LOCUS       sequence-435944-       11320 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of SpCas9-6xNLS(c-Myc) with puromycin 
            resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11320)
  AUTHORS   Shi J, et al.
  TITLE     Shi lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11320)
  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            "Unpublished"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: EFS-FLAG-SpCas9-6xNLS(c-Myc)-P2A-Puro (pRG214)
FEATURES             Location/Qualifiers
     source          1..11320
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    230..463
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             648..692
                     /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    876..993
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1062..1273
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      1293..1302
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1302..1325
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1326..5426
                     /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             1326..5426
                     /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             5436..5462
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             5472..5498
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             5508..5534
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             5544..5570
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             5580..5606
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             5616..5642
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             5658..5714
                     /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             5715..6314
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5715..5734)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6211..6231
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6330..6918
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6383..6403)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6801..6812)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6990..7223
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7301..7422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7338..7357)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7392..7411
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7462..7597
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7617..7636)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7618..7636)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7646..7663)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7646..7662)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7655..7677)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7804..8259
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7891..7910)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8101..8122
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8285..8389
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8390..8458,8459..9250)
                     /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(8608..8627)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9421..10009
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9910..9929
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10163..10180
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10297..10318
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10333..10338,10339..10356,10357..10363)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10371..10387
                     /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     10376..10398
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10395..10411
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10395..10411
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     10430..10450
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10432..10450
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        10478..10704
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             10705..10885
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    10932..11057
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
ORIGIN
        1 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
       61 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt
      121 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
      181 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
      241 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg
      301 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
      361 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
      421 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
      481 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
      541 tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca
      601 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag
      661 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc
      721 tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt
      781 ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga
      841 cccacctccc aaccccgagg ggacccagac cacaatttta aaagaaaagg ggggattggg
      901 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
      961 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     1021 ccactttggc gccggctcga gtggctccgg tgcccgtcag tgggcagagc gcacatcgcc
     1081 cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg
     1141 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
     1201 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     1261 cgccagaaca caggtgtcgt gacgcgggat ccgccaccat ggattacaaa gacgatgacg
     1321 ataaggacaa gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg
     1381 tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc
     1441 ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg
     1501 aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct
     1561 gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca
     1621 gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg
     1681 gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa
     1741 agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc
     1801 acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg
     1861 acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc
     1921 ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca
     1981 gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa
     2041 acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg
     2101 aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg
     2161 cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca
     2221 tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct
     2281 ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc
     2341 ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg
     2401 ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc
     2461 tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc
     2521 ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc
     2581 acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga
     2641 tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca
     2701 gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg
     2761 aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata
     2821 agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg
     2881 tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc
     2941 tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga
     3001 ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa
     3061 tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa
     3121 ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg
     3181 tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg
     3241 cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg
     3301 gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc
     3361 tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg
     3421 acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc
     3481 tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga
     3541 cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg
     3601 tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga
     3661 gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc
     3721 ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc
     3781 gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc
     3841 atatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa
     3901 gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga
     3961 agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc
     4021 tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac
     4081 agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga
     4141 acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt
     4201 ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca
     4261 actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa
     4321 agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga
     4381 agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca
     4441 gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc
     4501 ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt
     4561 ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg
     4621 tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga
     4681 tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc cccaccgtgg
     4741 cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg
     4801 tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg
     4861 actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta
     4921 agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac
     4981 tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca
     5041 gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg
     5101 aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag
     5161 tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata
     5221 agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag
     5281 cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca
     5341 aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga
     5401 tcgacctgtc tcagctggga ggcgacggcg gctcccctgc tgctaaacgt gttaagcttg
     5461 atgggggtag cccggcagcc aagagagtca aactcgacgg cggctcccct gcggcaaaaa
     5521 gggtgaaact agacgggggt agccccgccg cgaagcgtgt aaagctggat ggaggatcgc
     5581 ctgcggctaa gcgagtcaaa ttagacggtg ggtcaccggc tgcaaagcgc gtcaaattgg
     5641 atgctagcgg cagcggcgcc accaacttca gcctgctgaa gcaggccggc gacgtggagg
     5701 agaaccccgg ccccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     5761 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     5821 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     5881 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     5941 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     6001 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     6061 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg agtctcgccc gaccaccagg
     6121 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     6181 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     6241 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     6301 agcccggtgc ctgaacgcgt taagtcgaca atcaacctct ggattacaaa atttgtgaaa
     6361 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     6421 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     6481 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     6541 gcactgtgtt tgctgacgca acccccactg gttgggacat tgccaccacc tgtcagctcc
     6601 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     6661 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     6721 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     6781 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     6841 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     6901 tttgggccgc ctccccgcgt cgactttaag accaatgact tacaaggcag ctgtagatct
     6961 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     7021 agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
     7081 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     7141 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     7201 gtcagtgtgg aaaatctcta gcagtacgta tagtagttca tgtcatctta ttattcagta
     7261 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
     7321 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     7381 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
     7441 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     7501 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     7561 ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct
     7621 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     7681 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     7741 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     7801 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     7861 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     7921 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     7981 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     8041 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     8101 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     8161 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     8221 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     8281 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     8341 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     8401 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     8461 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     8521 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     8581 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     8641 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     8701 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     8761 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     8821 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     8881 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     8941 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     9001 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     9061 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     9121 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     9181 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     9241 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     9301 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     9361 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     9421 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     9481 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     9541 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     9601 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     9661 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     9721 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     9781 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     9841 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     9901 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     9961 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
    10021 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
    10081 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
    10141 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
    10201 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
    10261 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
    10321 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
    10381 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
    10441 tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg
    10501 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac
    10561 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag
    10621 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt
    10681 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag
    10741 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
    10801 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
    10861 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
    10921 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
    10981 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
    11041 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
    11101 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
    11161 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
    11221 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
    11281 atacagtagc aaccctctat tgtgtgcatc aaaggataga
//
