LOCUS       sequence-132101-        9293 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes gRNA for 3' target of human ATF4 along with Cas9 with 2A 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9293)
  TITLE     Mendenhall CRISPR tagging plasmids
REFERENCE   2  (bases 1 to 9293)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: PX458_ATF4_1
FEATURES             Location/Qualifiers
     source          1..9293
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        17..265
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     17..37
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     188..207
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        286..361
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        458..743
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        745..1022
                     /label=chicken beta-actin promoter
     intron          1023..1251
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1263..1272
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1272..1337
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1344..1364
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1389..5489
                     /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             1389..5489
                     /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             5490..5537
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5553..5606
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(5607..5609,5610..6320)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(5607..5609,5610..6320)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(5910..5929)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6257..6278
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6348..6365)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6354..6561
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6570..6710
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6570..6699
                     /label=AAV2 ITR
     rep_origin      6785..7240
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6872..6891)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7082..7103
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7257..7276)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7376..7398
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7436..7454)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7522..7626
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7627..7695,7696..8487)
                     /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(7845..7864)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8658..9246
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9147..9166
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttgctcac atgtnngagg gcctatttcc catgattcct tcatatttgc atatacgata
       61 caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca
      121 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
      181 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
      241 tatcttgtgg aaaggacgaa acaccgcccc tagttgagga tagtcgtttt agagctagaa
      301 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
      361 cttttttgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt ttttagcgcg
      421 tgcgccaatt ctgcagacaa atggctctag aggtacccgt tacataactt acggtaaatg
      481 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaatagta acgccaatag
      541 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      601 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      661 cctggcattg tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      721 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      781 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      841 cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
      901 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      961 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
     1021 cgggagtcgc tgcgacgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
     1081 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
     1141 cctccgggct gtaattagct gagcaagagg taagggttta agggatggtt ggttggtggg
     1201 gtattaatgt ttaattacct ggagcacctg cctgaaatca ctttttttca ggttggaccg
     1261 gtgccaccat ggactataag gaccacgacg gagactacaa ggatcatgat attgattaca
     1321 aagacgatga cgataagatg gccccaaaga agaagcggaa ggtcggtatc cacggagtcc
     1381 cagcagccga caagaagtac agcatcggcc tggacatcgg caccaactct gtgggctggg
     1441 ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg
     1501 accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag
     1561 ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga
     1621 tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc
     1681 acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct
     1741 tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga
     1801 gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg
     1861 cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca
     1921 gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa
     1981 accccatcaa cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga
     2041 gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg
     2101 gaaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg
     2161 ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc
     2221 tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg
     2281 ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg
     2341 cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg
     2401 tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct
     2461 acgccggcta cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca
     2521 tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc
     2581 tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc
     2641 tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa
     2701 agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa
     2761 acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg
     2821 aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg
     2881 ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca
     2941 ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct
     3001 tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag
     3061 tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg
     3121 aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga
     3181 aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata
     3241 tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct
     3301 atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct
     3361 ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa
     3421 tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg
     3481 acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata
     3541 gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc
     3601 agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca
     3661 tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg
     3721 agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac
     3781 accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg
     3841 ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg
     3901 accatatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca
     3961 gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga
     4021 tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca
     4081 atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga
     4141 gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga
     4201 tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga
     4261 agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca
     4321 acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca
     4381 aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc
     4441 ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct
     4501 acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga
     4561 agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg
     4621 attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg
     4681 aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc
     4741 tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg
     4801 tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga
     4861 gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca
     4921 tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc
     4981 ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg
     5041 aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg
     5101 ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg
     5161 tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga
     5221 gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg
     5281 ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg
     5341 gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca
     5401 ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac
     5461 ggatcgacct gtctcagctg ggaggcgaca aaaggccggc ggccacgaaa aaggccggcc
     5521 aggcaaaaaa gaaaaaggaa ttcggcagtg gagagggcag aggaagtctg ctaacatgcg
     5581 gtgacgtcga ggagaatcct ggcccagtga gcaagggcga ggagctgttc accggggtgg
     5641 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     5701 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     5761 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     5821 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     5881 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     5941 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     6001 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     6061 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     6121 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     6181 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     6241 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     6301 gcatggacga gctgtacaag gaattctaac tagagctcgc tgatcagcct cgactgtgcc
     6361 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     6421 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     6481 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     6541 gaatagcagg catgctgggg agcggccgca ggaaccccta gtgatggagt tggccactcc
     6601 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     6661 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat
     6721 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca
     6781 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     6841 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     6901 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     6961 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt
     7021 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     7081 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat
     7141 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     7201 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca
     7261 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     7321 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     7381 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     7441 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     7501 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     7561 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     7621 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     7681 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     7741 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     7801 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     7861 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     7921 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     7981 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     8041 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     8101 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     8161 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     8221 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     8281 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     8341 ggaagccgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     8401 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     8461 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     8521 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     8581 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     8641 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     8701 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     8761 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     8821 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     8881 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     8941 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     9001 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     9061 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     9121 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     9181 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     9241 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gcc
//
