LOCUS       sequence-181715-       12256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of S aureus SaCas9 (SauCas9) and two pol III promoters 
            for an Sa gRNA and an Sp gRNA. Intended to be used in 
            SpCas9-expressing cell lines for dual Cas9 ""Big Papi"" screens. aka
            pXPR207..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12256)
  AUTHORS   Najm FJ, Strand C, Donovan KF, Hegde M, Sanson KR, Vaimberg EW, 
            Sullender ME, Hartenian E, Kalani Z, Fusi N, Listgarten J, Younger 
            ST, Bernstein BE, Root DE, Doench JG
  TITLE     Orthologous CRISPR-Cas9 enzymes for combinatorial genetic screens.
  JOURNAL   Nat Biotechnol. 2018 Feb;36(2):179-189. doi: 10.1038/nbt.4048. Epub 
            2017 Dec 18.
  PUBMED    29251726
REFERENCE   2  (bases 1 to 12256)
  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/nbt.4048"; 
            journalName: "Nat Biotechnol"; date: "2018-02"; volume: "36"; issue:
            "2"; pages: "179-189"
COMMENT     Sequence Label: pPapi
FEATURES             Location/Qualifiers
     source          1..12256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(102..158)
                     /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             complement(168..215)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(216..3371)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Staphylococcus aureus 
                     Type II CRISPR/Cas system"
                     /label=SaCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="KRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEG
                     RRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEF
                     SAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEV
                     RGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGW
                     KDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQ
                     IIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEI
                     IENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAI
                     NLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIK
                     VINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAK
                     YLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQ
                     EENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFS
                     VQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERN
                     KGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIF
                     ITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKDN
                     DKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKD
                     NGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNL
                     DVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLLN
                     RIEVNMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQI
                     IKKG"
     CDS             complement(3396..3416)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      3419..3428
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3436..3647)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     promoter        3703..3917
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     3838..3857
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     promoter        complement(3959..4207)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(4017..4036)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(4187..4207)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    4258..4375
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(4673..4717)
                     /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    4902..5135
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    5628..5753
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             5800..5980
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(5998..6196)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6023..6043)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6198..6577
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     6749..6768
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6843..6947
