LOCUS       sequence-318593-       11728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses RfxCas13d in mammalian cells, localized in the cytoplasm. 
            For cloning of guide RNAs compatible with RfxCas13d. Contains a 5' 
            direct repeat. Clone using BsmBI. F overhang aaac. R overhang aaaa.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11728)
  AUTHORS   Wessels HH, Mendez-Mancilla A, Guo X, Legut M, Daniloski Z, Sanjana 
            NE
  TITLE     Massively parallel Cas13 screens reveal principles for guide RNA 
            design.
  JOURNAL   Nat Biotechnol. 2020 Jun;38(6):722-727. doi: 
            10.1038/s41587-020-0456-9. Epub 2020 Mar 16.
  PUBMED    32518401
REFERENCE   2  (bases 1 to 11728)
  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"; doi: 
            "10.1038/s41587-020-0456-9"; journalName: "Nat Biotechnol"; date: 
            "2020-06"; volume: "38"; issue: "6"; pages: "722-727"
COMMENT     Sequence Label: pLentiRNACRISPR_006 -
            hU6-DR_BsmBI-EFS-RfxCas1...
FEATURES             Location/Qualifiers
     source          1..11728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(58..77)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        249..628
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        630..828
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     783..803
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             846..1026
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1073..1198
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1691..1924
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2109..2153
                     /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    2451..2568
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2619..2867
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2619..2639
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2790..2809
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2869..2898
                     /label=RfxCas13d DR30
                     /note="5' processed 30-base direct repeat for an RfxCas13d 
                     gRNA (Konermann et al., 2018)"
     promoter        2977..3188
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             3213..6110
                     /codon_start=1
                     /product="RNA-targeting Type VI CRISPR protein from 
                     Ruminococcus flavefaciens strain XPD3002 (Konermann et al.,
                     2018)"
                     /label=RfxCas13d
                     /translation="MIEKKKSFAKGMGVKSTLVSGSKVYMTTFAEGSDARLEKIVEGDS
                     IRSVNEGEAFSAEMADKNAGYKIGNAKFSHPKGYAVVANNPLYTGPVQQDMLGLKETLE
                     KRYFGESADGNDNICIQVIHNILDIEKILAEYITNAAYAVNNISGLDKDIIGFGKFSTV
                     YTYDEFKDPEHHRAAFNNNDKLINAIKAQYDEFDNFLDNPRLGYFGQAFFSKEGRNYII
                     NYGNECYDILALLSGLRHWVVHNNEEESRISRTWLYNLDKNLDNEYISTLNYLYDRITN
                     ELTNSFSKNSAANVNYIAETLGINPAEFAEQYFRFSIMKEQKNLGFNITKLREVMLDRK
                     DMSEIRKNHKVFDSIRTKVYTMMDFVIYRYYIEEDAKVAAANKSLPDNEKSLSEKDIFV
                     INLRGSFNDDQKDALYYDEANRIWRKLENIMHNIKEFRGNKTREYKKKDAPRLPRILPA
                     GRDVSAFSKLMYALTMFLDGKEINDLLTTLINKFDNIQSFLKVMPLIGVNAKFVEEYAF
                     FKDSAKIADELRLIKSFARMGEPIADARRAMYIDAIRILGTNLSYDELKALADTFSLDE
                     NGNKLKKGKHGMRNFIINNVISNKRFHYLIRYGDPAHLHEIAKNEAVVKFVLGRIADIQ
                     KKQGQNGKNQIDRYYETCIGKDKGKSVSEKVDALTKIITGMNYDQFDKKRSVIEDTGRE
                     NAEREKFKKIISLYLTVIYHILKNIVNINARYVIGFHCVERDAQLYKEKGYDINLKKLE
                     EKGFSSVTKLCAGIDETAPDKRKDVEKEMAERAKESIDSLESANPKLYANYIKYSDEKK
                     AEEFTRQINREKAKTALNAYLRNTKWNVIIREDLLRIDNKTCTLFRNKAVHLEVARYVH
                     AYINDIAEVNSYFQLYHYIMQRIIMNERYEKSSGKVSEYFDAVNDEKKYNDRLLKLLCV
                     PFGYCIPRFKNLSIEALFDRNEAAKFDKEKKKVSGN"
     CDS             6117..6143
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             6162..6218
                     /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             6219..6815
                     /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     6712..6732
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6831..7419
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6884..6904)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7302..7313)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7491..7724
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7750..7767)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7756..7980
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8026..8454
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8113..8132)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8323..8344
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8463..8483)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        8468..8797
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8648..8783
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8710..8729
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8845..8892
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8911..9285
                     /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    9415..9536
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9452..9471)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9506..9525
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(9585..9601)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9585..9601)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9598..9620)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9609..9625
                     /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(9633..9639,9640..9657,9658..9663))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9678..9699
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9816..9833)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9987..10575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10067..10086)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10746..11537,11538..11606))
                     /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     11369..11388
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11607..11711)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
       61 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
      121 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
      181 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
      241 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
