LOCUS       Exported               13363 bp ds-DNA     circular SYN 17-FEB-2022
DEFINITION  CasRx (NLS-RfxCas13d-NLS) with 2A-EGFP for RNA targeting.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13363)
  AUTHORS   Konermann S, Lotfy P, Brideau NJ, Oki J, Shokhirev MN, Hsu PD
  TITLE     Transcriptome Engineering with RNA-Targeting Type VI-D CRISPR 
            Effectors.
  JOURNAL   Cell. 2018 Mar 8. pii: S0092-8674(18)30207-1. doi: 
            10.1016/j.cell.2018.02.033.
  PUBMED    29551272
REFERENCE   2  (bases 1 to 13363)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jan 11, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..13363
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    61..178
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        303..1481
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          534..1472
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1429..1449
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             1500..1520
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1530..4408
                     /codon_start=1
                     /product="RNA-targeting Type VI CRISPR protein from 
                     Ruminococcus flavefaciens strain XPD3002 (Konermann et al.,
                     2018)"
                     /label=RfxCas13d
                     /translation="IEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDSI
                     RSVNEGEAFSAEMADKNAGYKIGNAKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLEK
                     RYFGESADGNDNICIQVIHNILDIEKILAEYITNAAYAVNNISGLDKDIIGFGKFSTVY
                     TYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFSKEGRNYIIN
                     YGNECYDILALLSGLRHWVVHNNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITNE
                     LTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRKD
                     MSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFVI
                     NLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPAG
                     RDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAFF
                     KDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDEN
                     GNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQK
                     KQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGREN
                     AEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLEE
                     KGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKKA
                     EEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFRNKAVHLEVARYVHA
                     YINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCVP
                     FGYCIPRFKNLSIEALFDRNEAAKFDKEK"
     CDS             4437..4457
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4482..4535
                     /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             4536..4931
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             4953..5651
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     misc_feature    5676..6264
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(6147..6158)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6267..6283)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6789..6969
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6995..7012)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7001..7225
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7271..7699
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7358..7377)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7568..7589
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7708..7728)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7713..8042
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7893..8028
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7955..7974
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8090..8137
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8156..8530
                     /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    8660..8781
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8697..8716)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8751..8770
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8830..8846)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8830..8846)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8843..8865)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8854..8870
                     /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(8878..8908)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8923..8944
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9061..9078)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9232..9820)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9312..9331)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9991..10851)
                     /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     10614..10633
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10852..10956)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(11031..11050)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        11222..11601
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11603..11801
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     11756..11776
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             11819..11999
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    12046..12171
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12664..12897
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             13082..13126
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
ORIGIN
        1 gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa atggcagtat tcatccacaa
       61 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat
      121 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg
      181 ggtttattac agggacagca gagatccagt ttggttaatt aatgcaaaga tggataaagt
      241 tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa
      301 ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
      361 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
      421 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
      481 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
      541 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
      601 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
      661 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
      721 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
      781 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
      841 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
      901 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
      961 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
     1021 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
     1081 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
