LOCUS       sequence-239465-       14975 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Catalytically inactive Cas9 from S. pyogenes with P2A-BlastR under 
            the EF1a core promoter, and cloning backbone for sgRNA. Contains 
            BsmBI sites for insertion of spacer sequences..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14975)
  TITLE     Ferrer Lab Plasmids
REFERENCE   2  (bases 1 to 14975)
  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"
COMMENT     Sequence Label: Lenti-(BB)-EF1a-KRAB-dCas9-P2A-BlastR
FEATURES             Location/Qualifiers
     source          1..14975
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    24..612
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(77..97)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(495..506)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             684..917
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(943..960)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    949..1173
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1219..1647
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1306..1325)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1516..1537
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1656..1676)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1661..1990
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1841..1976
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1903..1922
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2038..2085
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2104..2478
                     /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    2608..2729
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2645..2664)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2699..2718
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2778..2794)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2778..2794)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2791..2813)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2802..2818
                     /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(2826..2832,2833..2850,2851..2856))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2871..2892
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3009..3026)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3180..3768)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3260..3279)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3939..4730,4731..4799))
                     /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     4562..4581
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4800..4904)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4979..4998)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5170..5549
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5551..5749
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5704..5724
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5767..5947
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5994..6119
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6612..6845
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7030..7074
                     /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    7372..7489
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7540..7788
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7540..7560
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7711..7730
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9670..9745
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        9807..10018
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      10031..10040
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             10067..10252
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     CDS             10274..14377
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTSLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             14381..14407
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             14387..14413
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             14426..14446
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             14453..14473
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             14519..14575
                     /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             14576..14974
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
ORIGIN
        1 aattcgatat caagcttatc ggtaatcaac ctctggatta caaaatttgt gaaagattga
       61 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
      121 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
      181 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
      241 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
      301 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
      361 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
      421 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
      481 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
      541 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
      601 ccgcctcccc gcgtcgactt taagaccaat gacttacaag gcagctgtag atcttagcca
      661 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct
      721 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
      781 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
      841 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
      901 gtggaaaatc tctagcaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta
      961 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     1021 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     1081 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     1141 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     1201 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     1261 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     1321 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     1381 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     1441 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     1501 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     1561 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     1621 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     1681 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     1741 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     1801 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     1861 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     1921 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     1981 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt
     2041 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa
     2101 accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg
     2161 gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc
     2221 ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg
     2281 gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg
     2341 gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag
     2401 cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg
