pY108 (lenti-AsCpf1)

LOCUS        pY108 (lenti-AsCpf1)              12863 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Lenti virus delivery of AsCpf1 and crRNA guide.
ACCESSION   .
VERSION     .
KEYWORDS    pY108 (lenti-AsCpf1)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12863)
  AUTHORS   Zetsche B, Heidenreich M, Mohanraju P, Fedorova I, Kneppers J, 
            DeGennaro EM, Winblad N, Choudhury SR, Abudayyeh OO, Gootenberg JS, 
            Wu WY, Scott DA, Severinov K, van der Oost J, Zhang F
  TITLE     Multiplex gene editing by CRISPR-Cpf1 using a single crRNA array.
  JOURNAL   Nat Biotechnol. 2017 Jan;35(1):31-34. doi: 10.1038/nbt.3737. Epub 
            2016 Dec 5.
  PUBMED    27918548
REFERENCE   2  (bases 1 to 12863)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..12863
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        complement(2422..2662)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     complement(2472..2491)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(2642..2662)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        2711..2922
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2930..2939
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2942..2962
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2987..6904
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="TQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDH
                     YKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNA
                     IHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDK
                     FTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFE
                     NVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLN
                     LAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLR
                     NENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGK
                     ITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLK
                     KQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKAR
                     NYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYKA
                     LSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPL
                     EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL
                     SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH
                     HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD
                     QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFH
                     VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRS
                     LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV
                     VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF
                     TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV
                     KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH
                     RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR
                     NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK
                     DLKLQNGISNQDWLAYIQELRN"
     regulatory      6624..6633
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             6905..6952
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6959..7039
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             7049..7105
                     /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             7106..7702
                     /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     7599..7619
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7718..8306
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7771..7791)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8189..8200)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8378..8611
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8637..8654)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8643..8867
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8913..9341
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9000..9019)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9210..9231
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9350..9370)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9355..9684
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9535..9670
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9597..9616
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9732..9779
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9798..10172
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    10302..10423
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10339..10358)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10393..10412
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10472..10488)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10472..10488)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10485..10507)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10496..10512
                     /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(10520..10550)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10565..10586
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10703..10720)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10874..11462)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10954..10973)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(11633..12493)
                     /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     12256..12275
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12494..12598)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(12673..12692)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa tttaattaaa aaagagacga tatatcgtct ccatctacaa
     2401 gagtagaaat tacggtgttt cgtcctttcc acaagatata taaagccaag aaatcgaaat
     2461 actttcaagt tacggtaagc atatgatagt ccattttaaa acataatttt aaaactgcaa
     2521 actacccaag aaattattac tttctacgtc acgtattttg tactaatatc tttgtgttta