     1141 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     1201 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     1261 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     1321 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     1381 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     1441 cagtggttca aagttttttt cttccatttc aggtgtcgtg acgtacggcc accatgagcc
     1501 ccaagaagaa gagaaaggtg gaggccagca tcgaaaaaaa aaagtccttc gccaagggca
     1561 tgggcgtgaa gtccacactc gtgtccggct ccaaagtgta catgacaacc ttcgccgaag
     1621 gcagcgacgc caggctggaa aagatcgtgg agggcgacag catcaggagc gtgaatgagg
     1681 gcgaggcctt cagcgctgaa atggccgata aaaacgccgg ctataagatc ggcaacgcca
     1741 aattcagcca tcctaagggc tacgccgtgg tggctaacaa ccctctgtat acaggacccg
     1801 tccagcagga tatgctcggc ctgaaggaaa ctctggaaaa gaggtacttc ggcgagagcg
     1861 ctgatggcaa tgacaatatt tgtatccagg tgatccataa catcctggac attgaaaaaa
     1921 tcctcgccga atacattacc aacgccgcct acgccgtcaa caatatctcc ggcctggata
     1981 aggacattat tggattcggc aagttctcca cagtgtatac ctacgacgaa ttcaaagacc
     2041 ccgagcacca tagggccgct ttcaacaata acgataagct catcaacgcc atcaaggccc
     2101 agtatgacga gttcgacaac ttcctcgata accccagact cggctatttc ggccaggcct
     2161 ttttcagcaa ggagggcaga aattacatca tcaattacgg caacgaatgc tatgacattc
     2221 tggccctcct gagcggactg aggcactggg tggtccataa caacgaagaa gagtccagga
     2281 tctccaggac ctggctctac aacctcgata agaacctcga caacgaatac atctccaccc
     2341 tcaactacct ctacgacagg atcaccaatg agctgaccaa ctccttctcc aagaactccg
     2401 ccgccaacgt gaactatatt gccgaaactc tgggaatcaa ccctgccgaa ttcgccgaac
     2461 aatatttcag attcagcatt atgaaagagc agaaaaacct cggattcaat atcaccaagc
     2521 tcagggaagt gatgctggac aggaaggata tgtccgagat caggaaaaat cataaggtgt
     2581 tcgactccat caggaccaag gtctacacca tgatggactt tgtgatttat aggtattaca
     2641 tcgaagagga tgccaaggtg gctgccgcca ataagtccct ccccgataat gagaagtccc
     2701 tgagcgagaa ggatatcttt gtgattaacc tgaggggctc cttcaacgac gaccagaagg
     2761 atgccctcta ctacgatgaa gctaatagaa tttggagaaa gctcgaaaat atcatgcaca
     2821 acatcaagga atttagggga aacaagacaa gagagtataa gaagaaggac gcccctagac
     2881 tgcccagaat cctgcccgct ggccgtgatg tttccgcctt cagcaaactc atgtatgccc
     2941 tgaccatgtt cctggatggc aaggagatca acgacctcct gaccaccctg attaataaat
     3001 tcgataacat ccagagcttc ctgaaggtga tgcctctcat cggagtcaac gctaagttcg
     3061 tggaggaata cgcctttttc aaagactccg ccaagatcgc cgatgagctg aggctgatca
     3121 agtccttcgc tagaatggga gaacctattg ccgatgccag gagggccatg tatatcgacg
     3181 ccatccgtat tttaggaacc aacctgtcct atgatgagct caaggccctc gccgacacct
     3241 tttccctgga cgagaacgga aacaagctca agaaaggcaa gcacggcatg agaaatttca
     3301 ttattaataa cgtgatcagc aataaaaggt tccactacct gatcagatac ggtgatcctg
     3361 cccacctcca tgagatcgcc aaaaacgagg ccgtggtgaa gttcgtgctc ggcaggatcg
     3421 ctgacatcca gaaaaaacag ggccagaacg gcaagaacca gatcgacagg tactacgaaa
     3481 cttgtatcgg aaaggataag ggcaagagcg tgagcgaaaa ggtggacgct ctcacaaaga
     3541 tcatcaccgg aatgaactac gaccaattcg acaagaaaag gagcgtcatt gaggacaccg
     3601 gcagggaaaa cgccgagagg gagaagttta aaaagatcat cagcctgtac ctcaccgtga
     3661 tctaccacat cctcaagaat attgtcaata tcaacgccag gtacgtcatc ggattccatt
     3721 gcgtcgagcg tgatgctcaa ctgtacaagg agaaaggcta cgacatcaat ctcaagaaac
     3781 tggaagagaa gggattcagc tccgtcacca agctctgcgc tggcattgat gaaactgccc
     3841 ccgataagag aaaggacgtg gaaaaggaga tggctgaaag agccaaggag agcattgaca
     3901 gcctcgagag cgccaacccc aagctgtatg ccaattacat caaatacagc gacgagaaga
     3961 aagccgagga gttcaccagg cagattaaca gggagaaggc caaaaccgcc ctgaacgcct
     4021 acctgaggaa caccaagtgg aatgtgatca tcagggagga cctcctgaga attgacaaca
     4081 agacatgtac cctgttcaga aacaaggccg tccacctgga agtggccagg tatgtccacg
     4141 cctatatcaa cgacattgcc gaggtcaatt cctacttcca actgtaccat tacatcatgc
     4201 agagaattat catgaatgag aggtacgaga aaagcagcgg aaaggtgtcc gagtacttcg
     4261 acgctgtgaa tgacgagaag aagtacaacg ataggctcct gaaactgctg tgtgtgcctt
     4321 tcggctactg tatccccagg tttaagaacc tgagcatcga ggccctgttc gataggaacg
     4381 aggccgccaa gttcgacaag gagaaaaaga aggtgtccgg caattccgga tccggaccta
     4441 agaaaaagag gaaggtggcg gccgctggat ccggcagtgg agagggcaga ggaagtctgc
     4501 taacatgcgg tgacgtcgag gagaatcctg gcccaatggc caagcctttg tctcaagaag
     4561 aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact
     4621 acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat
     4681 atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg
     4741 cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc
     4801 cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga
     4861 aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg
     4921 tgtgggaggg cgctagcggc agtggaagtg gcagcgagct gattaaggag aacatgcaca
     4981 tgaagctcta catggagggc accgtggaca accatcactt caagtgcaca tccgagggcg
     5041 aaggcaagcc ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc
     5101 tccccttcgc cttcgacatc ctggctacta gcttcctcta cggcagcaag accttcatca
     5161 accacaccca gggcatcccc gacttcttca agcagtcctt ccctgagggc ttcacatggg
     5221 agagagtcac cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc
     5281 aggacggctg cctcatctac aacgtcaaga tcagaggggt gaacttcaca tccaacggcc
     5341 ctgtgatgca gaagaaaaca ctcggctggg aggccttcac cgaaacgctg taccccgctg
     5401 acggcggcct ggaaggcaga aacgacatgg ccctgaagct cgtgggcggg agccatctga
     5461 tcgcaaacat caagaccaca tatagatcca agaaacccgc taagaacctc aagatgcctg
     5521 gcgtctacta tgtggactac agactggaaa gaatcaagga ggccaacaac gagacctacg
     5581 tcgagcagca cgaggtggca gtggccagat actgcgacct ccctagcaaa ctggggcaca
     5641 agcttaatta agaattcgat atcaagctta tcggtaatca acctctggat tacaaaattt
     5701 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     5761 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     5821 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     5881 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     5941 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     6001 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     6061 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     6121 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     6181 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     6241 tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgagaccta gaaaaacatg
     6301 gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac
     6361 aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt
     6421 acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa
     6481 ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact
     6541 tccctgattg gcagaactac acaccagggc cagggatcag atatccactg acctttggat
     6601 ggtgctacaa gctagtacca gttgagcaag agaaggtaga agaagccaat gaaggagaga
     6661 acacccgctt gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtat
     6721 tagagtggag gtttgacagc cgcctagcat ttcatcacat ggcccgagag ctgcatccgg
     6781 actgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag
     6841 ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc
