LOCUS       sequence-375721-       10244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral delivery of Cas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10244)
  AUTHORS   Hosokawa H, Ungerback J, Wang X, Matsumoto M, Nakayama KI, Cohen SM,
            Tanaka T, Rothenberg EV
  TITLE     Transcription Factor PU.1 Represses and Activates Gene Expression in
            Early T Cells by Redirecting Partner Transcription Factor Binding.
  JOURNAL   Immunity. 2018 Jun 19;48(6):1119-1134.e7. doi: 
            10.1016/j.immuni.2018.04.024.
  PUBMED    29924977
REFERENCE   2  (bases 1 to 10244)
  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: 
            "10.1016/j.immuni.2018.04"; journalName: "Immunity"; date: 
            "2018-06-19- 19"; volume: "48"; issue: "6"; pages: "1119-1134.e7"
COMMENT     Sequence Label: pQCXIN-EF1a-mNeonGreen-P2A-Cas9
FEATURES             Location/Qualifiers
     source          1..10244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     124..143
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(224..241)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(395..983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(475..494)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1154..1945,1946..2014))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1777..1796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2015..2119)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2187..2205
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        2305..2684
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2685..2888
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2839..2859
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2889..3064
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    3127..3326
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             3527..3943
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3873..3895
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3911..3927
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     promoter        3978..4189
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      4213..4222
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4219..4926
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      4474..4483
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4942..4998
                     /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             5011..5031
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5032..9132
                     /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             5032..9132
                     /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             9133..9180
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     LTR             9326..9751
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(9854..9876)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(10012..10032)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(10017..10244,1..102)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(10197..10244,1..88)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
       61 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttac tggcttaact
      121 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
      181 gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc
      241 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
      301 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
      361 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      421 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      481 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      541 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
      601 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      661 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
      721 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
      781 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
      841 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
      901 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
      961 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1021 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1081 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1141 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1201 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     1261 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     1321 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1381 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1441 atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc tcgtcgtttg
     1501 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     1561 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     1621 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     1681 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     1741 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     1801 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     1861 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     1921 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     1981 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     2041 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     2101 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     2161 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag
     2221 aattagcttg gccattgcat acgttgtatc catatcataa tatgtacatt tatattggct
     2281 catgtccaac attaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa
     2341 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     2401 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2461 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2521 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2581 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2641 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2701 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2761 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2821 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2881 gcagagctca ataaaagagc ccacaacccc tcactcggcg cgccagtctt ccgatagact
     2941 gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg
     3001 tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccacgac gggggtcttt
     3061 catttggggg ctcgtccggg atttggagac ccctgcccag ggaccaccga cccaccaccg
     3121 ggaggtaagc tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgtttga
     3181 tgttatgcgc ctgcgtctgt actagttagc taactagctc tgtatctggc ggacccgtgg
     3241 tggaactgac gagttctgaa cacccggccg caaccctggg agacgtccca gggactttgg
     3301 gggccgtttt tgtggcccga cctgaggaag ggagtcgatg tggaatccga ccccgtcagg
     3361 atatgtggtt ctggtaggag acgagaacct aaaacagttc ccgcctccgt ctgaattttt
     3421 gctttcggtt tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct
     3481 gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aattagggcc agactgttac
     3541 cactccctta agtttgacct taggtcactg gaaagatgtc gagcggatcg ctcacaacca
     3601 gtcggtagat gtcaagaaga gacgttgggt taccttctgc tctgcagaat ggccaacctt
     3661 taacgtcgga tggccgcgag acggcacctt taaccgagac ctcatcaccc aggttaagat
     3721 caaggtcttt tcacctggcc cgcatggaca cccagaccag gtcccctaca tcgtgacctg
     3781 ggaagccttg gcttttgacc cccctccctg ggtcaagccc tttgtacacc ctaagcctcc
     3841 gcctcctctt cctccatccg ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc
     3901 tcgatcctcc ctttatccag ccctcactcc ttctctaggc gccggaattg aagatcttgg