     2581 cagtcaaatt aattccaatt atctctctaa cagccttgta tcgtatatgc aaatatgaag
     2641 gaatcatggg aaataggccc tcctcgagag gtcttgaaag gagtgggaat tggctccggt
     2701 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     2761 ggcaattgat ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     2821 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     2881 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggaccggtg ccaccatggc
     2941 cccaaagaag aagcggaagg tcggtatcca cggagtccca gcagccacac agttcgaggg
     3001 ctttaccaac ctgtatcagg tgagcaagac actgcggttt gagctgatcc cacagggcaa
     3061 gaccctgaag cacatccagg agcagggctt catcgaggag gacaaggccc gcaatgatca
     3121 ctacaaggag ctgaagccca tcatcgatcg gatctacaag acctatgccg accagtgcct
     3181 gcagctggtg cagctggatt gggagaacct gagcgccgcc atcgactcct atagaaagga
     3241 gaaaaccgag gagacaagga acgccctgat cgaggagcag gccacatatc gcaatgccat
     3301 ccacgactac ttcatcggcc ggacagacaa cctgaccgat gccatcaata agagacacgc
     3361 cgagatctac aagggcctgt tcaaggccga gctgtttaat ggcaaggtgc tgaagcagct
     3421 gggcaccgtg accacaaccg agcacgagaa cgccctgctg cggagcttcg acaagtttac
     3481 aacctacttc tccggctttt atgagaacag gaagaacgtg ttcagcgccg aggatatcag
     3541 cacagccatc ccacaccgca tcgtgcagga caacttcccc aagtttaagg agaattgtca
     3601 catcttcaca cgcctgatca ccgccgtgcc cagcctgcgg gagcactttg agaacgtgaa
     3661 gaaggccatc ggcatcttcg tgagcacctc catcgaggag gtgttttcct tcccttttta
     3721 taaccagctg ctgacacaga cccagatcga cctgtataac cagctgctgg gaggaatctc
     3781 tcgggaggca ggcaccgaga agatcaaggg cctgaacgag gtgctgaatc tggccatcca
     3841 gaagaatgat gagacagccc acatcatcgc ctccctgcca cacagattca tccccctgtt
     3901 taagcagatc ctgtccgata ggaacaccct gtctttcatc ctggaggagt ttaagagcga
     3961 cgaggaagtg atccagtcct tctgcaagta caagacactg ctgagaaacg agaacgtgct
     4021 ggagacagcc gaggccctgt ttaacgagct gaacagcatc gacctgacac acatcttcat
     4081 cagccacaag aagctggaga caatcagcag cgccctgtgc gaccactggg atacactgag
     4141 gaatgccctg tatgagcgga gaatctccga gctgacaggc aagatcacca agtctgccaa
     4201 ggagaaggtg cagcgcagcc tgaagcacga ggatatcaac ctgcaggaga tcatctctgc
     4261 cgcaggcaag gagctgagcg aggccttcaa gcagaaaacc agcgagatcc tgtcccacgc
     4321 acacgccgcc ctggatcagc cactgcctac aaccctgaag aagcaggagg agaaggagat
     4381 cctgaagtct cagctggaca gcctgctggg cctgtaccac ctgctggact ggtttgccgt
     4441 ggatgagtcc aacgaggtgg accccgagtt ctctgcccgg ctgaccggca tcaagctgga
     4501 gatggagcct tctctgagct tctacaacaa ggccagaaat tatgccacca agaagcccta
     4561 ctccgtggag aagttcaagc tgaactttca gatgcctaca ctggcctctg gctgggacgt
     4621 gaataaggag aagaacaatg gcgccatcct gtttgtgaag aacggcctgt actatctggg
     4681 catcatgcca aagcagaagg gcaggtataa ggccctgagc ttcgagccca cagagaaaac
     4741 cagcgagggc tttgataaga tgtactatga ctacttccct gatgccgcca agatgatccc
     4801 aaagtgcagc acccagctga aggccgtgac agcccacttt cagacccaca caacccccat
     4861 cctgctgtcc aacaatttca tcgagcctct ggagatcaca aaggagatct acgacctgaa
     4921 caatcctgag aaggagccaa agaagtttca gacagcctac gccaagaaaa ccggcgacca
     4981 gaagggctac agagaggccc tgtgcaagtg gatcgacttc acaagggatt ttctgtccaa
     5041 gtataccaag acaacctcta tcgatctgtc tagcctgcgg ccatcctctc agtataagga
     5101 cctgggcgag tactatgccg agctgaatcc cctgctgtac cacatcagct tccagagaat
     5161 cgccgagaag gagatcatgg atgccgtgga gacaggcaag ctgtacctgt tccagatcta
     5221 taacaaggac tttgccaagg gccaccacgg caagcctaat ctgcacacac tgtattggac
     5281 cggcctgttt tctccagaga acctggccaa gacaagcatc aagctgaatg gccaggccga
     5341 gctgttctac cgccctaagt ccaggatgaa gaggatggca caccggctgg gagagaagat
     5401 gctgaacaag aagctgaagg atcagaaaac cccaatcccc gacaccctgt accaggagct
     5461 gtacgactat gtgaatcaca gactgtccca cgacctgtct gatgaggcca gggccctgct
     5521 gcccaacgtg atcaccaagg aggtgtctca cgagatcatc aaggataggc gctttaccag
     5581 cgacaagttc tttttccacg tgcctatcac actgaactat caggccgcca attccccatc
     5641 taagttcaac cagagggtga atgcctacct gaaggagcac cccgagacac ctatcatcgg
     5701 catcgatcgg ggcgagagaa acctgatcta tatcacagtg atcgactcca ccggcaagat
     5761 cctggagcag cggagcctga acaccatcca gcagtttgat taccagaaga agctggacaa
     5821 cagggagaag gagagggtgg cagcaaggca ggcctggtct gtggtgggca caatcaagga
     5881 tctgaagcag ggctatctga gccaggtcat ccacgagatc gtggacctga tgatccacta
     5941 ccaggccgtg gtggtgctgg agaacctgaa tttcggcttt aagagcaaga ggaccggcat
     6001 cgccgagaag gccgtgtacc agcagttcga gaagatgctg atcgataagc tgaattgcct
     6061 ggtgctgaag gactatccag cagagaaagt gggaggcgtg ctgaacccat accagctgac
     6121 agaccagttc acctcctttg ccaagatggg cacccagtct ggcttcctgt tttacgtgcc
     6181 tgccccatat acatctaaga tcgatcccct gaccggcttc gtggacccct tcgtgtggaa
     6241 aaccatcaag aatcacgaga gccgcaagca cttcctggag ggcttcgact ttctgcacta
     6301 cgacgtgaaa accggcgact tcatcctgca ctttaagatg aacagaaatc tgtccttcca
     6361 gaggggcctg cccggcttta tgcctgcatg ggatatcgtg ttcgagaaga acgagacaca
     6421 gtttgacgcc aagggcaccc ctttcatcgc cggcaagaga atcgtgccag tgatcgagaa
     6481 tcacagattc accggcagat accgggacct gtatcctgcc aacgagctga tcgccctgct
     6541 ggaggagaag ggcatcgtgt tcagggatgg ctccaacatc ctgccaaagc tgctggagaa
     6601 tgacgattct cacgccatcg acaccatggt ggccctgatc cgcagcgtgc tgcagatgcg
     6661 gaactccaat gccgccacag gcgaggacta tatcaacagc cccgtgcgcg atctgaatgg
     6721 cgtgtgcttc gactcccggt ttcagaaccc agagtggccc atggacgccg atgccaatgg
     6781 cgcctaccac atcgccctga agggccagct gctgctgaat cacctgaagg agagcaagga
     6841 tctgaagctg cagaacggca tctccaatca ggactggctg gcctacatcc aggagctgcg
     6901 caacaaaagg ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa agggatccta
     6961 cccatacgat gttccagatt acgcttatcc ctacgacgtg cctgattatg catacccata
     7021 tgatgtcccc gactatgccg gatccggcgc aacaaacttc tctctgctga aacaagccgg
     7081 agatgtcgaa gagaatcctg gaccgaccga gtacaagccc acggtgcgcc tcgccacccg
     7141 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     7201 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     7261 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     7321 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     7381 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     7441 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggag tctcgcccga
