pY094

LOCUS       pY094               10463 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  Expresses huAsCpf1-T2A-GFP and crRNA guide.
ACCESSION   .
VERSION     .
KEYWORDS    pY094
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10463)
  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 10463)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10463
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        170..410
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     170..190
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     341..360
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        538..917
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        918..1121
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1072..1092
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1166..1185
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1166..1184
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1232..5152
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND
                     HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN
                     AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD
                     KFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF
                     ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL
                     NLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL
                     RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG
                     KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL
                     KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA
                     RNYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYK
                     ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP
                     LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID
                     LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKG
                     HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK
                     DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFF
                     HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQR
                     SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV
                     VVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ
                     FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD
                     VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN
                     HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM
                     RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES
                     KDLKLQNGISNQDWLAYIQELRN"
     regulatory      4872..4881
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             5153..5200
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5216..5269
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             5270..5986
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5312..5333)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5573..5592)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5920..5941
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6057..6074)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6063..6287
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6333..6761
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6420..6439)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6630..6651
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6770..6790)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6775..7104
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6955..7090
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7017..7036
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             7171..7965
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7225..7244)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7835..7854
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8139..8260
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8176..8195)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8230..8249
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8309..8325)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8309..8325)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8322..8344)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8333..8349
                     /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(8357..8387)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8402..8423
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8540..8557)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8711..9296)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8791..8810)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9467..10327)
                     /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     10090..10109
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10328..10432)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga cttaattaag agggcctatt
      181 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat
      241 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt
      301 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt
      361 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgt
      421 aatttctact cttgtagatg gagacgatat atcgtctctt tttcaattgc atgaagaatc
      481 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      541 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      601 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      661 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      721 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      781 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      841 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      901 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      961 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     1021 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     1081 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac
     1141 ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct
     1201 agcgtttaaa cttaagcttg gtaccgccac catgacacag ttcgagggct ttaccaacct
     1261 gtatcaggtg agcaagacac tgcggtttga gctgatccca cagggcaaga ccctgaagca
     1321 catccaggag cagggcttca tcgaggagga caaggcccgc aatgatcact acaaggagct
     1381 gaagcccatc atcgatcgga tctacaagac ctatgccgac cagtgcctgc agctggtgca
     1441 gctggattgg gagaacctga gcgccgccat cgactcctat agaaaggaga aaaccgagga
     1501 gacaaggaac gccctgatcg aggagcaggc cacatatcgc aatgccatcc acgactactt
     1561 catcggccgg acagacaacc tgaccgatgc catcaataag agacacgccg agatctacaa
     1621 gggcctgttc aaggccgagc tgtttaatgg caaggtgctg aagcagctgg gcaccgtgac
     1681 cacaaccgag cacgagaacg ccctgctgcg gagcttcgac aagtttacaa cctacttctc
     1741 cggcttttat gagaacagga agaacgtgtt cagcgccgag gatatcagca cagccatccc
     1801 acaccgcatc gtgcaggaca acttccccaa gtttaaggag aattgtcaca tcttcacacg
     1861 cctgatcacc gccgtgccca gcctgcggga gcactttgag aacgtgaaga aggccatcgg
     1921 catcttcgtg agcacctcca tcgaggaggt gttttccttc cctttttata accagctgct
     1981 gacacagacc cagatcgacc tgtataacca gctgctggga ggaatctctc gggaggcagg
     2041 caccgagaag atcaagggcc tgaacgaggt gctgaatctg gccatccaga agaatgatga
     2101 gacagcccac atcatcgcct ccctgccaca cagattcatc cccctgttta agcagatcct
     2161 gtccgatagg aacaccctgt ctttcatcct ggaggagttt aagagcgacg aggaagtgat
     2221 ccagtccttc tgcaagtaca agacactgct gagaaacgag aacgtgctgg agacagccga
     2281 ggccctgttt aacgagctga acagcatcga cctgacacac atcttcatca gccacaagaa
     2341 gctggagaca atcagcagcg ccctgtgcga ccactgggat acactgagga atgccctgta
     2401 tgagcggaga atctccgagc tgacaggcaa gatcaccaag tctgccaagg agaaggtgca
     2461 gcgcagcctg aagcacgagg atatcaacct gcaggagatc atctctgccg caggcaagga
     2521 gctgagcgag gccttcaagc agaaaaccag cgagatcctg tcccacgcac acgccgccct
     2581 ggatcagcca ctgcctacaa ccctgaagaa gcaggaggag aaggagatcc tgaagtctca
     2641 gctggacagc ctgctgggcc tgtaccacct gctggactgg tttgccgtgg atgagtccaa
     2701 cgaggtggac cccgagttct ctgcccggct gaccggcatc aagctggaga tggagccttc
     2761 tctgagcttc tacaacaagg ccagaaatta tgccaccaag aagccctact ccgtggagaa
     2821 gttcaagctg aactttcaga tgcctacact ggcctctggc tgggacgtga ataaggagaa
     2881 gaacaatggc gccatcctgt ttgtgaagaa cggcctgtac tatctgggca tcatgccaaa
     2941 gcagaagggc aggtataagg ccctgagctt cgagcccaca gagaaaacca gcgagggctt