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6948..7016,7017..7808)
                     /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     complement(7166..7185)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7979..8567
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8468..8487
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8721..8738
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8855..8876
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8891..8896,8897..8914,8915..8921)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8929..8945
                     /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)."
     primer_bind     8934..8956
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8953..8969
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8953..8969
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    9018..9139
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9029..9048)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     9083..9102
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(9269..9643)
                     /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"
     promoter        complement(9662..9709)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(9757..10086)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9771..9906
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9825..9844)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     10071..10091
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(10100..10528)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10210..10231)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10422..10441
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    10574..10798
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     10787..10804
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             10830..11063
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    11141..11729
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             11247..11258
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     11656..11676
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 gctgggtagt cggcgaaggc agcagccagc gtgcggacgg ccctgggcac atcgtcgcgg
       61 gttgccaagc ggacagttgg cttgtactcg gttccggagc cagggccagg gttctcctcg
      121 acgtcgccag cctgcttcag cagggaaaag ttggtggccc cggatccctt cttcttcttg
      181 gcctggccag ccttcttggt ggcagcaggg cgcttgccct tcttgatgat ctgagggtgc
      241 ttcttgctct tgacctcgta caagttgcca aggatgtcgg tggagtactt cttgatggac
      301 tgggtcttgg aggcgatggt cttgatgatg cgaggcgggc gcttgtcgtt catgttctcg
      361 aggtactcgc ggtaggtgat gtcgatcatg ttgacctcga tgcggttcag caggtcgttg
      421 ttgaccccga tgacgcggta gagctcgccg ttgatcttga tgaggtcgtt gttgtagaag
      481 ctggcgatga actcggcctg gttggagatc ttcttcaact ttttggcctc ctcgtagcac
      541 ttggagttga cctcgtagta gttctccttc ttgatgacgt ccaggttctt gacggtcacg
      601 aacttgtaga cgccgttgtc caggtagacg tcgaagcggt acggctttag cgacaacttt
      661 acaactttgt tgcgggagtt ggggtagtcg tcggtgatgt ccaggtgggc attcaacttg
      721 ttgccgtagt acttgatctt cttgatgacc gggccgttgt ccttcttgga gtacttggtg
      781 aggtagttgc cggtctcctc gtagtacttg tagagcgggt tcttctcgtc accgtactgc
      841 tccatgatca acttcaactt ttggtacgtc tgcgggtcat ggtggtacat gagcaacttt
      901 tcggggctct tgttgatcaa ctttttcaac ttgtcgttgt ccttgtcgta gaggccgttt
      961 aagttgttga cgatgagggt gttgcccttg tcgtccttgc gggtggagta gagggtgtcg
     1021 ttgatgagct cgcggttggg cttcttgtcc acgcggtggg agtacttgta gtccttgaag
     1081 tccttgatgt gcttgatctg gtgcggcgtg atgaagatct ccttgtactc ctgctctgtc
     1141 tcgatctccg gcatggactc ggcctgcttc tcctcgaaca tctggttctc catgaccttc
     1201 ttggccttgt ccaacttttt ccactccttg aagatgaagt cggcgttggc gatgatgagg
     1261 gcgtcctccg catggtgctt gtagcccttg ttgcgctcct tcttgaactt ccacttgcgg
     1321 cgcaggaagc tggtgaagcc accgttgatc gacttaactt tgacgtccaa gttgttgacc
     1381 cggaagtagc tgcgcagcag gttcatgagg ccacgggttg cgtagcgggt gtccaccagg
     1441 ttgcggttga tgaagtcctt ctgcaccgag aagcggttga tgtcgcgctc ctccagcagg
     1501 tactccttct tggtcttgga gatgcggccc ttgccctttg ccaagttgag gatgtgcttc
     1561 ttgaaagttt cgtaggagat cttggagtcc gaggaggaca ggtactggaa gggggtccgg
     1621 ttgcccttct tggagttctc ctcctgcttg accaggacct tgttgttgaa gctgttgtcg
