LOCUS       sequence-261405-       12420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EF1a promoter expresses SpyoCas9 and P2A provides EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12420)
  AUTHORS   DeWeirdt PC, Sangree AK, Hanna RE, Sanson KR, Hegde M, Strand C, 
            Persky NS, Doench JG
  TITLE     Genetic screens in isogenic mammalian cell lines without single cell
            cloning.
  JOURNAL   Nat Commun. 2020 Feb 6;11(1):752. doi: 10.1038/s41467-020-14620-6.
  PUBMED    32029722
REFERENCE   2  (bases 1 to 12420)
  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: "2020-02-6"; volume: "11(1):752. doi"; pages: " 
            10.1038/s41467-020-14620-6"
COMMENT     Sequence Label: pXPR_124
FEATURES             Location/Qualifiers
     source          1..12420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..181
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    228..353
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    846..1079
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1264..1308
                     /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    1574..1691
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    1709..1729
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        1730..2908
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1961..2899
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2856..2876
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    2908..2932
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2933..2942
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2942..3007
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3014..3034
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3059..7159
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             3059..7159
                     /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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7160..7207
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7217..7273
                     /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             join(7274..7276,7277..7279,7280..7993)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7274..7276,7277..7279,7280..7990)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7580..7599)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7927..7948
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(7994..8018)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    8036..8624
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8089..8109)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8507..8518)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8712..8945
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9017..9138
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9054..9073)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9108..9127
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9178..9313
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9333..9352)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9334..9352)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9362..9379)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9362..9378)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9371..9393)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9520..9975
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9607..9626)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9817..9838
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10001..10105
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10106..10174,10175..10966)
                     /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(10324..10343)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11137..11725
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11626..11645
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11879..11896
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12013..12034
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12049..12054,12055..12072,12073..12079)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12087..12103
                     /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     12092..12114
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     12111..12127
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12111..12127
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     12146..12166
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        12148..12166
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        12194..12420
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc
      661 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg
     1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt
     1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga
     1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa
     1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt
     1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta
     1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca
     1441 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata
     1501 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga
     1561 cggtatcgct agcttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa
     1621 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa
     1681 ttcaaaattt tactagtgat tatcggatca actttgtata gaaaagttgg gctccggtgc
     1741 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
     1801 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
     1861 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
     1921 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc
     1981 ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg
     2041 gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag
     2101 gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg
     2161 gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc
     2221 tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct
     2281 tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga
     2341 cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc
     2401 gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gtctcgcgcc
     2461 gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc
     2521 ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc
     2581 gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt
     2641 cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag
     2701 cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca
     2761 cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga
     2821 atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag
     2881 tttttttctt ccatttcagg tgtcgtgaca agtttgtaca aaaaagcagg ctgccaccat
     2941 ggactataag gaccacgacg gagactacaa ggatcatgat attgattaca aagacgatga
     3001 cgataagatg gccccaaaga agaagcggaa ggtcggtatc cacggagtcc cagcagccga
     3061 caagaagtac agcatcggcc tggacatcgg caccaactct gtgggctggg ccgtgatcac
     3121 cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag
     3181 catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac
     3241 ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct
     3301 gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga
     3361 agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat
     3421 cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact
     3481 ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat
     3541 caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga
     3601 caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa
     3661 cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct
     3721 ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat
     3781 tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc
     3841 caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat
     3901 cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct
     3961 gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat
     4021 caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca
     4081 gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta
     4141 cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa
     4201 gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca
     4261 gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat
     4321 tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa
     4381 gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt
     4441 cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt
     4501 ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct
     4561 gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa
     4621 cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg
     4681 cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa
     4741 gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg
     4801 cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa
     4861 ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac
     4921 cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct
     4981 gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct
     5041 gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt
     5101 cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct
     5161 gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga
     5221 gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa
     5281 ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga
     5341 aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa
     5401 gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga
     5461 aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat
     5521 gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt
     5581 gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa
     5641 gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta
     5701 ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa
     5761 ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt
     5821 ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa
     5881 gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct
     5941 ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca
     6001 ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc
     6061 taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat
     6121 cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat
     6181 catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct
     6241 gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac
     6301 cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac
     6361 aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag
     6421 aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc
     6481 tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga
     6541 gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct
     6601 ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc
     6661 cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa
     6721 gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta
     6781 tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca
     6841 caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct
     6901 ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat
     6961 cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc
     7021 cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt
     7081 gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct
     7141 gtctcagctg ggaggcgaca aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa
     7201 gaaaaagggc agcggcgcca cgaacttctc tctgttaaag caagcaggag atgttgaaga
     7261 aaaccccggg cctatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     7321 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     7381 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     7441 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     7501 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     7561 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     7621 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     7681 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     7741 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     7801 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     7861 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     7921 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     7981 gctgtacaag taaacccagc tttcttgtac aaagtggtga taatcgaatt ccgataatca
     8041 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     8101 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     8161 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     8221 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     8281 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     8341 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     8401 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     8461 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     8521 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     8581 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcggg aattcccgcg
     8641 gttcgcttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     8701 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
     8761 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     8821 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     8881 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     8941 tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat
     9001 atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     9061 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     9121 actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccatc
     9181 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     9241 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     9301 ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct
     9361 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     9421 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     9481 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     9541 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     9601 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     9661 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     9721 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     9781 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     9841 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     9901 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     9961 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
    10021 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
    10081 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
    10141 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
    10201 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
    10261 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
    10321 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
    10381 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
    10441 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
    10501 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
    10561 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
    10621 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
    10681 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
    10741 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
    10801 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
    10861 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
    10921 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
    10981 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
    11041 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
    11101 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
    11161 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
    11221 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
    11281 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
    11341 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
    11401 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
    11461 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
    11521 gcgtgagcta tgagaaagcg ccacgcttcc cgaagagaga aaggcggaca ggtatccggt
    11581 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
    11641 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
    11701 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
    11761 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
    11821 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
    11881 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
    11941 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
    12001 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
    12061 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
    12121 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga
    12181 gctgcaagct taatgtagtc ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg
    12241 agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt ggtggaagta
    12301 aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg gattggacga
    12361 accactgaat tgccgcattg cagagatatt gtatttaagt gcctagctcg atacataaac
//