      301 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      361 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      421 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      481 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      541 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      601 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      661 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      721 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      781 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagcg cgttttgcct
      841 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
      901 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
      961 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     1021 ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg agctctctcg
     1081 acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc gactggtgag
     1141 tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg agagcgtcag
     1201 tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc cagggggaaa
     1261 gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac gattcgcagt
     1321 taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac agctacaacc
     1381 atcccttcag acaggatcag aagaacttag atcattatat aatacagtag caaccctcta
     1441 ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca agatagagga
     1501 agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc agacctggag
     1561 gaggagatat gagggacaat tggagaagtg aattatataa atataaagta gtaaaaattg
     1621 aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga gaaaaaagag
     1681 cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc actatgggcg
     1741 cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata gtgcagcagc
     1801 agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc acagtctggg
     1861 gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag gatcaacagc
     1921 tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg ccttggaatg
     1981 ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg atggagtggg
     2041 acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa tcgcaaaacc
     2101 agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt ttgtggaatt
     2161 ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata gtaggaggct
     2221 tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt aggcagggat
     2281 attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac aggcccgaag
     2341 gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta gtgaacggat
     2401 cggcactgcg tgcgccaatt ctgcagacaa atggcagtat tcatccacaa ttttaaaaga
     2461 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     2521 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac
     2581 agggacagca gagatccagt ttggttaatt aaggtaccga gggcctattt cccatgattc
     2641 cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg
     2701 taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt
     2761 ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt
     2821 atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccgaa cccctaccaa
     2881 ctggtcgggg tttgaaacgg agacggacgt ctcttttttt tgaattcgct agctaggtct
     2941 tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
     3001 tccccgagaa gttgggggga ggggtcggca attgatccgg tgcctagaga aggtggcgcg
     3061 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
     3121 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
     3181 gaacacagga ccggttctag agcgctgcca ccatgatcga aaaaaaaaag tccttcgcca
     3241 agggcatggg cgtgaagtcc acactcgtgt ccggctccaa agtgtacatg acaaccttcg
     3301 ccgaaggcag cgacgccagg ctggaaaaga tcgtggaggg cgacagcatc aggagcgtga
     3361 atgagggcga ggccttcagc gctgaaatgg ccgataaaaa cgccggctat aagatcggca
     3421 acgccaaatt cagccatcct aagggctacg ccgtggtggc taacaaccct ctgtatacag
     3481 gacccgtcca gcaggatatg ctcggcctga aggaaactct ggaaaagagg tacttcggcg
     3541 agagcgctga tggcaatgac aatatttgta tccaggtgat ccataacatc ctggacattg
     3601 aaaaaatcct cgccgaatac attaccaacg ccgcctacgc cgtcaacaat atctccggcc
     3661 tggataagga cattattgga ttcggcaagt tctccacagt gtatacctac gacgaattca
     3721 aagaccccga gcaccatagg gccgctttca acaataacga taagctcatc aacgccatca
     3781 aggcccagta tgacgagttc gacaacttcc tcgataaccc cagactcggc tatttcggcc
     3841 aggccttttt cagcaaggag ggcagaaatt acatcatcaa ttacggcaac gaatgctatg
     3901 acattctggc cctcctgagc ggactgaggc actgggtggt ccataacaac gaagaagagt
     3961 ccaggatctc caggacctgg ctctacaacc tcgataagaa cctcgacaac gaatacatct
     4021 ccaccctcaa ctacctctac gacaggatca ccaatgagct gaccaactcc ttctccaaga
     4081 actccgccgc caacgtgaac tatattgccg aaactctggg aatcaaccct gccgaattcg
     4141 ccgaacaata tttcagattc agcattatga aagagcagaa aaacctcgga ttcaatatca
     4201 ccaagctcag ggaagtgatg ctggacagga aggatatgtc cgagatcagg aaaaatcata
     4261 aggtgttcga ctccatcagg accaaggtct acaccatgat ggactttgtg atttataggt
     4321 attacatcga agaggatgcc aaggtggctg ccgccaataa gtccctcccc gataatgaga
     4381 agtccctgag cgagaaggat atctttgtga ttaacctgag gggctccttc aacgacgacc
     4441 agaaggatgc cctctactac gatgaagcta atagaatttg gagaaagctc gaaaatatca
     4501 tgcacaacat caaggaattt aggggaaaca agacaagaga gtataagaag aaggacgccc
     4561 ctagactgcc cagaatcctg cccgctggcc gtgatgtttc cgccttcagc aaactcatgt
     4621 atgccctgac catgttcctg gatggcaagg agatcaacga cctcctgacc accctgatta
     4681 ataaattcga taacatccag agcttcctga aggtgatgcc tctcatcgga gtcaacgcta
     4741 agttcgtgga ggaatacgcc tttttcaaag actccgccaa gatcgccgat gagctgaggc
     4801 tgatcaagtc cttcgctaga atgggagaac ctattgccga tgccaggagg gccatgtata
     4861 tcgacgccat ccgtatttta ggaaccaacc tgtcctatga tgagctcaag gccctcgccg
     4921 acaccttttc cctggacgag aacggaaaca agctcaagaa aggcaagcac ggcatgagaa
     4981 atttcattat taataacgtg atcagcaata aaaggttcca ctacctgatc agatacggtg
     5041 atcctgccca cctccatgag atcgccaaaa acgaggccgt ggtgaagttc gtgctcggca
     5101 ggatcgctga catccagaaa aaacagggcc agaacggcaa gaaccagatc gacaggtact
     5161 acgaaacttg tatcggaaag gataagggca agagcgtgag cgaaaaggtg gacgctctca
     5221 caaagatcat caccggaatg aactacgacc aattcgacaa gaaaaggagc gtcattgagg
     5281 acaccggcag ggaaaacgcc gagagggaga agtttaaaaa gatcatcagc ctgtacctca
     5341 ccgtgatcta ccacatcctc aagaatattg tcaatatcaa cgccaggtac gtcatcggat
     5401 tccattgcgt cgagcgtgat gctcaactgt acaaggagaa aggctacgac atcaatctca
     5461 agaaactgga agagaaggga ttcagctccg tcaccaagct ctgcgctggc attgatgaaa
     5521 ctgcccccga taagagaaag gacgtggaaa aggagatggc tgaaagagcc aaggagagca
     5581 ttgacagcct cgagagcgcc aaccccaagc tgtatgccaa ttacatcaaa tacagcgacg
     5641 agaagaaagc cgaggagttc accaggcaga ttaacaggga gaaggccaaa accgccctga
