LOCUS       sequence-363237-       12704 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for CRISPRi, lentiviral expression of KRAB-dCas9 and BlastR. Also 
            called pXPR_121.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12704)
  AUTHORS   Rosenbluh J, Xu H, Harrington W, Gill S, Wang X, Vazquez F, Root DE,
            Tsherniak A, Hahn WC
  TITLE     Complementary information derived from CRISPR Cas9 mediated gene 
            deletion and suppression.
  JOURNAL   Nat Commun. 2017 May 23;8:15403. doi: 10.1038/ncomms15403.
  PUBMED    28534478
REFERENCE   2  (bases 1 to 12704)
  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: "Nat 
            Commun."; date: "2017-05-23"; volume: "8:15403. doi"; pages: " 
            10.1038/ncomms15403"
COMMENT     Sequence Label: pLX_311-KRAB-dCas9
FEATURES             Location/Qualifiers
     source          1..12704
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             46..90
                     /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"
     promoter        321..531
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        343..539
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      390..525
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             561..956
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    1030..1147
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1285..2463
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1516..2454
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2411..2431
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    2483..2507
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2537..2722
                     /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             join(2744..2770,2771..2773,2774..5260,5261..5263,
                     5264..6847)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             2744..6847
                     /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
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6851..6877
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6857..6883
                     /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             6896..6916
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6923..6943
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(6988..7012)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             7020..7061
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    7112..7700
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7165..7185)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7583..7594)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7772..8005
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8083..8204
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8120..8139)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8174..8193
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8244..8379
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8399..8418)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8400..8418)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8428..8445)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8428..8444)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8437..8459)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8586..9041
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8673..8692)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8883..8904
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9067..9171
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9172..9240,9241..10032)
                     /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(9390..9409)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10203..10791
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10692..10711
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10945..10962
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11079..11100
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11115..11120,11121..11138,11139..11145)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11153..11169
                     /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     11158..11180
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11177..11193
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11177..11193
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     11212..11232
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        11214..11232
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        11260..11486
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             11487..11667
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11714..11839
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    12332..12565
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa
       61 ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg
      121 tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt
      181 gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc
      241 cacctcccaa ccccgagggg acccatgcat tgcatctcaa ttagtcagca accaggtgtg
      301 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcgtctc aattagtcag
      361 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      421 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
      481 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      541 agctttctag aggtaccacc atggccaagc ctttgtctca agaagaatcc accctcattg
      601 aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc gtcgccagcg
      661 cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat tttactgggg
      721 gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct ggcaacctga
      781 cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc ggacggtgcc
      841 gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac agtgatggac
      901 agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg gagggcctgc
      961 agctgcagta gtaaggcgcg ccgttaacga attctagatc ttgagacaaa tggcagtatt
     1021 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     1081 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     1141 aaattttcgg gtttattaca gggacagcag agatccactt tggcgccggc tcgaggcctg
     1201 caggtgcaaa gatggataaa gttttaaaca gagaggaatc tttgcagcta atggaccttc
     1261 taggtcttga aaggagtggg aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg
     1321 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg
     1381 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     1441 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     1501 gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt
     1561 tatggccctt gcgtgccttg aattacttcc acctggctgc agtacgtgat tcttgatccc
     1621 gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt gcgcttaagg agccccttcg
     1681 cctcgtgctt gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg
     1741 caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa tttttgatga
     1801 cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca agatctgcac
     1861 actggtattt cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca
     1921 tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa
     1981 gctggccggc ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg
     2041 gcaaggctgg cccggtcggc accagttgcg tgagcggaaa gatggccgct tcccggccct
     2101 gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc
     2161 acacaaagga aaagggcctt tccgtcctca gccgtcgctt catgtgactc cacggagtac
     2221 cgggcgccgt ccaggcacct cgattagttc tcgagctttt ggagtacgtc gtctttaggt
     2281 tggggggagg ggttttatgc gatggagttt ccccacactg agtgggtgga gactgaagtt
     2341 aggccagctt ggcacttgat gtaattctcc ttggaatttg ccctttttga gtttggatct
     2401 tggttcattc tcaagcctca gacagtggtt caaagttttt ttcttccatt tcaggtgtcg
     2461 tgaggctagc atcgattgat caacaagttt gtacaaaaaa gttggcatgg atgctaagtc
     2521 actaactgcc tggtcccgga cactggtgac cttcaaggat gtatttgtgg acttcaccag
     2581 ggaggagtgg aagctgctgg acactgctca gcagatcgtg tacagaaatg tgatgctgga
     2641 gaactataag aacctggttt ccttgggtta tcagcttact aagccagatg tgatcctccg
     2701 gttggagaag ggagaagagc ccggcggttc cggcggaggg tcgatggaca agaagtacag
     2761 catcggcctg gccatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa
     2821 ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa
     2881 cctgatcggc gccctgctgt tcgacagcgg agaaacagcc gaggccaccc ggctgaagag
     2941 aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt
     3001 cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag agtccttcct
     3061 ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt
     3121 ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac
     3181 cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg
     3241 ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat
     3301 ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt
     3361 ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat
     3421 cgcccagctg cccggcgaga agaagaatgg cctgttcggc aacctgattg ccctgagcct
     3481 gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct
     3541 gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta
     3601 cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct
     3661 gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga
     3721 cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa
     3781 gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca tcgatggcgg
     3841 agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac
     3901 cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga
     3961 caacggcagc atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca
     4021 ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt
     4081 ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac
