcag aaaaccagcg agatcctgtc ccacgcacac gccgccctgg
     3121 atcagccact gcctacaacc ctgaagaagc aggaggagaa ggagatcctg aagtctcagc
     3181 tggacagcct gctgggcctg taccacctgc tggactggtt tgccgtggat gagtccaacg
     3241 aggtggaccc cgagttctct gcccggctga ccggcatcaa gctggagatg gagccttctc
     3301 tgagcttcta caacaaggcc agaaattatg ccaccaagaa gccctactcc gtggagaagt
     3361 tcaagctgaa ctttcagatg cctacactgg ccagaggctg ggacgtgaat agagagaaga
     3421 acaatggcgc catcctgttt gtgaagaacg gcctgtacta tctgggcatc atgccaaagc
     3481 agaagggcag gtataaggcc ctgagcttcg agcccacaga gaaaaccagc gagggctttg
     3541 ataagatgta ctatgactac ttccctgatg ccgccaagat gatcccaaag tgcagcaccc
     3601 agctgaaggc cgtgacagcc cactttcaga cccacacaac ccccatcctg ctgtccaaca
     3661 atttcatcga gcctctggag atcacaaagg agatctacga cctgaacaat cctgagaagg
     3721 agccaaagaa gtttcagaca gcctacgcca agaaaaccgg cgaccagaag ggctacagag
     3781 aggccctgtg caagtggatc gacttcacaa gggattttct gtccaagtat accaagacaa
     3841 cctctatcga tctgtctagc ctgcggccat cctctcagta taaggacctg ggcgagtact
     3901 atgccgagct gaatcccctg ctgtaccaca tcagcttcca gagaatcgcc gagaaggaga
     3961 tcatggatgc cgtggagaca ggcaagctgt acctgttcca gatctataac aaggactttg
     4021 ccaagggcca ccacggcaag cctaatctgc acacactgta ttggaccggc ctgttttctc
     4081 cagagaacct ggccaagaca agcatcaagc tgaatggcca ggccgagctg ttctaccgcc
     4141 ctaagtccag gatgaagagg atggcacacc ggctgggaga gaagatgctg aacaagaagc
     4201 tgaaggatca gaaaacccca atccccgaca ccctgtacca ggagctgtac gactatgtga
     4261 atcacagact gtcccacgac ctgtctgatg aggccagggc cctgctgccc aacgtgatca
     4321 ccaaggaggt gtctcacgag atcatcaagg ataggcgctt taccagcgac aagttctttt
     4381 tccacgtgcc tatcacactg aactatcagg ccgccaattc cccatctaag ttcaaccaga
     4441 gggtgaatgc ctacctgaag gagcaccccg agacacctat catcggcatc gatcggggcg
     4501 agagaaacct gatctatatc acagtgatcg actccaccgg caagatcctg gagcagcgga
     4561 gcctgaacac catccagcag tttgattacc agaagaagct ggacaacagg gagaaggaga
     4621 gggtggcagc aaggcaggcc tggtctgtgg tgggcacaat caaggatctg aagcagggct
     4681 atctgagcca ggtcatccac gagatcgtgg acctgatgat ccactaccag gccgtggtgg
     4741 tgctggagaa cctgaatttc ggctttaaga gcaagaggac cggcatcgcc gagaaggccg
     4801 tgtaccagca gttcgagaag atgctgatcg ataagctgaa ttgcctggtg ctgaaggact
     4861 atccagcaga gaaagtggga ggcgtgctga acccatacca gctgacagac cagttcacct
     4921 cctttgccaa gatgggcacc cagtctggct tcctgtttta cgtgcctgcc ccatatacat
     4981 ctaagatcga tcccctgacc ggcttcgtgg accccttcgt gtggaaaacc atcaagaatc
     5041 acgagagccg caagcacttc ctggagggct tcgactttct gcactacgac gtgaaaaccg
     5101 gcgacttcat cctgcacttt aagatgaaca gaaatctgtc cttccagagg ggcctgcccg
     5161 gctttatgcc tgcatgggat atcgtgttcg agaagaacga gacacagttt gacgccaagg
     5221 gcaccccttt catcgccggc aagagaatcg tgccagtgat cgagaatcac agattcaccg
     5281 gcagataccg ggacctgtat cctgccaacg agctgatcgc cctgctggag gagaagggca
     5341 tcgtgttcag ggatggctcc aacatcctgc caaagctgct ggagaatgac gattctcacg
     5401 ccatcgacac catggtggcc ctgatccgca gcgtgctgca gatgcggaac tccaatgccg
     5461 ccacaggcga ggactatatc aacagccccg tgcgcgatct gaatggcgtg tgcttcgact
     5521 cccggtttca gaacccagag tggcccatgg acgccgatgc caatggcgcc taccacatcg
     5581 ccctgaaggg ccagctgctg ctgaatcacc tgaaggagag caaggatctg aagctgcaga
     5641 acggcatctc caatcaggac tggctggcct acatccagga gctgcgcaac aaaaggccgg
     5701 cggccacgaa aaaggccggc caggcaaaaa agaaaaaggg atcctaccca tacgatgttc
     5761 cagattacgc ttatccctac gacgtgcctg attatgcata cccatatgat gtccccgact
     5821 atgcctaaag cggccgcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg
     5881 tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg
     5941 gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca
     6001 ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa
     6061 atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc
     6121 tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg
     6181 ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt
     6241 gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca
     6301 gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat
     6361 ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6421 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6481 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6541 tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc atagtcccgc
     6601 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     6661 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     6721 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta acttgtttat
     6781 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     6841 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     6901 gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     6961 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     7021 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     7081 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     7141 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     7201 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     7261 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     7321 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     7381 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     7441 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     7501 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     7561 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     7621 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     7681 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     7741 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     7801 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     7861 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     7921 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     7981 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     8041 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     8101 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     8161 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     8221 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     8281 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     8341 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     8401 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     8461 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     8521 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     8581 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     8641 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     8701 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     8761 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     8821 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     8881 aaagtgccac ctg
//