     3001 tgataagatg tactatgact acttccctga tgccgccaag atgatcccaa agtgcagcac
     3061 ccagctgaag gccgtgacag cccactttca gacccacaca acccccatcc tgctgtccaa
     3121 caatttcatc gagcctctgg agatcacaaa ggagatctac gacctgaaca atcctgagaa
     3181 ggagccaaag aagtttcaga cagcctacgc caagaaaacc ggcgaccaga agggctacag
     3241 agaggccctg tgcaagtgga tcgacttcac aagggatttt ctgtccaagt ataccaagac
     3301 aacctctatc gatctgtcta gcctgcggcc atcctctcag tataaggacc tgggcgagta
     3361 ctatgccgag ctgaatcccc tgctgtacca catcagcttc cagagaatcg ccgagaagga
     3421 gatcatggat gccgtggaga caggcaagct gtacctgttc cagatctata acaaggactt
     3481 tgccaagggc caccacggca agcctaatct gcacacactg tattggaccg gcctgttttc
     3541 tccagagaac ctggccaaga caagcatcaa gctgaatggc caggccgagc tgttctaccg
     3601 ccctaagtcc aggatgaaga ggatggcaca ccggctggga gagaagatgc tgaacaagaa
     3661 gctgaaggat cagaaaaccc caatccccga caccctgtac caggagctgt acgactatgt
     3721 gaatcacaga ctgtcccacg acctgtctga tgaggccagg gccctgctgc ccaacgtgat
     3781 caccaaggag gtgtctcacg agatcatcaa ggataggcgc tttaccagcg acaagttctt
     3841 tttccacgtg cctatcacac tgaactatca ggccgccaat tccccatcta agttcaacca
     3901 gagggtgaat gcctacctga aggagcaccc cgagacacct atcatcggca tcgatcgggg
     3961 cgagagaaac ctgatctata tcacagtgat cgactccacc ggcaagatcc tggagcagcg
     4021 gagcctgaac accatccagc agtttgatta ccagaagaag ctggacaaca gggagaagga
     4081 gagggtggca gcaaggcagg cctggtctgt ggtgggcaca atcaaggatc tgaagcaggg
     4141 ctatctgagc caggtcatcc acgagatcgt ggacctgatg atccactacc aggccgtggt
     4201 ggtgctggag aacctgaatt tcggctttaa gagcaagagg accggcatcg ccgagaaggc
     4261 cgtgtaccag cagttcgaga agatgctgat cgataagctg aattgcctgg tgctgaagga
     4321 ctatccagca gagaaagtgg gaggcgtgct gaacccatac cagctgacag accagttcac
     4381 ctcctttgcc aagatgggca cccagtctgg cttcctgttt tacgtgcctg ccccatatac
     4441 atctaagatc gatcccctga ccggcttcgt ggaccccttc gtgtggaaaa ccatcaagaa
     4501 tcacgagagc cgcaagcact tcctggaggg cttcgacttt ctgcactacg acgtgaaaac
     4561 cggcgacttc atcctgcact ttaagatgaa cagaaatctg tccttccaga ggggcctgcc
     4621 cggctttatg cctgcatggg atatcgtgtt cgagaagaac gagacacagt ttgacgccaa
     4681 gggcacccct ttcatcgccg gcaagagaat cgtgccagtg atcgagaatc acagattcac
     4741 cggcagatac cgggacctgt atcctgccaa cgagctgatc gccctgctgg aggagaaggg
     4801 catcgtgttc agggatggct ccaacatcct gccaaagctg ctggagaatg acgattctca
     4861 cgccatcgac accatggtgg ccctgatccg cagcgtgctg cagatgcgga actccaatgc
     4921 cgccacaggc gaggactata tcaacagccc cgtgcgcgat ctgaatggcg tgtgcttcga
     4981 ctcccggttt cagaacccag agtggcccat ggacgccgat gccaatggcg cctaccacat
     5041 cgccctgaag ggccagctgc tgctgaatca cctgaaggag agcaaggatc tgaagctgca
     5101 gaacggcatc tccaatcagg actggctggc ctacatccag gagctgcgca acaaaaggcc
     5161 ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag ggatccggca gtggagaggg
     5221 cagaggaagt ctgctaacat gcggtgacgt cgaggagaat cctggcccag tgagcaaggg
     5281 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     5341 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     5401 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     5461 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     5521 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     5581 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     5641 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     5701 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     5761 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     5821 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     5881 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     5941 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagaat tctgcagata
     6001 tccagcacag tggcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc
     6061 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     6121 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     6181 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     6241 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     6301 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     6361 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     6421 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     6481 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     6541 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     6601 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     6661 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     6721 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     6781 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     6841 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     6901 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     6961 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     7021 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     7081 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     7141 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     7201 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     7261 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     7321 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     7381 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     7441 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     7501 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     7561 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     7621 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     7681 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     7741 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     7801 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     7861 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     7921 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     7981 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     8041 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     8101 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     8161 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     8221 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     8281 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     8341 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     8401 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     8461 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     8521 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     8581 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     8641 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     8701 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     8761 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     8821 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     8881 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     8941 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     9001 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     9061 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     9121 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     9181 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     9241 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     9301 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     9361 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     9421 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     9481 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     9541 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     9601 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     9661 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     9721 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     9781 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     9841 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     9901 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     9961 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
    10021 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
    10081 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
    10141 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
    10201 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
    10261 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
    10321 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
    10381 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
    10441 cccgaaaagt gccacctgac gtc
//