LOCUS       sequence-141793-       11386 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Cas9 + sgGFP-1 with blasticidin selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11386)
  AUTHORS   Wang B, Krall EB, Aguirre AJ, Kim M, Widlund HR, Doshi MB, Sicinska 
            E, Sulahian R, Goodale A, Cowley GS, Piccioni F, Doench JG, Root DE,
            Hahn WC
  TITLE     ATXN1L, CIC, and ETS Transcription Factors Modulate Sensitivity to 
            MAPK Pathway Inhibition.
  JOURNAL   Cell Rep. 2017 Feb 7;18(6):1543-1557. doi: 
            10.1016/j.celrep.2017.01.031.
  PUBMED    28178529
REFERENCE   2  (bases 1 to 11386)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.celrep.2017.01"; journalName: "Cell Rep"; date: 
            "2017-02-7- 7"; volume: "18"; issue: "6"; pages: "1543-1557"
COMMENT     Sequence Label: PXPR007 sgGFP-1
FEATURES             Location/Qualifiers
     source          1..11386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        1..76
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    124..241
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        310..521
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      541..550
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             550..573
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             580..600
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             625..4725
                     /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             625..4725
                     /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             4726..4773
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4789..4845
                     /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             4846..5241
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    5257..5845
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5310..5330)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5728..5739)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5917..6150
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6228..6349
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6265..6284)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6319..6338
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6389..6524
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6544..6563)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6545..6563)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6573..6590)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6573..6589)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6582..6604)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6731..7186
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6818..6837)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7028..7049
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7212..7316
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7317..7385,7386..8177)
                     /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(7535..7554)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8348..8936
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8837..8856
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9090..9107
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9224..9245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9260..9265,9266..9283,9284..9290)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9298..9314
                     /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     9303..9325
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9322..9338
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9322..9338
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9357..9377
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9359..9377
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9405..9631
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             9632..9812
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9859..9984
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10477..10710
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10895..10939
                     /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        11117..11365
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     11117..11137
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     11288..11307
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
       61 ggcaccgagt cggtgctttt ttgaattcta gatcttgaga caaatggcag tattcatcca
      121 caattttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
      181 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
      241 tcgggtttat tacagggaca gcagagatcc actttggcgc cggctcgagt ggctccggtg
      301 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      361 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      421 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      481 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtgtcgtga cgcgggatcc
      541 gccaccatgg attacaaaga cgatgacgat aagatggccc caaagaagaa gcggaaggtc
      601 ggtatccacg gagtcccagc agccgacaag aagtacagca tcggcctgga catcggcacc
      661 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
      721 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc
      781 gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc
      841 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
      901 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
      961 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
     1021 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
     1081 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
     1141 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
     1201 cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct
     1261 gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag
     1321 aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
     1381 agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac
     1441 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
     1501 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
     1561 aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc
     1621 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
     1681 cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga agagttctac
     1741 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
     1801 aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag
     1861 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
     1921 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
     1981 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
     2041 accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag
     2101 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
     2161 ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga
     2221 atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc
     2281 aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag
     2341 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
     2401 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
     2461 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
     2521 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
     2581 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
     2641 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
     2701 atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg
     2761 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
     2821 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg
     2881 cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga
     2941 cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc
     3001 cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg
     3061 tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg
     3121 tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac
     3181 aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa
     3241 gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc
     3301 cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag
     3361 gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag
     3421 atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg
     3481 aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac
     3541 aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg
     3601 ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac
     3661 aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc
     3721 gccaagtact tcttctacag caacatcatg aactttttca agaccgagat taccctggcc
     3781 aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg
     3841 tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat
     3901 atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag
     3961 aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc
     4021 ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag
     4081 tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc
     4141 ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac
     4201 ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg
     4261 ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg
     4321 aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag
     4381 cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc
     4441 agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc
     4501 tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt
     4561 accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg
     4621 aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
     4681 ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacaagcg tcctgctgct
     4741 actaagaaag ctggtcaagc taagaaaaag aaagctagcg gcagcggcgc caccaacttc
     4801 agcctgctga agcaggccgg cgacgtggag gagaaccccg gccccatggc caagcctttg
     4861 tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc
     4921 tctgaagact acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt
     4981 gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct
     5041 gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc
     5101 atcttgagcc cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa
     5161 gccatagtga aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc
     5221 tctggttatg tgtgggaggg cacgcgttaa gtcgacaatc aacctctgga ttacaaaatt
     5281 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     5341 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     5401 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     5461 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     5521 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     5581 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     5641 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     5701 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     5761 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     5821 atctcccttt gggccgcctc cccgcgtcga ctttaagacc aatgacttac aaggcagctg
     5881 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     5941 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
     6001 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     6061 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     6121 ccttttagtc agtgtggaaa atctctagca gtacgtatag tagttcatgt catcttatta
     6181 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     6241 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6301 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     6361 tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     6421 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     6481 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggga cgtacccaat
     6541 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac
     6601 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     6661 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     6721 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     6781 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     6841 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     6901 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     6961 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     7021 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     7081 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     7141 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     7201 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     7261 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     7321 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     7381 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     7441 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     7501 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     7561 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     7621 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     7681 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     7741 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     7801 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     7861 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     7921 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     7981 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     8041 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     8101 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     8161 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     8221 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     8281 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     8341 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     8401 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     8461 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     8521 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     8581 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     8641 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     8701 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     8761 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     8821 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     8881 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     8941 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     9001 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     9061 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     9121 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     9181 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     9241 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     9301 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa
     9361 ttaaccctca ctaaagggaa caaaagctgg agctgcaagc ttaatgtagt cttatgcaat
     9421 actcttgtag tcttgcaaca tggtaacgat gagttagcaa catgccttac aaggagagaa
     9481 aaagcaccgt gcatgccgat tggtggaagt aaggtggtac gatcgtgcct tattaggaag
     9541 gcaacagacg ggtctgacat ggattggacg aaccactgaa ttgccgcatt gcagagatat
     9601 tgtatttaag tgcctagctc gatacataaa cgggtctctc tggttagacc agatctgagc
     9661 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     9721 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     9781 acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga cttgaaagcg
     9841 aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc gcgcacggca
     9901 agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg aggctagaag
     9961 gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc gcgatgggaa
    10021 aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat agtatgggca
    10081 agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc agaaggctgt
    10141 agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca
    10201 ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat aaaagacacc
    10261 aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac cgcacagcaa
    10321 gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt
    10381 atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag
    10441 aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt
    10501 gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca
    10561 attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca
    10621 gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt
    10681 ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat
    10741 ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg
    10801 gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca
    10861 ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt
    10921 agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa
    10981 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
    11041 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
    11101 cccgagggga cccagagagg gcctatttcc catgattcct tcatatttgc atatacgata
    11161 caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca
    11221 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
    11281 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
    11341 tatcttgtgg aaaggacgaa acaccgggcg agggcgatgc caccta
//
