LOCUS       sequence-451031-       10783 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MMLVgag(C507V)–Cas9 for producing v5 Cas9-eVLPs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10783)
  AUTHORS   Raguram A, An M, Chen PZ, Liu DR
  TITLE     Directed evolution of engineered virus-like particles with improved 
            production and transduction efficiencies.
  JOURNAL   Nat Biotechnol. 2024 Nov 13. doi: 10.1038/s41587-024-02467-x.
  PUBMED    39537813
REFERENCE   2  (bases 1 to 10783)
  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"; journalName: "Nat 
            Biotechnol. 2024 Nov 13. doi: 10.1038/s41587-024-02467-x."
COMMENT     Sequence Label: v5 Cas9-eVLP
FEATURES             Location/Qualifiers
     source          1..10783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4..420
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     350..372
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     388..404
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1633..1659
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             1672..1698
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             1711..1737
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             1771..1794
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1828..1848
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1861..5961
                     /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             1861..5961
                     /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             6004..6024
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6030..6049)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    6095..6150
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(6149..6168)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6508..6524)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6508..6524)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6521..6543)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6532..6548
                     /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(join(6556..6562,6563..6580,6581..6586))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6601..6622
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6739..6756)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6910..7498)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6990..7009)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7669..8460,8461..8529))
                     /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     8292..8311
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8530..8634)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8660..9115)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8797..8818)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9009..9028
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9242..9264
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9256..9273
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9257..9273
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9283..9302
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9283..9301
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        9422..9801
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9802..10005
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9956..9976
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     10002..10026
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          10117..10689
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
ORIGIN
        1 atgggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa agatgtcgag
       61 cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac cttctgctct
      121 gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc
      181 atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc agaccaggtc
      241 ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt caagcccttt
      301 gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc ccttgaacct
      361 cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc tctaggcgcc
      421 aaacctaaac ctcaagttct ttctgacagt ggggggccgc tcatcgacct acttacagaa
      481 gaccccccgc cttataggga cccaagacca cccccttccg acagggacgg aaatggtgga
      541 gaagcgaccc ctgcgggaga ggcaccggac ccctccccaa tggcatctcg cctacgtggg
      601 agacgggagc cccctgtggc cgactccact acctcgcagg cattccccct ccgcgcagga
      661 ggaaacggac agcttcaata ctggccgttc tcctcttctg acctttacaa ctggaaaaat
      721 aataaccctt ctttttctga agatccaggt aaactgacag ctctgatcga gtctgtcctc
      781 atcacccatc agcccacctg ggacgactgt cagcagctgt tggggactct gctgaccgga
      841 gaagaaaaac aacgggtgct cttagaggct agaaaggcgg tgcggggcga tgatgggcgc
      901 cccactcaac tgcccaatga agtcgatgcc gcttttcccc tcgagcgccc agactgggat
      961 tacaccaccc aggcaggtag gaaccaccta gtccactatc gccagttgct cctagcgggt
     1021 ctccaaaacg cgggcagaag ccccaccaat ttggccaagg taaaaggaat aacacaaggg
     1081 cccaatgagt ctccctcggc cttcctagag agacttaagg aagcctatcg caggtacact
     1141 ccttatgacc ctgaggaccc agggcaagaa actaatgtgt ctatgtcttt catttggcag
     1201 tctgccccag acattgggag aaagttagag aggttagaag atttaaaaaa caagacgctt
     1261 ggagatttgg ttagagaggc agaaaagatc tttaataaac gagaaacccc ggaagaaaga
     1321 gaggaacgta tcaggagaga aacagaggaa aaagaagaac gccgtaggac agaggatgag
     1381 cagaaagaga aagaaagaga tcgtaggaga catagagaga tgagcaagct attggccact
     1441 gtcgttagtg gacagaaaca ggatagacag ggaggagaac gaaggaggtc ccaactcgat
     1501 cgcgaccagt gtgcctacgt gaaagaaaag gggcactggg ctaaagattg tcccaagaaa
     1561 ccacgaggac ctcggggacc aagaccccag acctccctcc tgaccctaga tgactctggc
     1621 ggctcacttc aactgcctcc acttgaaaga ctgacactgg gatcattaca attacctcct
     1681 ttagaacgat taacactcgg ttcactacag cttccgcctc ttgagagatt gacattaacg
     1741 tccacgctgc taatggagaa ctcgtctgga gattacaaag acgatgacga taagaaacgg
     1801 acagccgacg gaagcgagtt cgagtcacca aagaagaagc ggaaagtcag cggggggtca
     1861 gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc
     1921 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
     1981 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
     2041 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
     2101 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
     2161 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
     2221 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
     2281 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
     2341 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
     2401 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
     2461 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
     2521 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg
     2581 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
     2641 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
     2701 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
     2761 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
     2821 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
     2881 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
     2941 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
     3001 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
     3061 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
     3121 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
     3181 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
     3241 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
     3301 gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac
     3361 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat
     3421 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
     3481 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
     3541 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