     6901 gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa
     6961 tctctagcag ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     7021 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     7081 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     7141 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     7201 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     7261 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     7321 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     7381 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     7441 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     7501 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     7561 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     7621 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     7681 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     7741 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     7801 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     7861 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     7921 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     7981 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     8041 aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta
     8101 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc
     8161 caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt
     8221 ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg ccggtgtggt
     8281 ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc cggacaacac
     8341 cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt cggaggtcgt
     8401 gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg agcagccgtg
     8461 ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg tggccgagga
     8521 gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     8581 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     8641 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     8701 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     8761 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     8821 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     8881 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     8941 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     9001 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     9061 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     9121 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     9181 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     9241 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     9301 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     9361 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     9421 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     9481 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     9541 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     9601 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     9661 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     9721 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     9781 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     9841 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     9901 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     9961 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
    10021 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
    10081 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
    10141 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
    10201 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
    10261 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
    10321 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
    10381 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
    10441 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
    10501 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
    10561 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
    10621 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
    10681 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
    10741 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
    10801 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
    10861 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
    10921 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
    10981 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
    11041 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
    11101 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
    11161 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
    11221 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
    11281 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
    11341 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
    11401 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
    11461 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
    11521 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
    11581 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
    11641 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
    11701 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
    11761 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg
    11821 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
    11881 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
    11941 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
    12001 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
    12061 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
    12121 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
    12181 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
    12241 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
    12301 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
    12361 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
    12421 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
    12481 aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga
    12541 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
    12601 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
    12661 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
    12721 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
    12781 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
    12841 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
    12901 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
    12961 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
    13021 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
    13081 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
    13141 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
    13201 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
    13261 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
    13321 agaagaaggt ggagagagag acagagacag atccattcga tta
//