     2461 gccgaggagc aggactgaca cgtgctacga gatttcgatt ccaccgccgc cttctatgaa
     2521 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     2581 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     2641 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2701 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     2761 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     2821 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     2881 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2941 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3001 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3061 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3121 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3181 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3241 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3301 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3361 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3421 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3481 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3541 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3601 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3661 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3721 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3781 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3841 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3901 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3961 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4021 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4081 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4141 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4201 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4261 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4321 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4381 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4441 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4501 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4561 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4621 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4681 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     4741 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     4801 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     4861 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     4921 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
     4981 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     5041 tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt
     5101 gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat
     5161 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     5221 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     5281 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     5341 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     5401 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     5461 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     5521 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     5581 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     5641 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     5701 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcagc gcgttttgcc
     5761 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     5821 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     5881 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     5941 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
     6001 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     6061 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     6121 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     6181 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     6241 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     6301 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     6361 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     6421 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga
     6481 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
     6541 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
     6601 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
     6661 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
     6721 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
     6781 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
     6841 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
     6901 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg
     6961 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac
     7021 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat
     7081 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
     7141 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
     7201 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa
     7261 ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga
     7321 tcggcactgc gtgcgccaat tctgcagaca aatggcagta ttcatccaca attttaaaag
     7381 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     7441 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta
     7501 cagggacagc agagatccag tttggttaat taaggtaccg agggcctatt tcccatgatt
     7561 ccttcatatt tgcatatacg atacaaggct gttagagaga taattagaat taatttgact
     7621 gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag
     7681 tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag
     7741 tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgg agacggttgt
     7801 aaatgagcac acaaaataca catgctaaaa tattatattc tatgaccttt ataaaatcaa
     7861 ccaaaatctt ctttttaata actttagtat caataattag aatttttatg ttcctttttg
     7921 caaactttta ataaaaatga gcaaaataaa aaaacgctag ttttagtaac tcgcgttgtt
     7981 ttcttcacct ttaataatag ctactccacc acttgttcct aagcggtcag ctcctgcttc
     8041 aatcattttt tgagcatctt caaatgttct aactccacca gctgctttaa ctaaagcatt
     8101 gtctttaaca actgacttca ttagtttaac atcttcaaat gttgcacctg attttgaaaa
     8161 tcctgttgat gttttaacaa attctaatcc agcttcaaca gctatttcac aagctttcat
     8221 gatttcttct tttgttaata aacaattttc cataatacat ttaacaacat gtgatccagc
     8281 tgcttttttt acagctttca tgtcttctaa aactaattca taatttttgt cttttaatgc
     8341 accaatattt aataccatat caatttctgt tgcaccatct ttaattgctt cagaaacttc
     8401 gaatgctttt gtagctgttg tgcatgcacc tagaggaaaa cctacaacat ttgttattcc
     8461 tacatttgtg ccttttaata attctttaca atagcttgtt caatatgaat taacacaaac
     8521 tgttgcaaaa tcaaattcaa ttgcttcatc acataattgt ttaatttcag ctttcgtagc
     8581 atcttgtttt aataatgtgt gatctatata tttgtttagt ttcatttttt ctcctatata
     8641 ttcattttta attttaattc tttaataatt tcgtctactt taactttagc gttttgaaca
     8701 gattcaccaa cacctataaa ataaattttt agtttaggtt cagttccact tgggcgaaca
     8761 gcaaatcatg acttatcttc taaataaaat tttagtaagt cttgtcctgg catattatac
     8821 attccatcga tgtagtcttc aacattaaca actttaagtc cagcaatttg agttaagggt
     8881 gttgctctca atgatttcat taatggttca atttttaatt tcttttcttc tggtttaaaa
     8941 ttcaagttta aagtgaaagt gtaatatgca cccatttctt taaataaatc ttctaaatag
