LOCUS       sequence-413204-       11302 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9-RC and GFP from an EF1-alpha promoter. High-performance knockin
            through DSB repair by homology-directed recombination (HDR) or 
            homology-mediated end joining (HMEJ). [Lab plasmid ID: N213].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11302)
  AUTHORS   Richardson RR, Steyert M, Khim SN, Crutcher GW, Brandenburg C, 
            Robertson CD, Romanowski AJ, Inen J, Altas B, Poulopoulos A
  TITLE     Enhancing Precision and Efficiency of Cas9-Mediated Knockin Through 
            Combinatorial Fusions of DNA Repair Proteins.
  JOURNAL   CRISPR J. 2023 Oct;6(5):447-461. doi: 10.1089/crispr.2023.0036. Epub
            2023 Sep 15.
  PUBMED    37713292
REFERENCE   2  (bases 1 to 11302)
  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"; doi: 
            "10.1089/crispr.2023.0036"; journalName: "CRISPR J"; date: 
            "2023-10"; volume: "6"; issue: "5"; pages: "447-461"
COMMENT     Sequence Label: pS34.EF1-alpha<GFP-2A-Cas9-RC
FEATURES             Location/Qualifiers
     source          1..11302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(247..835)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(327..346)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1006..1797,1798..1866))
                     /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     1629..1648
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1867..1971)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2661..2872
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          2873..3811
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3768..3788
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3857..3866
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3863..3865,3866..3868,3869..4579)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3863..3865,3866..3868,3869..4579)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4169..4188)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4516..4537
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             4595..4651
                     /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             6029..6049
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6074..10174
                     /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             6074..10174
                     /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             10175..10222
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(11204..11223)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11258..11277
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ggtgaattcc cttatgtgag tgtaaaaggc aggcgagttt gtcccttaat taatagggat
       61 aacagggtaa tatcttgccc tgcacgaata ccagcgaccc cttgcccaaa tacttgccgt
      121 gggcctcggc ctgagagcca aaacacttga tgcggaagaa gtcggtgcgc tcctgcttgt
      181 cgccggcatc gttgcgccac atctaggatc tgccaggaac cgtaaaaagg ccgcgttgct
      241 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      301 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      361 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      421 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      481 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      541 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      601 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      661 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
      721 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
      781 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
      841 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
      901 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
      961 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1021 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1081 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1141 accgcgagaa ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1201 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1261 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1321 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1381 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     1441 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1501 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1561 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     1621 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     1681 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     1741 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     1801 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     1861 actcatactc ttcctttttc aatattattg aagcatttat caaggttatt gtctcatgag
     1921 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacgaattg
     1981 gccagcgctg cattaccctg ttatccctaa ccggtctaat acccaatctc tcgtcttatc
     2041 cagatgtttt atacgccgga gttctgcaat tcatgtgatc tgcgcctggg agctcacggg
     2101 gacagccccc ccccaaagcc cccagggatg taattacgtc cctcccccgc tagggggcag
     2161 cagcgagccg cccggggctc cgctccggtc cggcgctccc cccgcatccc cgagccggca
     2221 gcgtgcgggg acagcccggg cacggggaag gtggcacggg atcgctttcc tctgaacgct
     2281 tctcgctgct ctttgagcct gcagacacct ggggggatac ggggaaaaag ctaggctgaa
     2341 agaggatctt agaatgacac gcgcctggga gctcacgggg acagcccccc cccaaagccc
     2401 ccagggatgt aattacgtcc ctcccccgct agggggcagc agcgagccgc ccggggctcc
     2461 gctccggtcc ggcgctcccc ccgcatcccc gagccggcag cgtgcgggga cagcccgggc
     2521 acggggaagg tggcacggga tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg
     2581 cagacacctg gggggatacg gggaaaaagc tggatcataa ctcgacgtgc atactttaat
     2641 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttgtg
     2701 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
     2761 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
     2821 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
     2881 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
     2941 tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg
     3001 ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg
     3061 gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc
     3121 tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg
     3181 gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc
     3241 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg
     3301 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc
     3361 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc
     3421 agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag
     3481 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc
     3541 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga
     3601 ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat
     3661 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta
     3721 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac
     3781 agtggttcaa agtttttttc ttccatttca ggtggatccc aagggaatgt aagtttgtac
     3841 aaaaaagaga ggtgccgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     3901 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     3961 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     4021 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     4081 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     4141 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     4201 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     4261 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     4321 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     4381 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     4441 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     4501 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     4561 catggacgag ctgtacaagg gagccggaag cggagctact aacttcagcc tgctgaagca
