LOCUS       sequence-171536-       13012 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentivirus carrying Cas9/CRISPR for cut in human PLXNB2 gene exon 3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13012)
  AUTHORS   Huang Y, Tejero R, Lee VK, Brusco C, Hannah T, Bertucci TB, 
            Junqueira Alves C, Katsyv I, Kluge M, Foty R, Zhang B, Friedel CC, 
            Dai G, Zou H, Friedel RH
  TITLE     Plexin-B2 facilitates glioblastoma infiltration by modulating cell 
            biomechanics.
  JOURNAL   Commun Biol. 2021 Jan 29;4(1):145. doi: 10.1038/s42003-021-01667-4.
  PUBMED    33514835
REFERENCE   2  (bases 1 to 13012)
  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: "Commun 
            Biol."; date: "2021-01-29"; volume: "4(1):145. doi"; pages: " 
            10.1038/s42003-021-01667-4"
COMMENT     Sequence Label: plentiCRISPRv2-sgPLXNB2
FEATURES             Location/Qualifiers
     source          1..13012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(16..36)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        21..350
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      201..336
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     263..282
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        398..445
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             464..838
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    968..1089
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1005..1024)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1059..1078
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1138..1154)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1138..1154)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1151..1173)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1162..1178
                     /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(1186..1192,1193..1210,1211..1216))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1231..1252
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1369..1386)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1540..2128)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1620..1639)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2299..3090,3091..3159))
                     /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     2922..2941
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3160..3264)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3339..3358)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3530..3909
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3911..4109
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4064..4084
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4127..4307
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4354..4479
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4972..5205
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5390..5434
                     /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    5732..5849
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5900..6148
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5900..5920
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6071..6090
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6169..6244
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        6306..6517
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      6536..6545
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6542..10645
                     /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             6542..10645
                     /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             10646..10693
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             10694..10717
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             10727..10783
                     /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             10784..11380
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     11277..11297
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    11396..11984
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11449..11469)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11867..11878)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             12056..12289
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(12315..12332)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12321..12545
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(12591..13012,1..7)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12678..12697)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12888..12909
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
       61 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
      121 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
      181 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
      241 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
      301 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
      361 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
      421 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag
      481 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
      541 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
      601 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
      661 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
      721 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
      781 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
      841 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
      901 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
      961 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     1021 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     1081 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     1141 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     1201 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     1261 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     1321 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     1381 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1441 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1501 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1561 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1621 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1681 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1741 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1801 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1861 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1921 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1981 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     2041 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     2101 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2161 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2221 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2281 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2341 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     2401 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     2461 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     2521 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     2581 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     2641 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     2701 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     2761 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     2821 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     2881 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     2941 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     3001 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     3061 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3121 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3181 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3241 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     3301 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
     3361 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     3421 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
     3481 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
     3541 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     3601 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     3661 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     3721 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     3781 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     3841 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     3901 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     3961 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     4021 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     4081 cgtgtacggt gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt
     4141 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     4201 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     4261 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac
     4321 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct
     4381 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact
     4441 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt
     4501 agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa
     4561 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa
     4621 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca
     4681 gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata
     4741 gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag
     4801 accaccgcac agcaagcggc cgctgatctt cagacctgga ggaggagata tgagggacaa
     4861 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc
     4921 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt
     4981 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac
     5041 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc
     5101 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc
     5161 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg
     5221 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc
     5281 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta
     5341 cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca
     5401 agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg
     5461 gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt
     5521 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca
     5581 gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg
     5641 agagagagac agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat
     5701 tctgcagaca aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt
     5761 acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac
     5821 aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag
     5881 tttggttaat taaggtaccg agggcctatt tcccatgatt ccttcatatt tgcatatacg
     5941 atacaaggct gttagagaga taattagaat taatttgact gtaaacacaa agatattagt
     6001 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
     6061 ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt
     6121 atatatcttg tggaaaggac gaaacaccgt tctcggcggc gaccgtcagt tttagagcta
     6181 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     6241 gtgctttttt gaattcgcta gctaggtctt gaaaggagtg ggaattggct ccggtgcccg
     6301 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
     6361 ttgatccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
     6421 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
     6481 cgttcttttt cgcaacgggt ttgccgccag aacacaggac cggttctaga gcgctgccac
     6541 catggacaag aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt
     6601 gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg
     6661 gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga
     6721 ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg
     6781 ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag
     6841 actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg
     6901 caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa
     6961 gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca
     7021 catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga
     7081 cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc
     7141 catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag
     7201 acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa
     7261 cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga
     7321 ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc
     7381 ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat
     7441 cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc
     7501 tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg
     7561 gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc
     7621 cggctacatt gacggcggag ccagccagga agagttctac aagttcatca agcccatcct
     7681 ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg
     7741 gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca
     7801 cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat
     7861 cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag
     7921 cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga
     7981 agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa
     8041 gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt
     8101 gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct
     8161 gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac
     8221 cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat
     8281 ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat
     8341 tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt
     8401 gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc
     8461 ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg
     8521 caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct
     8581 ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga
     8641 cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct
     8701 gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac
     8761 agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt
     8821 gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag
     8881 aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc
     8941 cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg
     9001 ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca
     9061 tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag
     9121 cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa
     9181 gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct
     9241 gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca
     9301 gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa
     9361 cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc
     9421 caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa
     9481 ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa
     9541 gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa
     9601 gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag
     9661 caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg
     9721 gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt
     9781 tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt
     9841 gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat
     9901 cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc
     9961 ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt
    10021 gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga
    10081 ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa
    10141 gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact
    10201 gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag
    10261 ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga
    10321 acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt
    10381 gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa
    10441 gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc
    10501 ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa
    10561 agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat
    10621 cgacctgtct cagctgggag gcgacaagcg acctgccgcc acaaagaagg ctggacaggc
    10681 taagaagaag aaagattaca aagacgatga cgataaggga tccggcgcaa caaacttctc
    10741 tctgctgaaa caagccggag atgtcgaaga gaatcctgga ccgaccgagt acaagcccac
    10801 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
    10861 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
    10921 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
    10981 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
    11041 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
    11101 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
    11161 cgtcggagtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
    11221 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
    11281 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
    11341 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga acgcgttaag tcgacaatca
    11401 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
    11461 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
    11521 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
    11581 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
    11641 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
    11701 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
    11761 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
    11821 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
    11881 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
    11941 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcgtcgac tttaagacca
    12001 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
    12061 gggctaattc actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg
    12121 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
    12181 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
    12241 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag ggcccgttta
    12301 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
    12361 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
    12421 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
    12481 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
    12541 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
    12601 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
    12661 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
    12721 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
    12781 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
    12841 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
    12901 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
    12961 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta ac
//