     1681 aaggacaccg agcgcgggat gatgtggtca acttcgtagt tgaaggggtt gttgagcagg
     1741 tcctcgaggg ggatggcctc cagggagtag aggcacttgc cctcctgcat gtcatgcaac
     1801 tttatcttct cgatcaggta cttggcgttc tccttgccgg tggtgcggat gatctcctcg
     1861 atgcgctcgt tggtctggcg gttgcgcttc tgcatctcgt tgatcatctt ctgggcatcc
     1921 ttggagttct tctcgcgtgc caactcgatg atgatgtcgt tgggcaggcc gtacttcttg
     1981 atgatggcgt tgatgacctt gatggactgg atgaagctgc gcttgaccac gggcgagagg
     2041 atgaagtcgt cgaccagggt agttgggatc tccttctgtt gggacaggtc aactttcttg
     2101 ggcaccaact ttagtcggtt gaagatggcg atctggttgt cgttggtatg ccaaagttcg
     2161 tccaggatca agttgatggc cttgagggac aagttgtggg tgccagtgta gccctttaag
     2221 ttggagatct gctcgatctc ctcctgggta agctccgagt ttaagttggt cagctcctcc
     2281 tggatgtcct cggaactttg gtagatggtg aggatcttgg cgatctggtc cagcagctcc
     2341 gcgttctcga tgatctcctt gcgggcggtg atgtccttga tgtcatggta gacctttaag
     2401 ttggtgaact cgggcttgcc ggtggaggtg acccggtagc ccttgatgtc ctcctcgttg
     2461 accaggatct ccttggcgat ctgcttcaaa gttggcttct tcttctgctt gaagacgttt
     2521 tcgatgatct ggaacttttc gtagtactcc aacttttcgt tctcgtcgcg ggtgatgacc
     2581 aagttgttga ggtcgttgag cgcgttgtag aggtcggcgt tgtaggcgta cttgaccgag
     2641 cgcagctcct ccgggaagta ggtgcagtgg cccatgagca tctcgtacca ctccttgatg
     2701 tccttccagc cgaagggaga gccctcgcca ggcccctcgt agtaggtgcg gcgggtctcc
     2761 agcaggtcga tgtaggtgtc gatgaaggac tggtcaagtt ggtggtaggc cttctggacc
     2821 ttgagcagct gcttggcctc cttgacgtag tcggaggtct tgaagcggtt gatggagccg
     2881 cgcacctcgc cgtccttctt gaggcgctca agttggagct ccgcgacgta cttctcctcc
     2941 agggccttgg agttgcggga gatctgctcc ttggtggaaa gttcgttgcc ggtgtcctcc
     3001 tcaacttcgt tgacgttgtg gaccccgcgg cgcttggcca ggtgcagcag ggcagcagag
     3061 aactcctcct cggacaactt ttgcgagagg cccttgaccc gggcctcgta cgggttgatg
     3121 ccagacagct cggagtggtc ggtgagcaag ttgtagtcga agagcaactt tttcacgcgc
     3181 tggatgcggt ggcggcggcg gcgcttgagc cggcgggccc cgcgcttgga ccgccggccc
     3241 tcgttgttct cgacgttggc ctccttgaag aggcggacgc cagcgtcgat gacgtcccgg
     3301 gtctcgtagt cgatgatgcc atagccaact gaggtgatgc caatgtccag gcccaggatg
     3361 tagttgcgct tggcagcagg cacgccgtgg atgccaactt tgcgcttctt cttgggtgcc
     3421 atggtggcac cggtcctgtg ttctggcggc aaacccgttg cgaaaaagaa cgttcacggc
     3481 gactactgca cttatatacg gttctccccc accctcggga aaaaggcgga gccagtacac
     3541 gacatcactt tcccagttta ccccgcgcca ccttctctag gcaccggatc aattgccgac
     3601 ccctcccccc aacttctcgg ggactgtggg cgatgtgcgc tctgcccact gacgggcacc
     3661 ggagccaatt cccactcctt tcaagaccta gctagcgaat tcgaacgctg acgtcatcaa
     3721 cccgctccaa ggaatcgcgg gcccagtgtc actaggcggg aacacccagc gcgcgtgcgc
     3781 cctggcagga agatggctgt gagggacagg ggagtggcgc cctgcaatat ttgcatgtcg
     3841 ctatgtgttc tgggaaatca ccataaacgt gaaatgtctt tggatttggg aatcttataa
     3901 gttctgtatg agaccactct ttcccggaga cgtttttgcg cgcaaaaaac gcgtctccgg
     3961 tgtttcgtcc tttccacaag atatataaag ccaagaaatc gaaatacttt caagttacgg
     4021 taagcatatg atagtccatt ttaaaacata attttaaaac tgcaaactac ccaagaaatt
     4081 attactttct acgtcacgta ttttgtacta atatctttgt gtttacagtc aaattaattc
     4141 taattatctc tctaacagcc ttgtatcgta tatgcaaata tgaaggaatc atgggaaata
     4201 ggccctcggt accttaatta accaaactgg atctctgctg tccctgtaat aaacccgaaa
     4261 attttgaatt tttgtaattt gtttttgtaa ttctttagtt tgtatgtctg ttgctattat
     4321 gtctactatt ctttcccctg cactgtaccc cccaatcccc ccttttcttt taaaattgtg
     4381 gatgaatact gccatttgtc tgcagaattg gcgcacgcag tgccgatccg ttcactaatc
     4441 gaatggatct gtctctgtct ctctctccac cttcttcttc tattccttcg ggcctgtcgg
     4501 gtcccctcgg ggttgggagg tgggtctgaa acgataatgg tgaatatccc tgcctaactc
     4561 tattcactat agaaagtaca gcaaaaacta ttcttaaacc taccaagcct cctactatca
     4621 ttatgaataa ttttatatac cacagccaat ttgttatgtt aaaccaattc cacaaacttg
     4681 cccatttatc taattccaat aattcttgtt cattcttttc ttgctggttt tgcgattctt
     4741 caattaagga gtgtattaag cttgtgtaat tgttaatttc tctgtcccac tccatccagg
     4801 tcgtgtgatt ccaaatctgt tccagagatt tattactcca actagcattc caaggcacag
     4861 cagtggtgca aatgagtttt ccagagcaac cccaaatccc caggagctgt tgatccttta
     4921 ggtatctttc cacagccagg attcttgcct ggagctgctt gatgccccag actgtgagtt
     4981 gcaacagatg ctgttgcgcc tcaatagccc tcagcaaatt gttctgctgc tgcactatac