     4621 ggctggcgat gtggaagaga accctggacc tgctggagac tccctggccg agtctcggtg
     4681 gcctccgggc ctggcagtca tgaagacaat agatgatttg ctgcggtgtg gaatttgctt
     4741 cgagtatttc aacattgcaa tgataatacc tcagtgttca cataactact gctctctctg
     4801 tataagaaaa tttctgtcct ataaaactca gtgtccaact tgctgtgtga ctgtcacaga
     4861 gccggatctg aaaaataacc gcatattaga tgaactggta aaaagcttga attttgcacg
     4921 gaatcatctg ctgcagtttg ctttagagtc accagccaaa tctcctgctt cttcctcttc
     4981 aaagaatctt gctgtcaaag tatatactcc tgtagcctcc agacagtctt taaagcaggg
     5041 gagcaggtta atggataatt tcttgatcag agaaatgagt ggttctacat cagagttgtt
     5101 gataaaagaa aataaaagca aattcagccc tcaaaaagag gcgagccctg ctgcaaagac
     5161 caaagagaca cgttctgtag aagagatcgc tccagatccc tcagaggcta agcgtcctga
     5221 gccaccctcg acatccactt tgaaacaagt tactaaagtg gattgtcctg tttgcggggt
     5281 taacattcca gaaagtcaca ttaataagca tttagacagc tgtttatcac gcgaagagaa
     5341 gaaggaaagc ctcagaagtt ctgttcacaa aaggaagccg cacatgtaca atgcccaatg
     5401 cgatgctttg catcctaaat cagctgctga aatagttcaa gaaatcgaaa atatagagaa
     5461 gactaggatg cgtcttgaag ctagtaaact caatgaaagt gtaatggttt ttacaaagga
     5521 ccaaacagaa aaggaaatag atgaaatcca cagtaaatat cgtaaaaaac ataagagtga
     5581 atttcagctt ctggtggatc aggctagaaa aggatacaag aaaattgctg gaatgtcaca
     5641 aaaaacagta acaataacaa aagaagatga atctacagaa aagctatctt ctgtatgcat
     5701 gggacaggaa gataatatga cctcagtaac aaaccacttt tctcaatcaa agctggactc
     5761 cccagaggaa ttggaacctg acagagaaga ggattcttct agctgtattg atattcaaga
     5821 agttctttct tcatcagaat cagattcatg caatagttcc agttcagaca tcataagaga
     5881 tcttttagaa gaagaggaag cctgggaagc atcacataaa aacgatcttc aagacacaga
     5941 aataagtcca agacagaatc gccgcacaag agccgctgaa agtgctgaga ttgaaccaag
     6001 aaacaagcgt aataggaatg gagcagcccc aaagaagaag cggaaggtcg gtatccacgg
     6061 agtcccagca gccgacaaga agtacagcat cggcctggac atcggcacca actctgtggg
     6121 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
     6181 caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga
     6241 aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa
     6301 ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt
     6361 cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc
     6421 catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca
     6481 cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc
     6541 cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga
     6601 caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga
     6661 ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag
     6721 caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct
     6781 gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga
     6841 cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa
     6901 cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc
     6961 cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct
     7021 gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc
     7081 tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa
     7141 cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa
     7201 gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga
     7261 cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg
     7321 agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg
     7381 ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag
     7441 gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa
     7501 cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa
     7561 cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta
     7621 cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc
     7681 cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg
     7741 gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc
     7801 cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct
     7861 gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga
     7921 agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa
     7981 aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac
     8041 cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa
     8101 gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat
     8161 ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg
     8221 cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat
     8281 cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga
     8341 gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag
     8401 ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa
     8461 agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca
     8521 gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga
     8581 tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct
     8641 gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa
     8701 gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt
     8761 cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat
     8821 caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc
     8881 ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac
     8941 cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga
     9001 gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct
     9061 gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga
     9121 cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt
     9181 cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat
     9241 ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg
     9301 ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa
     9361 gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga
     9421 taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc
     9481 caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact
     9541 gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa
     9601 tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa
     9661 gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc
     9721 cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta
     9781 cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct
     9841 gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc
     9901 caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca
     9961 ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa
    10021 tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac
    10081 cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga
    10141 gacacggatc gacctgtctc agctgggagg cgacaaaagg ccggcggcca cgaaaaaggc
    10201 cggccaggca aaaaagaaaa aggaattcgg cagtggagga gccgctagca tgaacatctc
    10261 gggaagcagc tgtggaagcc ctaactctgc agatacatct agtgacttta aagatctgtg
    10321 gaccaaacta aaagaatgtc atgatagaga agtacaaggt ttacaagtaa aagtaaccaa
    10381 gcttaagcaa gagcgaatct tagatgcaca aagactagaa gaattcttca ccaaaaatca
    10441 acagctgagg gaacagcaga aagtccttca tgaagccatt aaagttttag aagatcggtt
    10501 aagagcaggc ttatgtgatc gctgtgcagt aactgaagaa catatgcgga aaaaacagca
    10561 agagtttgaa aatatccggc agcagaatct taaacttatt acagaactta tgaatgaaag
    10621 gaatactcta caggaagaaa ataaaaagct ttctgaacaa ctccagcaga aaattgagaa
    10681 tgatcaacag catcaagcag ctgagcttga atgtgaggaa gacgttattc cagattcacc
    10741 gataacagcc ttctcatttt ctggcgttaa ccggctacga agaaaggaga acccccatgt
    10801 ccgatacata gaacaaacac atactaaatt ggagcactct gtgtgtgcaa atgaaatgag
    10861 aaaagtttcc aagtcttcaa ctcatccaca acataatcct aatgaaaatg aaattctagt
    10921 agctgacact tatgaccaaa gtcaatctcc aatggccaaa gcacatggaa caagcagcta
    10981 tacccctgat aagtcatctt ttaatttagc tacagttgtt gctgaaacac ttggacttgg
    11041 tgttcaagaa gaatctgaaa ctcaaggtcc catgagcccc cttggtgatg agctctacca
    11101 ctgtctggaa ggaaatcaca agaaacagcc ttttgagtag cttccactgg attgtacaat
    11161 taccaacact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
    11221 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
    11281 gtatcttatc atgtctgtcg ct
//