     3961 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
     4021 aggggtcggc aattgaaccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
     4081 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
     4141 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg tgtcgtgacg
     4201 cggtctagaa tcgccaccat ggtgagcaag ggcgaggagg ataacatggc ctctctccca
     4261 gcgacacatg agttacacat ctttggctcc atcaacggtg tggactttga catggtgggt
     4321 cagggcaccg gcaatccaaa tgatggttat gaggagttaa acctgaagtc caccaagggt
     4381 gacctccagt tctccccctg gattctggtc cctcatatcg ggtatggctt ccatcagtac
     4441 ctgccctacc ctgacgggat gtcgcctttc caggccgcca tggtagatgg ctccggatac
     4501 caagtccatc gcacaatgca gtttgaagat ggtgcctccc ttactgttaa ctaccgctac
     4561 acctacgagg gaagccacat caaaggagag gcccaggtga aggggactgg tttccctgct
     4621 gacggtcctg tgatgaccaa ctcgctgacc gctgcggact ggtgcaggtc gaagaaaact
     4681 taccccaacg acaaaaccat catcagtacc tttaagtgga gttacaccac tggaaatggc
     4741 aagcgctacc ggagcactgc gcggaccacc tacacctttg ccaagccaat ggcggctaac
     4801 tatctgaaga accagccgat gtacgtgttc cgtaagacgg agctcaagca ctccaagacc
     4861 gagctcaact tcaaggagtg gcaaaaggcc tttaccgatg tgatgggcat ggacgagctg
     4921 tacaaggcgc gcggaagcgg agctactaac ttcagcctgc tgaagcaggc tggagacgtg
     4981 gaggagaacc ctggacctac gcgtatggcc ccaaagaaga agcggaaggt cgacaagaag
     5041 tacagcatcg gcctggacat cggcaccaac tctgtgggct gggccgtgat caccgacgag
     5101 tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag
     5161 aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc cacccggctg
     5221 aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag
     5281 atcttcagca acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc
     5341 ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac
     5401 gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac
     5461 agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc
     5521 cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg
     5581 ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc
     5641 ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat
     5701 ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct gattgccctg
     5761 agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg
     5821 cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac
     5881 cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac
     5941 atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga
     6001 tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct
     6061 gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacattgac
     6121 ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac
     6181 ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc
     6241 ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg
     6301 cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg
     6361 accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg
     6421 atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag
     6481 ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac
     6541 gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta taacgagctg
     6601 accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag
     6661 aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg
     6721 aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa
     6781 gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag
     6841 gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca
     6901 ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac
     6961 gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg
     7021 aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag
     7081 tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt
     7141 aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt
     7201 gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg
     7261 gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc
     7321 agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc
     7381 gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc
     7441 cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg
     7501 gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggaccatat cgtgcctcag
     7561 agctttctga aggacgactc catcgacaac aaggtgctga ccagaagcga caagaaccgg
     7621 ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg
     7681 cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac caaggccgag
     7741 agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc
     7801 cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac
     7861 gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc
     7921 gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc
     7981 cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg
     8041 gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag
     8101 agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac
     8161 tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc tctgatcgag
     8221 acaaacggcg aaaccgggga gatcgtgtgg gataagggcc gggattttgc caccgtgcgg
     8281 aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc
     8341 ttcagcaaag agtctatcct gcccaagagg aacagcgata agctgatcgc cagaaagaag
     8401 gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta ttctgtgctg
     8461 gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa agagctgctg
     8521 gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt tctggaagcc
     8581 aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta ctccctgttc
     8641 gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca gaagggaaac
     8701 gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca ctatgagaag
     8761 ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca gcacaagcac
     8821 tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat cctggccgac
     8881 gctaatctgg acaaagtgct gtccgcctac aacaagcacc gggataagcc catcagagag
     8941 caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc
     9001 aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac
     9061 gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag
     9121 ctgggaggcg acaagcgtcc tgctgctact aagaaagctg gtcaagctaa gaaaaagaaa
     9181 tgaatcgatt agtccaattt gttaaagaca ggatatcagt ggtccaggct ctagttttga
     9241 ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa taaaagattt
     9301 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     9361 cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata gagaagttca
     9421 gatcaaggtc aggaacagat ggaacagggt cgaccctaga gaaccatcag atgtttccag
     9481 ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt
     9541 ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca
     9601 ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc
     9661 ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg
     9721 attgactacc cgtcagcggg ggtctttcat ttgggggctc gtccgggatc gggagacccc
     9781 tgcccaggga ccaccgaccc accaccggga ggtaagctgg ctgcctcgcg cgtttcggtg
     9841 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     9901 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     9961 gcgcagccat gacccagtca cgtagcgata gcggagtgta gatccggctg tggaatgtgt
    10021 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
    10081 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
    10141 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
    10201 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggc
//
