LOCUS       sequence-205481-       12990 bp    DNA     circular SYN 15-DEC-2025
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 12990)
  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 12990)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2018 
            Mar 8. pii: S0092-8674(18)30207-1. doi: 10.1016/j.cell.2018.02.033."
COMMENT     Sequence Label: pXR001: EF1a-CasRx-2A-EGFP
FEATURES             Location/Qualifiers
     source          1..12990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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"
     misc_feature    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2682..3859
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2913..3850
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3807..3827
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3878..3898
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3908..6805
                     /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
                     FGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGNS"
     CDS             6815..6835
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6845..6871
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6887..6940
                     /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             join(6941..6943,6944..7657)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(6941..6943,6944..7654)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6983..7004)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7244..7263)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7591..7612
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    7682..8270
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7735..7755)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8153..8164)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8273..8289)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8274..8290)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8795..8975
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9001..9018)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9007..9231
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9277..9705
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9364..9383)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9574..9595
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9714..9734)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9719..10048
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9899..10034
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9961..9980
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10096..10143
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10162..10536
                     /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    10666..10787
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10703..10722)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10757..10776
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10836..10852)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10836..10852)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10849..10871)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10860..10876
                     /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(join(10884..10890,10891..10908,10909..10914))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10929..10950
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11067..11084)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11238..11826)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11318..11337)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11997..12788,12789..12857))
                     /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     12620..12639
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12858..12962)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta atgcaaagat ggataaagtt ttaaacagag aggaatcttt
     2641 gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt
     2701 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     2761 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     2821 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
     2881 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
     2941 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccact ggctgcagta
     3001 cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc
     3061 ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg
     3121 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat
     3181 ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc
     3241 gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg
     3301 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg
     3361 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat
     3421 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg
     3481 gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg
     3541 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg
     3601 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag
     3661 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg
     3721 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct
     3781 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct
     3841 tccatttcag gtgtcgtgac gtacggccac catgagcccc aagaagaaga gaaaggtgga
     3901 ggccagcatc gaaaaaaaaa agtccttcgc caagggcatg ggcgtgaagt ccacactcgt
     3961 gtccggctcc aaagtgtaca tgacaacctt cgccgaaggc agcgacgcca ggctggaaaa
     4021 gatcgtggag ggcgacagca tcaggagcgt gaatgagggc gaggccttca gcgctgaaat
     4081 ggccgataaa aacgccggct ataagatcgg caacgccaaa ttcagccatc ctaagggcta
     4141 cgccgtggtg gctaacaacc ctctgtatac aggacccgtc cagcaggata tgctcggcct
     4201 gaaggaaact ctggaaaaga ggtacttcgg cgagagcgct gatggcaatg acaatatttg
     4261 tatccaggtg atccataaca tcctggacat tgaaaaaatc ctcgccgaat acattaccaa
     4321 cgccgcctac gccgtcaaca atatctccgg cctggataag gacattattg gattcggcaa
     4381 gttctccaca gtgtatacct acgacgaatt caaagacccc gagcaccata gggccgcttt
     4441 caacaataac gataagctca tcaacgccat caaggcccag tatgacgagt tcgacaactt
     4501 cctcgataac cccagactcg gctatttcgg ccaggccttt ttcagcaagg agggcagaaa
     4561 ttacatcatc aattacggca acgaatgcta tgacattctg gccctcctga gcggactgag
     4621 gcactgggtg gtccataaca acgaagaaga gtccaggatc tccaggacct ggctctacaa
     4681 cctcgataag aacctcgaca acgaatacat ctccaccctc aactacctct acgacaggat
     4741 caccaatgag ctgaccaact ccttctccaa gaactccgcc gccaacgtga actatattgc
     4801 cgaaactctg ggaatcaacc ctgccgaatt cgccgaacaa tatttcagat tcagcattat
     4861 gaaagagcag aaaaacctcg gattcaatat caccaagctc agggaagtga tgctggacag
     4921 gaaggatatg tccgagatca ggaaaaatca taaggtgttc gactccatca ggaccaaggt
     4981 ctacaccatg atggactttg tgatttatag gtattacatc gaagaggatg ccaaggtggc
     5041 tgccgccaat aagtccctcc ccgataatga gaagtccctg agcgagaagg atatctttgt
     5101 gattaacctg aggggctcct tcaacgacga ccagaaggat gccctctact acgatgaagc
     5161 taatagaatt tggagaaagc tcgaaaatat catgcacaac atcaaggaat ttaggggaaa
     5221 caagacaaga gagtataaga agaaggacgc ccctagactg cccagaatcc tgcccgctgg
     5281 ccgtgatgtt tccgccttca gcaaactcat gtatgccctg accatgttcc tggatggcaa
     5341 ggagatcaac gacctcctga ccaccctgat taataaattc gataacatcc agagcttcct
     5401 gaaggtgatg cctctcatcg gagtcaacgc taagttcgtg gaggaatacg cctttttcaa
     5461 agactccgcc aagatcgccg atgagctgag gctgatcaag tccttcgcta gaatgggaga
     5521 acctattgcc gatgccagga gggccatgta tatcgacgcc atccgtattt taggaaccaa
     5581 cctgtcctat gatgagctca aggccctcgc cgacaccttt tccctggacg agaacggaaa
     5641 caagctcaag aaaggcaagc acggcatgag aaatttcatt attaataacg tgatcagcaa
     5701 taaaaggttc cactacctga tcagatacgg tgatcctgcc cacctccatg agatcgccaa
     5761 aaacgaggcc gtggtgaagt tcgtgctcgg caggatcgct gacatccaga aaaaacaggg
     5821 ccagaacggc aagaaccaga tcgacaggta ctacgaaact tgtatcggaa aggataaggg