     7501 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     7561 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     7621 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     7681 gacccgcaag cccggtgcct gaacgcgtta agtcgacaat caacctctgg attacaaaat
     7741 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     7801 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     7861 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     7921 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     7981 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     8041 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     8101 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     8161 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     8221 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     8281 gatctccctt tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct
     8341 gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa
     8401 cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc
     8461 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
     8521 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
     8581 cccttttagt cagtgtggaa aatctctagc agggcccgtt taaacccgct gatcagcctc
     8641 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     8701 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     8761 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     8821 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     8881 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     8941 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     9001 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     9061 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     9121 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     9181 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     9241 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     9301 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     9361 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     9421 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     9481 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     9541 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     9601 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     9661 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     9721 gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     9781 aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg
     9841 acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg
     9901 aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg
     9961 accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt
    10021 acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga
    10081 ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact
    10141 gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg
    10201 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
    10261 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
    10321 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
    10381 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
    10441 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
    10501 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
    10561 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
    10621 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
    10681 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
    10741 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
    10801 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
    10861 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
    10921 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
    10981 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
    11041 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
    11101 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
    11161 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
    11221 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
    11281 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
    11341 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
    11401 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
    11461 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
    11521 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
    11581 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
    11641 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
    11701 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
    11761 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
    11821 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
    11881 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
    11941 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
    12001 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
    12061 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
    12121 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
    12181 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
    12241 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
    12301 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
    12361 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
    12421 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
    12481 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
    12541 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
    12601 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
    12661 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
    12721 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
    12781 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
    12841 tgtacgggcc agatatacgc gtt
//