     9001 tctactaatg ttttattttg ttttttataa aatcaagcag cctctgctat taatatagaa
     9061 gcttgtattc catctttatc tctagctgag tcatcaatta catatccata actttcttca
     9121 taagcaaaaa caaaatttaa tccgttatct tcttctttag caatttctct acccattcat
     9181 ttaaatccag ttaaagtttt tacaatatta actccatatt tttcatgagc gattctatca
     9241 cccaaatcac ttgttacaaa acttgaatat agagccggat tttttggaat gctatttaag
     9301 cgttttagat ttgataattt tcaatcaatt aaaattggtc ctgtttgatt tccatctaat
     9361 cttacaaaat gaccatcatg ttttattgcc attccaaatc tgtcagcatc tgggtcattc
     9421 ataataataa tatctgcatc atgtttaata ccatattcaa gcggtatttt tcatgcagga
     9481 tcaaattctg gatttggatt tacaacattt ttaaatgttt catcttcaaa tgcatgctct
     9541 tcaacctcaa taacgttata tcctgattca cgtaatattt ttggggtaaa tttagttcct
     9601 gttccattaa ctgcgctaaa aataattttt aaatcttttt tagcttcttg ctcttttttg
     9661 tacgtctctg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
     9721 tgaaaaagtg gcaccgagtc ggtgcttttt tgaattcgct agctaggtct tgaaaggagt
     9781 gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa
     9841 gttgggggga ggggtcggca attgatccgg tgcctagaga aggtggcgcg gggtaaactg
     9901 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat
     9961 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagga
    10021 ccggtgatcc gccaccatgg atgctaagtc actaactgcc tggtcccgga cactggtgac
    10081 cttcaaggat gtatttgtgg acttcaccag ggaggagtgg aagctgctgg acactgctca
    10141 gcagatcgtg tacagaaatg tgatgctgga gaactataag aacctggttt ccttgggtta
    10201 tcagcttact aagccagatg tgatcctccg gttggagaag ggagaagagc ccggcggttc
    10261 cggcggaggg tcgatggaca agaagtacag catcggcctg gccatcggca ccaactctgt
    10321 gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg
    10381 caacaccgac cggcacagca tcaagaagaa cctgatcggc gccctgctgt tcgacagcgg
    10441 agaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa
    10501 gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag
    10561 cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca
    10621 ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta
    10681 ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct
    10741 ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc
    10801 cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt
    10861 cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact
    10921 gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg
    10981 cctgttcggc aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt
    11041 cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga
    11101 caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct
    11161 gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc
    11221 cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa
    11281 agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa
    11341 gaacggctac gccggctaca tcgatggcgg agccagccag gaagagttct acaagttcat
    11401 caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga
    11461 ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct
    11521 gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa
    11581 ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc
    11641 caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg
    11701 gaacttcgag gaagtggtgg acaagggcgc cagcgcccag agcttcatcg agcggatgac
    11761 caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga
    11821 gtacttcacc gtgtacaacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa
    11881 gcccgccttc ctgagcggcg agcagaaaaa agccatcgtg gacctgctgt tcaagaccaa
    11941 ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga
    12001 ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga
    12061 tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct
    12121 ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct
    12181 gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata
    12241 caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg
    12301 caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct
    12361 gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca
    12421 gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg
    12481 catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc
    12541 cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa
    12601 cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct
    12661 gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct
    12721 gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta
    12781 cgatgtggac gctatcgtgc ctcagagctt tctgaaggac gactccatcg ataacaaagt
    12841 gctgactcgg agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt
    12901 gaagaagatg aagaactact ggcgccagct gctgaatgcc aagctgatta cccagaggaa
    12961 gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt
    13021 catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga
    13081 ctcccggatg aacactaagt acgacgagaa cgacaaactg atccgggaag tgaaagtgat
    13141 caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg
    13201 cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc
    13261 cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta
    13321 cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta
    13381 cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga
    13441 gatccggaag cggcctctga tcgagacaaa cggcgaaaca ggcgagatcg tgtgggataa
    13501 gggccgggac tttgccaccg tgcggaaagt gctgtctatg ccccaagtga atatcgtgaa
    13561 aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag
    13621 cgacaagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag
    13681 ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa
    13741 actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa
    13801 gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat
    13861 caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc
    13921 tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct
    13981 gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca
    14041 gctgtttgtg gaacagcaca aacactacct ggacgagatc atcgagcaga tcagcgagtt
    14101 ctccaagaga gtgatcctgg ccgacgctaa tctggacaag gtgctgagcg cctacaacaa
    14161 gcacagagac aagcctatca gagagcaggc cgagaatatc atccacctgt ttaccctgac
    14221 cagtctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta
    14281 caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta
    14341 cgagacacgg atcgacctgt ctcagctggg aggcgacgcc tatccctatg acgtgcccga
    14401 ttatgccagc ctgggcagcg gctcccccaa gaaaaaacgc aaggtggaag atcctaagaa
    14461 aaagcggaaa gtggacggca ttggtagtgg gagcaacggc agcagcgggg atccggccgc
    14521 aacaaacttc tctctgctga aacaagccgg agatgtcgaa gagaatcctg gaccgatggc
    14581 caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag
    14641 catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg acggccgcat
    14701 cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct
    14761 gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga
    14821 gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg atctgcatcc
    14881 tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg ggattcgtga
    14941 attgctgccc tctggttatg tgtgggaggg ctaag
//
