pJRH029

LOCUS       pJRH029               10504 bp ds-DNA     circular SYN 03-FEB-2023
DEFINITION  Plasmid for expressing Gag fused to spyCas9-3x FLAG-2x SV40 NLS with
            the SQNYPIVQ HIV-1 cleavage site in between.
ACCESSION   .
VERSION     .
KEYWORDS    pJRH029
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10504)
  AUTHORS   Hamilton JR, Tsuchida CA, Nguyen DN, Shy BR, McGarrigle ER, Sandoval
            Espinoza CR, Carr D, Blaeschke F, Marson A, Doudna JA
  TITLE     Targeted delivery of CRISPR-Cas9 and transgenes enables complex 
            immune cell engineering.
  JOURNAL   Cell Rep. 2021 Jun 1;35(9):109207. doi: 
            10.1016/j.celrep.2021.109207.
  PUBMED    34077734
REFERENCE   2  (bases 1 to 10504)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 3, 2023 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..10504
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1733..1742
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1739..3238
                     /codon_start=1
                     /gene="gag"
                     /product="gag protein from human immunodeficiency virus 1"
                     /label=HIV-1 gag
                     /translation="MGARASVLSGGELDRWEKIRLRPGGKKKYKLKHIVWASRELERFA
                     VNPGLLETSEGCRQILGQLQPSLQTGSEELRSLYNTVATLYCVHQRIEIKDTKEALDKI
                     EEEQNKSKKKAQQAAADTGHSNQVSQNYPIVQNIQGQMVHQAISPRTLNAWVKVVEEKA
                     FSPEVIPMFSALSEGATPQDLNTMLNTVGGHQAAMQMLKETINEEAAEWDRVHPVHAGP
                     IAPGQMREPRGSDIAGTTSTLQEQIGWMTHNPPIPVGEIYKRWIILGLNKIVRMYSPTS
                     ILDIRQGPKEPFRDYVDRFYKTLRAEQASQEVKNWMTETLLVQNANPDCKTILKALGPG
                     ATLEEMMTACQGVGGPGHKARVLAEAMSQVTNPATIMIQKGNFRNQRKTVKCFNCGKEG
                     HIAKNCRAPRKKGCWKCGKEGHQMKDCTERQANFLGKIWPSHKGRPGNFLQSRPEPTAP
                     PEESFRFGEETTTPSQKQEPIDKELYPLASLRSLFGSDPSSQ"
     CDS             3275..7378
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7382..7447
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7466..7486
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7493..7513
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(7526..7545)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7591..7646
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7645..7664)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(8007..8023)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8007..8023)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8020..8042)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8031..8047
                     /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)."
     promoter        complement(8055..8085)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8100..8121
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8180..8375
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8226..8361
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8288..8307
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    8381..8515
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8418..8437)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8472..8491
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8583..8600)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8754..9342)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8834..8853)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9513..10373)
                     /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     10136..10155
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10374..10478)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gccmggsggg gsggggsggg gsgrggggsg
      541 gggsggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tcgccaccat
     1741 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgatgggaaa aaattcggtt
     1801 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
     1861 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact
     1921 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
     1981 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
     2041 agacaagata gaggaagagc aaaacaaaag taagaaaaaa gcacagcaag cagcagctga
     2101 cacaggacac agcaatcagg tcagccaaaa ttaccctata gtgcagaaca tccaggggca
     2161 aatggtacat caggccatat cacctagaac tttaaatgca tgggtaaaag tagtagaaga
     2221 gaaggctttc agcccagaag tgatacccat gttttcagca ttatcagaag gagccacccc
     2281 acaagattta aacaccatgc taaacacagt ggggggacat caagcagcca tgcaaatgtt
     2341 aaaagagacc atcaatgagg aagctgcaga atgggataga gtgcatccag tgcatgcagg
     2401 gcctattgca ccaggccaga tgagagaacc aaggggaagt gacatagcag gaactactag
     2461 tacccttcag gaacaaatag gatggatgac acataatcca cctatcccag taggagaaat
     2521 ctataaaaga tggataatcc tgggattaaa taaaatagta agaatgtata gccctaccag
     2581 cattctggac ataagacaag gaccaaagga accctttaga gactatgtag accgattcta
     2641 taaaactcta agagccgagc aagcttcaca agaggtaaaa aattggatga cagaaacctt
     2701 gttggtccaa aatgcgaacc cagattgtaa gactatttta aaagcattgg gaccaggagc
     2761 gacactagaa gaaatgatga cagcatgtca gggagtgggg ggacccggcc ataaagcaag
     2821 agttttggct gaagcaatga gccaagtaac aaatccagct accataatga tacagaaagg
     2881 caattttagg aaccaaagaa agactgttaa gtgtttcaat tgtggcaaag aagggcacat
     2941 agccaaaaat tgcagggccc ctaggaaaaa gggctgttgg aaatgtggaa aggaaggaca
     3001 ccaaatgaaa gattgtactg agagacaggc taatttttta gggaagatct ggccttccca
     3061 caagggaagg ccagggaatt ttcttcagag cagaccagag ccaacagccc caccagaaga
     3121 gagcttcagg tttggggaag agacaacaac tccctctcag aagcaggagc cgatagacaa
     3181 ggaactgtat cctttagctt ccctcagatc actctttggc agcgacccct cgtcacaaaa
     3241 ctttagccag aactatccga ttgtgcagac cggtatggac aagaagtaca gcatcggcct
     3301 ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag
     3361 caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg
     3421 cgccctgctg ttcgacagcg gagaaacagc cgaggccacc cggctgaaga gaaccgccag
     3481 aagaagatac accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga
     3541 gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga
     3601 ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca
     3661 cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc
     3721 cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct
     3781 gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt
     3841 gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa
     3901 ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct
     3961 gcccggcgag aagaagaatg gcctgttcgg caacctgatt gccctgagcc tgggcctgac
     4021 ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga
     4081 cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct
     4141 gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa
     4201 caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca
     4261 ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga
     4321 gattttcttc gaccagagca agaacggcta cgccggctac atcgatggcg gagccagcca