     4141 cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc
     4201 cagcgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa
     4261 ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtacaacg agctgaccaa
     4321 agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa
     4381 agccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga
     4441 ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg
     4501 gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt
     4561 cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt
     4621 tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa
     4681 agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct
     4741 gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga
     4801 cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga
     4861 ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa
     4921 tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga
     4981 gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga
     5041 gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga
     5101 gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct
     5161 gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca
     5221 ggaactggac atcaaccggc tgtccgacta cgatgtggac gctatcgtgc ctcagagctt
     5281 tctgaaggac gactccatcg ataacaaagt gctgactcgg agcgacaaga accggggcaa
     5341 gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcgccagct
     5401 gctgaatgcc aagctgatta cccagaggaa gttcgacaat ctgaccaagg ccgagagagg
     5461 cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca
     5521 gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa
     5581 cgacaaactg atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt
     5641 ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga
     5701 cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag
     5761 cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga
     5821 gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt
     5881 caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa
     5941 cggcgaaaca ggcgagatcg tgtgggataa gggccgggac tttgccaccg tgcggaaagt
     6001 gctgtctatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag
     6061 caaagagtct atcctgccca agaggaacag cgacaagctg atcgccagaa agaaggactg
     6121 ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt
     6181 ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat
     6241 caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg
     6301 ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct
     6361 ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact
     6421 ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa
     6481 gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca aacactacct
     6541 ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa
     6601 tctggacaag gtgctgagcg cctacaacaa gcacagagac aagcctatca gagagcaggc
     6661 cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta
     6721 ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac
     6781 cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg
     6841 aggcgacgcc tatccctatg acgtgcccga ttatgccagc ctgggcagcg gctcccccaa
     6901 gaaaaaacgc aaggtggaag atcctaagaa aaagcggaaa gtggacggca ttggtagtgg
     6961 gagcaacggc agcagcggat cctaaacccc caactttctt gtacaaagtg gttgatatcg
     7021 gtaagcctat ccctaaccct ctcctcggtc tcgattctac gtagtaatga actagtaccg
     7081 gttaagtcga caatcaacgc gttaagtcga caatcaacct ctggattaca aaatttgtga
     7141 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     7201 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     7261 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     7321 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     7381 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     7441 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     7501 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     7561 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     7621 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     7681 cctttgggcc gcctccccgc gtcgacttta agaccaatga cttacaaggc agctgtagat
     7741 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     7801 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     7861 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     7921 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     7981 tagtcagtgt ggaaaatctc tagcagtacg tatagtagtt catgtcatct tattattcag
     8041 tatttataac ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct
     8101 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     8161 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag
     8221 ctatcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     8281 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     8341 agctattcca gaagtagtga ggaggctttt ttggaggcct agggacgtac ccaattcgcc
     8401 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     8461 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     8521 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     8581 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     8641 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     8701 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     8761 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     8821 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     8881 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     8941 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     9001 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     9061 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     9121 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     9181 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     9241 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     9301 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     9361 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     9421 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     9481 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     9541 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     9601 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     9661 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     9721 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     9781 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     9841 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     9901 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     9961 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
    10021 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
    10081 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
    10141 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
    10201 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
    10261 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
    10321 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
    10381 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
    10441 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
    10501 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
    10561 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
    10621 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
    10681 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
    10741 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
    10801 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
    10861 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
    10921 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
    10981 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
    11041 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
    11101 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
    11161 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
    11221 cctcactaaa gggaacaaaa gctggagctg caagcttaat gtagtcttat gcaatactct
    11281 tgtagtcttg caacatggta acgatgagtt agcaacatgc cttacaagga gagaaaaagc
    11341 accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac
    11401 agacgggtct gacatggatt ggacgaacca ctgaattgcc gcattgcaga gatattgtat
    11461 ttaagtgcct agctcgatac ataaacgggt ctctctggtt agaccagatc tgagcctggg
    11521 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
    11581 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
    11641 tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga aagcgaaagg
    11701 gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca cggcaagagg
    11761 cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct agaaggagag
    11821 agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat gggaaaaaat
    11881 tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat gggcaagcag
    11941 ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag gctgtagaca
    12001 aatactggga cagctacaac catcccttca gacaggatca gaagaactta gatcattata
    12061 taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag acaccaagga
    12121 agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac agcaagcggc
    12181 cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt gaattatata
    12241 aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca aagagaagag
    12301 tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg ttcttgggag
    12361 cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc agacaattat
    12421 tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg caacagcatc
    12481 tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg gctgtggaaa
    12541 gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa ctcatttgca
    12601 ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag atttggaatc
    12661 acacgacctg gatggagtgg gacagagaaa ttaacaatta caca
//