     5041 cagacaataa ttgtctggcc tgtaccgtca gcgtcattga cgctgcgccc atagtgcttc
     5101 ctgctgctcc caagaaccca aggaacaaag ctcctattcc cactgctctt ttttctctct
     5161 gcaccactct tctctttgcc ttggtgggtg ctactcctaa tggttcaatt tttactactt
     5221 tatatttata taattcactt ctccaattgt ccctcatatc tcctcctcca ggtctgaaga
     5281 tcagcggccg cttgctgtgc ggtggtctta cttttgtttt gctcttcctc tatcttgtct
     5341 aaagcttcct tggtgtcttt tatctctatc ctttgatgca cacaatagag ggttgctact
     5401 gtattatata atgatctaag ttcttctgat cctgtctgaa gggatggttg tagctgtccc
     5461 agtatttgtc tacagccttc tgatgtttct aacaggccag gattaactgc gaatcgttct
     5521 agctccctgc ttgcccatac tatatgtttt aatttatatt ttttctttcc ccctggcctt
     5581 aaccgaattt tttcccatcg cgatctaatt ctcccccgct taatactgac gctctcgcac
     5641 ccatctctct ccttctagcc tccgctagtc aaaatttttg gcgtactcac cagtcgccgc
     5701 ccctcgcctc ttgccgtgcg cgcttcagca agccgagtcc tgcgtcgaga gagctcctct
     5761 ggtttccctt tcgctttcaa gtccctgttc gggcgccact gctagagatt ttccacactg
     5821 actaaaaggg tctgagggat ctctagttac cagagtcaca caacagacgg gcacacacta
     5881 cttgaagcac tcaaggcaag ctttattgag gcttaagcag tgggttccct agttagccag
     5941 agagctccca ggctcagatc tggtctaacc agagagaccc agtacaggca aaacgcgctg
     6001 cttatataga cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg
     6061 ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca
     6121 atggggtgga gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac
     6181 tgccaaaacc gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta
     6241 ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg
     6301 acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt
     6361 taccgtaaat actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga
     6421 acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca
     6481 tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta
     6541 attgattact attaataact agtcaataat caatgtcaac gcgtatatct ggcccgtaca
     6601 tcgcgaagca gcgcaaaacg cctaacccta agcagattct tcatgcaatt gtcggtcaag
     6661 ccttgccttg ttgtagctta aattttgctc gcgcactact cagcgacctc caacacacaa
     6721 gcagggagca gatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
     6781 cataggggat cgggagatct cccgatccgt cgacgtcagg tggcactttt cggggaaatg
     6841 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6901 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     6961 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     7021 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     7081 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     7141 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     7201 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     7261 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     7321 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     7381 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     7441 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     7501 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     7561 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7621 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     7681 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7741 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7801 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     7861 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     7921 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     7981 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     8041 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     8101 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     8161 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     8221 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     8281 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     8341 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     8401 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     8461 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     8521 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     8581 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     8641 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     8701 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     8761 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     8821 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     8881 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     8941 aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag ctagaggtcg