     5701 acgcctacct gaggaacacc aagtggaatg tgatcatcag ggaggacctc ctgagaattg
     5761 acaacaagac atgtaccctg ttcagaaaca aggccgtcca cctggaagtg gccaggtatg
     5821 tccacgccta tatcaacgac attgccgagg tcaattccta cttccaactg taccattaca
     5881 tcatgcagag aattatcatg aatgagaggt acgagaaaag cagcggaaag gtgtccgagt
     5941 acttcgacgc tgtgaatgac gagaagaagt acaacgatag gctcctgaaa ctgctgtgtg
     6001 tgcctttcgg ctactgtatc cccaggttta agaacctgag catcgaggcc ctgttcgata
     6061 ggaacgaggc cgccaagttc gacaaggaga aaaagaaggt gtccggcaat cttcaactgc
     6121 ctccacttga aagactgaca ctgggtagtt ccggatccgg cgcaacaaac ttctctctgc
     6181 tgaaacaagc cggagatgtc gaagagaatc ctggaccgac cgagtacaag cccacggtgc
     6241 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     6301 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     6361 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     6421 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     6481 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     6541 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     6601 gagtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     6661 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     6721 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     6781 gcacctggtg catgacccgc aagcccggtg cctgaacgcg ttaagtcgac aatcaacctc
     6841 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     6901 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     6961 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     7021 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     7081 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     7141 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     7201 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     7261 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     7321 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     7381 ctcagacgag tcggatctcc ctttgggccg cctccccgcg tcgactttaa gaccaatgac
     7441 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     7501 aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga
     7561 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     7621 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     7681 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagggccc gtttaaaccc
     7741 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     7801 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     7861 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     7921 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     7981 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     8041 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     8101 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     8161 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     8221 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     8281 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     8341 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     8401 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     8461 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     8521 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     8581 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     8641 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     8701 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     8761 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     8821 cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta tatcggcata
     8881 gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc cgttccggtg
     8941 ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc
     9001 cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc
     9061 agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc
     9121 ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc
     9181 gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac
     9241 ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt gctacgagat
     9301 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     9361 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     9421 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     9481 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     9541 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     9601 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     9661 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     9721 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     9781 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     9841 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     9901 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     9961 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
    10021 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
    10081 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
    10141 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
    10201 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
    10261 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
    10321 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
    10381 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
    10441 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
    10501 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
    10561 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
    10621 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
    10681 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
    10741 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
    10801 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
    10861 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
    10921 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
    10981 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
    11041 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
    11101 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
    11161 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
    11221 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
    11281 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
    11341 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
    11401 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
    11461 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
    11521 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
    11581 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
    11641 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
    11701 ggggttccgc gcacatttcc ccgaaaag
//