     3601 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
     3661 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
     3721 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
     3781 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg
     3841 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
     3901 ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc
     3961 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
     4021 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
     4081 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
     4141 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg
     4201 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
     4261 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
     4321 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc
     4381 gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac
     4441 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
     4501 tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc
     4561 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
     4621 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
     4681 aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag
     4741 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
     4801 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
     4861 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
     4921 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac
     4981 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct
     5041 ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc
     5101 accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag
     5161 acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc
     5221 agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat
     5281 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa
     5341 gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt
     5401 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac
     5461 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag
     5521 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac
     5581 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag
     5641 cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc
     5701 ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc
     5761 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct
     5821 gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag
     5881 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac
     5941 ctgtctcagc tgggaggtga ctctggcggc tcaaaaagaa ccgccgacgg cagcgaattc
     6001 gagcccaaga agaagaggaa agtctgagat ctttttccct ctgccaaaaa ttatggggac
     6061 atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca
     6121 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat
     6181 ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct
     6241 gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc
     6301 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt
     6361 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt
     6421 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc
     6481 gacggatcgg ccgcaattcg taaccatgtc atagctgttt cctgtgtgaa attgttatcc
     6541 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6601 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6661 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6721 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6781 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6841 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6901 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6961 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7021 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7081 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     7141 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7201 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7261 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7321 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     7381 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7441 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7501 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7561 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7621 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7681 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7741 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7801 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7861 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7921 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7981 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8041 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8101 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8161 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8221 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8281 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8341 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8401 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8461 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8521 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8581 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8641 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     8701 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     8761 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     8821 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     8881 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     8941 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     9001 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     9061 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     9121 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     9181 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     9241 tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag
     9301 ggcgaattgg agctccaccg cggtggcggc cgctctagag agcttggccc attgcatacg
     9361 ttgtatccat atcataatat gtacatttat attggctcat gtccaacatt accgccatgt
     9421 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     9481 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     9541 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     9601 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     9661 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     9721 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     9781 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     9841 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     9901 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     9961 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt
    10021 cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga
    10081 tccagcctcc ggtcgaccga tcctgagaac ttcagggtga gtttggggac ccttgattgt
    10141 tctttctttt tcgctattgt aaaattcatg ttatatggag ggggcaaagt tttcagggtg
    10201 ttgtttagaa tgggaagatg tcccttgtat caccatggac cctcatgata attttgtttc
    10261 tttcactttc tactctgttg acaaccattg tctcctctta ttttcttttc attttctgta
    10321 actttttcgt taaactttag cttgcatttg taacgaattt ttaaattcac ttttgtttat
    10381 ttgtcagatt gtaagtactt tctctaatca cttttttttc aaggcaatca gggtatatta
    10441 tattgtactt cagcacagtt ttagagaaca attgttataa ttaaatgata aggtagaata
    10501 tttctgcata taaattctgg ctggcgtgga aatattctta ttggtagaaa caactacacc
    10561 ctggtcatca tcctgccttt ctctttatgg ttacaatgat atacactgtt tgagatgagg
    10621 ataaaatact ctgagtccaa accgggcccc tctgctaacc atgttcatgc cttcttctct
    10681 ttcctacagc tcctgggcaa cgtgctggtt gttgtgctgt ctcatcattt tggcaaagaa
    10741 ttcctcgagg gccggatccg aattcctgta tttgtctgaa aac
//