     4381 ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact
     4441 gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag
     4501 catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt
     4561 ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc
     4621 ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag
     4681 cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg ccagcgccca
     4741 gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc
     4801 caagcacagc ctgctgtacg agtacttcac cgtgtacaac gagctgacca aagtgaaata
     4861 cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aagccatcgt
     4921 ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt
     4981 caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc
     5041 ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa
     5101 tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag
     5161 agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa
     5221 gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg
     5281 catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc
     5341 caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca
     5401 gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg
     5461 cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa
     5521 agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac
     5581 cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa
     5641 agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga
     5701 gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga
     5761 catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctgaagga
     5821 cgactccatc gataacaaag tgctgactcg gagcgacaag aaccggggca agagcgacaa
     5881 cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcgccagc tgctgaatgc
     5941 caagctgatt acccagagga agttcgacaa tctgaccaag gccgagagag gcggcctgag
     6001 cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa
     6061 gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga acgacaaact
     6121 gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga
     6181 tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct
     6241 gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt
     6301 gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat
     6361 cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga
     6421 gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac
     6481 aggcgagatc gtgtgggata agggccggga ctttgccacc gtgcggaaag tgctgtctat
     6541 gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc
     6601 tatcctgccc aagaggaaca gcgacaagct gatcgccaga aagaaggact gggaccctaa
     6661 gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt
     6721 ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat
     6781 ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga
     6841 agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg
     6901 ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc
     6961 ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc
     7021 cgaggataat gagcagaaac agctgtttgt ggaacagcac aaacactacc tggacgagat
     7081 catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa
     7141 ggtgctgagc gcctacaaca agcacagaga caagcctatc agagagcagg ccgagaatat
     7201 catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac
     7261 caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca
     7321 ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacgc
     7381 cgactataag gaccacgacg gagactacaa ggatcatgat attgattaca aagacgatga
     7441 cgataagagc ctgggcagcg gctcccccaa gaaaaaacgc aaggtggaag atcctaagaa
     7501 aaagcggaaa gtgtaagcta gcagatcttt ttccctctgc caaaaattat ggggacatca
     7561 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag
     7621 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa
     7681 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca
     7741 tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt
     7801 ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt
     7861 tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc
     7921 tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc
     7981 tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     8041 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     8101 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     8161 ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg cccctaactc
     8221 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     8281 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     8341 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta ttgcagctta
     8401 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     8461 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatccgctg
     8521 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     8581 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     8641 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     8701 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     8761 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     8821 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     8881 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     8941 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     9001 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     9061 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     9121 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     9181 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     9241 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     9301 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     9361 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     9421 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     9481 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     9541 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     9601 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     9661 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     9721 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     9781 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     9841 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     9901 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     9961 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
    10021 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
    10081 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
    10141 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
    10201 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
    10261 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
    10321 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
    10381 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
    10441 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
    10501 cctg
//