     9001 acggtataca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     9061 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     9121 aagctgcaat aaacaagttg gggtgggcga agaactccag catgagatcc ccgcgctgga
     9181 ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg
     9241 cggtggaatc gaaatctcgt agcacgtgtc agtcctgctc ctcggccacg aagtgcacgc
     9301 agttgccggc cgggtcgcgc agggcgaact cccgccccca cggctgctcg ccgatctcgg
     9361 tcatggccgg cccggaggcg tcccggaagt tcgtggacac gacctccgac cactcggcgt
     9421 acagctcgtc caggccgcgc acccacaccc aggccagggt gttgtccggc accacctggt
     9481 cctggaccgc gctgatgaac agggtcacgt cgtcccggac cacaccggcg aagtcgtcct
     9541 ccacgaagtc ccgggagaac ccgagccggt cggtccagaa ctcgaccgct ccggcgacgt
     9601 cgcgcgcggt gagcaccgga acggcactgg tcaacttggc catggtttag ttcctcacct
     9661 tgtcgtatta tactatgccg atatactatg ccgatgatta attgtcaaca cgtgctgatc
     9721 agatccgaaa atggatatac aagctcccgg gagctttttg caaaagccta ggcctccaaa
     9781 aaagcctcct cactacttct ggaatagctc agaggcagag gcggcctcgg cctctgcata
     9841 aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg
     9901 cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat gcatgctttg
     9961 catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag
    10021 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg
    10081 acacacattc cacagaatta attcgcgtta aatttttgtt aaatcagctc attttttaac
    10141 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
    10201 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
    10261 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
    10321 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
    10381 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
    10441 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
    10501 gcgcttaatg cgccgctaca gggcgcgtgg ggataccccc tagagcccca gctggttctt
    10561 tccgcctcag aagccataga gcccaccgca tccccagcat gcctgctatt gtcttcccaa
    10621 tcctccccct tgctgtcctg ccccacccca ccccccagaa tagaatgaca cctactcaga
    10681 caatgcgatg caatttcctc attttattag gaaaggacag tgggagtggc accttccagg
    10741 gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga aggcacagtc
    10801 gaggctgatc agcgggttta aacgggccct gctagagatt ttccacactg actaaaaggg
    10861 tctgagggat ctctagttac cagagtcaca caacagacgg gcacacacta cttgaagcac
    10921 tcaaggcaag ctttattgag gcttaagcag tgggttccct agttagccag agagctccca
    10981 ggctcagatc tggtctaacc agagagaccc agtacaagca aaaagcagat cttgtcttcg
    11041 ttgggagtga attagccctt ccagtccccc cttttctttt aaaaagtggc taagatctac
    11101 agctgccttg taagtcattg gtcttaaagt cgacagcgct gcggggaggc ggcccaaagg
    11161 gagatccgac tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc
    11221 aggccgcggg aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc
    11281 ccgcgcagaa tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc
    11341 gacaacacca cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg
    11401 caggcggcga tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg
    11461 agctgacagg tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac
    11521 accacgccac gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat
    11581 ttatacaagg aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt
    11641 aaagcagcgt atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt
    11701 tcacaaattt tgtaatccag aggttgattg tcgacttaac gcgttctaga ttcatgcccc
    11761 aggcttgcgg gtcatgcacc aggtgcgcgg cccctcgggc acctcgacgt ccgcggtgac
    11821 ggtgaagccc aggcgctcgt agaagggcaa gttgcgaggg gcagaggtct ccaggaaggc
    11881 agggacgcca gcgcgctcgg cagcctcgac cccaggcagc accacggcgc tgcccaggcc
    11941 cttgccctgg tggtcgggag agacaccaac tgtggccagg aaccaggccg gctccttggg
    12001 gcggtggggt gccaacagcc cctccatctg ctgttgggca gccaaccggg agcccgaaag
    12061 ctcggccatg cgcggcccga tctcggcgaa gacggcgcca gcctcgaccg actcgggggt
    12121 ggtccagacg gccacggcag caccgtcgtc cgccacccag accttgccaa tgtcaaggcc
    12181 aactcgggtg aggaagagct cctgcagctc ggtgacgcgc tcgatgtggc ggtccgggtc
    12241 cacggtgtgg cgggtg
//