     5881 caagagcgtg agcgaaaagg tggacgctct cacaaagatc atcaccggaa tgaactacga
     5941 ccaattcgac aagaaaagga gcgtcattga ggacaccggc agggaaaacg ccgagaggga
     6001 gaagtttaaa aagatcatca gcctgtacct caccgtgatc taccacatcc tcaagaatat
     6061 tgtcaatatc aacgccaggt acgtcatcgg attccattgc gtcgagcgtg atgctcaact
     6121 gtacaaggag aaaggctacg acatcaatct caagaaactg gaagagaagg gattcagctc
     6181 cgtcaccaag ctctgcgctg gcattgatga aactgccccc gataagagaa aggacgtgga
     6241 aaaggagatg gctgaaagag ccaaggagag cattgacagc ctcgagagcg ccaaccccaa
     6301 gctgtatgcc aattacatca aatacagcga cgagaagaaa gccgaggagt tcaccaggca
     6361 gattaacagg gagaaggcca aaaccgccct gaacgcctac ctgaggaaca ccaagtggaa
     6421 tgtgatcatc agggaggacc tcctgagaat tgacaacaag acatgtaccc tgttcagaaa
     6481 caaggccgtc cacctggaag tggccaggta tgtccacgcc tatatcaacg acattgccga
     6541 ggtcaattcc tacttccaac tgtaccatta catcatgcag agaattatca tgaatgagag
     6601 gtacgagaaa agcagcggaa aggtgtccga gtacttcgac gctgtgaatg acgagaagaa
     6661 gtacaacgat aggctcctga aactgctgtg tgtgcctttc ggctactgta tccccaggtt
     6721 taagaacctg agcatcgagg ccctgttcga taggaacgag gccgccaagt tcgacaagga
     6781 gaaaaagaag gtgtccggca attccggatc cggacctaag aaaaagagga aggtggcggc
     6841 cgcttaccca tacgatgttc cagattacgc tgctagcggc agtggagagg gcagaggaag
     6901 tctgctaaca tgcggtgacg tcgaggagaa tcctggccca gtgagcaagg gcgaggagct
     6961 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     7021 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     7081 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     7141 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     7201 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     7261 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     7321 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     7381 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     7441 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     7501 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     7561 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     7621 cgggatcact ctcggcatgg acgagctgta caagtaagaa ttcgatatca agcttatcga
     7681 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     7741 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     7801 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     7861 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     7921 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     7981 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     8041 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     8101 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     8161 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     8221 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     8281 tcgacctcga gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa
     8341 tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc
     8401 tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     8461 agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct
     8521 gtggatctac cacacacaag gctacttccc tgattggcag aactacacac cagggccagg
     8581 gatcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg agcaagagaa
     8641 ggtagaagaa gccaatgaag gagagaacac ccgcttgtta caccctgtga gcctgcatgg
     8701 gatggatgac ccggagagag aagtattaga gtggaggttt gacagccgcc tagcatttca
     8761 tcacatggcc cgagagctgc atccggactg tactgggtct ctctggttag accagatctg
     8821 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     8881 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     8941 cagacccttt tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag
     9001 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     9061 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     9121 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     9181 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     9241 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     9301 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     9361 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     9421 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     9481 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     9541 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     9601 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     9661 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     9721 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     9781 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     9841 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     9901 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     9961 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
    10021 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
    10081 atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac
    10141 gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg
    10201 cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc
    10261 gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc
    10321 caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag
    10381 ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc
    10441 atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc
    10501 aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc
    10561 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
    10621 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
    10681 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
    10741 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
    10801 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
    10861 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
    10921 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
    10981 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
    11041 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
    11101 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
    11161 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
    11221 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
    11281 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
    11341 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
    11401 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
    11461 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
    11521 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
    11581 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
    11641 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
    11701 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
    11761 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
    11821 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
    11881 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
    11941 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
    12001 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
    12061 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
    12121 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
    12181 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
    12241 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
    12301 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
    12361 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
    12421 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
    12481 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
    12541 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
    12601 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
    12661 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
    12721 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
    12781 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
    12841 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
    12901 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
    12961 cgcacatttc cccgaaaagt gccacctgac
//
