LOCUS       sequence-309981-       11326 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral Cas9 expression with CFP marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11326)
  TITLE     Lenitviral sgRNA and Cas9 constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11326)
  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: 
            "Unpublished"
COMMENT     Sequence Label: Cas9 CFP
FEATURES             Location/Qualifiers
     source          1..11326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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)."
     primer_bind     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     134..154
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        136..154
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        182..408
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             409..589
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    636..761
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1254..1487
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1672..1716
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1900..2017
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2086..2297
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2317..2326
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2326..2349
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2356..2376
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2401..6501
                     /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             2401..6501
                     /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             6502..6549
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6565..6621
                     /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             join(6622..6624,6625..6627,6628..7341)
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6928..6947)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7275..7296
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    7360..7948
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7413..7433)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7831..7842)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8020..8253
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8331..8452
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8368..8387)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8422..8441
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8492..8627
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8647..8666)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8648..8666)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8676..8693)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8676..8692)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8685..8707)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8834..9289
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8921..8940)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9131..9152
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9315..9419
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9420..9488,9489..10280)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(9638..9657)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10451..11039
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10940..10959
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11193..11210
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctta
      181 atgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag ttagcaacat
      241 gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag gtggtacgat
      301 cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac cactgaattg
      361 ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg gtctctctgg
      421 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
      481 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
      541 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
      601 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
      661 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
      721 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
      781 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
      841 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
      901 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
      961 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
     1021 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
     1081 agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac
     1141 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     1201 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     1261 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     1321 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     1381 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
     1441 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
     1501 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     1561 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     1621 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     1681 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     1741 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     1801 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     1861 cagacccacc tcccaacccc gaggggaccc agaccacaat tttaaaagaa aaggggggat
     1921 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     1981 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2041 agatccactt tggcgccggc tcgagtggct ccggtgcccg tcagtgggca gagcgcacat
     2101 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa
     2161 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     2221 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     2281 ttgccgccag aacacaggtg tcgtgacgcg ggatccgcca ccatggatta caaagacgat
     2341 gacgataaga tggccccaaa gaagaagcgg aaggtcggta tccacggagt cccagcagcc
     2401 gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc
     2461 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
     2521 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
     2581 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
     2641 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
     2701 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
     2761 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
     2821 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
     2881 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
     2941 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
     3001 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
     3061 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg
     3121 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
     3181 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
     3241 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
     3301 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
     3361 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
     3421 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
     3481 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
     3541 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
     3601 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
     3661 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
     3721 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
     3781 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
     3841 gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac
     3901 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat
     3961 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
     4021 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
     4081 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
     4141 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
     4201 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
     4261 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
     4321 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg
     4381 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
     4441 ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc
     4501 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
     4561 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
     4621 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
     4681 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg
     4741 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
     4801 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
     4861 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc
     4921 gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac
     4981 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
     5041 tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc
     5101 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
     5161 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
     5221 aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag
     5281 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
     5341 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
     5401 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
     5461 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac
     5521 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct
     5581 ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc
     5641 accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag
     5701 acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc
     5761 agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat
     5821 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa
     5881 gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt
     5941 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac
     6001 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag
     6061 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac
     6121 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag
     6181 cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc
     6241 ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc
     6301 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct
     6361 gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag
     6421 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac
     6481 ctgtctcagc tgggaggcga caagcgtcct gctgctacta agaaagctgg tcaagctaag
     6541 aaaaagaaag ctagcggcag cggcgccacc aacttcagcc tgctgaagca ggccggcgac
     6601 gtggaggaga accccggccc catggtgagc aagggcgagg agctgttcac cggggtggtg
     6661 cccatcctgg tcgagctgga cggcgacgta aacggccaca ggttcagcgt gtccggcgag
     6721 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     6781 ctgcccgtgc cctggcccac cctcgtgacc accctgacct ggggcgtgca gtgcttcagc
     6841 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     6901 gtccaggagc gtaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     6961 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     7021 gacggcaaca tcctggggca caagctggag tacaactaca tcagccacaa cgtctatatc
     7081 accgccgaca agcagaagaa cggcatcaag gcccacttca agatccgcca caacatcgag
     7141 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     7201 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     7261 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     7321 atggacgagc tgtacaagta atctacgcgt taagtcgaca atcaacctct ggattacaaa
     7381 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     7441 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     7501 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     7561 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     7621 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     7681 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     7741 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     7801 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     7861 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     7921 cggatctccc tttgggccgc ctccccgcgt cgactttaag accaatgact tacaaggcag
     7981 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     8041 aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     8101 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     8161 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     8221 gaccctttta gtcagtgtgg aaaatctcta gcagtacgta tagtagttca tgtcatctta
     8281 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta
     8341 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     8401 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     8461 ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     8521 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     8581 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc
     8641 aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt
     8701 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     8761 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     8821 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     8881 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     8941 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     9001 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     9061 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     9121 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     9181 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     9241 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     9301 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     9361 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     9421 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     9481 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     9541 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     9601 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     9661 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     9721 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     9781 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     9841 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     9901 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     9961 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
    10021 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
    10081 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
    10141 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
    10201 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
    10261 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
    10321 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
    10381 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
    10441 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
    10501 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
    10561 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
    10621 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
    10681 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
    10741 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
    10801 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
    10861 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
    10921 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
    10981 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
    11041 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
    11101 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
    11161 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
    11221 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
    11281 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaat
//
