LOCUS       sequence-365327-       12810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiCRISPRv2 derivative with modified 7SK promoter driving sgRNA 
            and encoding Blasticidin resistance. Compatible with inserts 
            designed for lentiCRISPRv2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12810)
  AUTHORS   Hutcheson RL, Hayes M, Sugden B
  TITLE     Optimal LentiCRISPR-Based System for Sequential CRISPR/Cas9 Screens.
  JOURNAL   ACS Synth Biol. 2022 Jun 29. doi: 10.1021/acssynbio.2c00152.
  PUBMED    35767740
REFERENCE   2  (bases 1 to 12810)
  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: "ACS Synth 
            Biol. 2022 Jun 29. doi: 10.1021/acssynbio.2c00152."
COMMENT     Sequence Label: lentiCRISPRv2-7SK-Blast
FEATURES             Location/Qualifiers
     source          1..12810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_RNA        265..340
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        402..613
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      632..641
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             638..4741
                     /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             638..4741
                     /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             4742..4789
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4790..4813
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4823..4879
                     /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             4880..5275
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    5291..5879
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5344..5364)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5762..5773)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5951..6184
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(6210..6227)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6216..6440
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6486..6914
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6573..6592)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6783..6804
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6923..6943)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6928..7257
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7108..7243
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7170..7189
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7305..7352
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7371..7745
                     /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    7875..7996
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7912..7931)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7966..7985
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8045..8061)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8045..8061)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8058..8080)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8069..8085
                     /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(8093..8099,8100..8117,8118..8123))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8138..8159
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8276..8293)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8447..9035)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8527..8546)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9206..9997,9998..10066))
                     /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     9829..9848
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10067..10171)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10246..10265)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        10441..10820
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        10822..11020
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     10975..10995
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             11038..11218
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11265..11390
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11883..12116
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12301..12345
                     /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    12643..12760
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ctgcagtatt tagcatgccc cacccatctg caaggcattc tggatagtgt caaaacagcc
       61 ggaaatcaag tccgtttatc tcaaacttta gcattttggg aataaatgat atttgctatg
      121 ctggttaaat tagattttag ttaaatttcc tgctgaagct ctagtacgat aagcaacttg
      181 acctaagtgt aaagttgaga cttccttcag gtttatatag cttgtgcgcc gcttgggtac
      241 accggagacg gttaacacgt ctctgtttta gagctagaaa tagcaagtta aaataaggct
      301 agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttgaat tcgctagcta
      361 ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc
      421 cacagtcccc gagaagttgg ggggaggggt cggcaattga tccggtgcct agagaaggtg
      481 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
      541 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
      601 cgccagaaca caggaccggt tctagagcgc tgccaccatg gacaagaagt acagcatcgg
      661 cctggacatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc
      721 cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat
      781 cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc
      841 cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa
      901 cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga
      961 agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta
     1021 ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa
     1081 ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt
     1141 cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct
     1201 ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc
     1261 caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca
     1321 gctgcccggc gagaagaaga atggcctgtt cggaaacctg attgccctga gcctgggcct
     1381 gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa
     1441 ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga
     1501 cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt
     1561 gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca
     1621 ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa
     1681 agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag
     1741 ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga
     1801 actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg
     1861 cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga
     1921 tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat
     1981 cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa
     2041 gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc
     2101 ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct
     2161 gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa
     2221 atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat
     2281 cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta
     2341 cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa
     2401 cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga
     2461 caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga
     2521 cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat
     2581 gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa
     2641 cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt
     2701 cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat
     2761 ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc
     2821 cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt
     2881 gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca
     2941 gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat
     3001 caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa
     3061 cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact
     3121 ggacatcaac cggctgtccg actacgatgt ggaccatatc gtgcctcaga gctttctgaa
     3181 ggacgactcc atcgacaaca aggtgctgac cagaagcgac aagaaccggg gcaagagcga
     3241 caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc agctgctgaa
     3301 cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga gaggcggcct
     3361 gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac
     3421 aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg agaatgacaa
     3481 gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa
     3541 ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta
     3601 cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt
     3661 cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga
     3721 aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac
     3781 cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga
     3841 aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga aagtgctgag
     3901 catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga
     3961 gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg actgggaccc
     4021 taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa
     4081 agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat
     4141 catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa
     4201 agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa
     4261 cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct
     4321 gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc
     4381 ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact acctggacga
     4441 gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga
     4501 caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc aggccgagaa
     4561 tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca agtactttga
     4621 caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg ccaccctgat
     4681 ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggcga
     4741 caagcgacct gccgccacaa agaaggctgg acaggctaag aagaagaaag attacaaaga
     4801 cgatgacgat aagggatccg gcgcaacaaa cttctctctg ctgaaacaag ccggagatgt
     4861 cgaagagaat cctggaccgg ccaagccttt gtctcaagaa gaatccaccc tcattgaaag
     4921 agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc
     4981 tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc
     5041 ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg
     5101 tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca
     5161 ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc
     5221 gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg gctaaacgcg
     5281 ttaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     5341 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     5401 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     5461 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     5521 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     5581 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     5641 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     5701 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     5761 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     5821 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcg
     5881 tcgactttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     5941 aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt
     6001 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     6061 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     6121 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     6181 agcagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     6241 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     6301 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     6361 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     6421 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     6481 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     6541 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     6601 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     6661 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     6721 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     6781 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     6841 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     6901 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     6961 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     7021 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     7081 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     7141 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     7201 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     7261 ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat
     7321 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt
     7381 tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga
     7441 ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg
     7501 acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg
     7561 cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca
     7621 cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc
     7681 gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg
     7741 actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     7801 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     7861 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     7921 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     7981 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     8041 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     8101 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     8161 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     8221 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     8281 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     8341 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     8401 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     8461 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     8521 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     8581 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     8641 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     8701 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     8761 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     8821 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     8881 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     8941 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     9001 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     9061 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     9121 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     9181 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     9241 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     9301 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     9361 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     9421 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     9481 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     9541 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     9601 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     9661 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     9721 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     9781 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     9841 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     9901 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     9961 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
    10021 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
    10081 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
    10141 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
    10201 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
    10261 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
    10321 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
    10381 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgcgcgtt
    10441 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
    10501 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
    10561 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
    10621 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
    10681 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
    10741 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
    10801 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
    10861 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
    10921 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
    10981 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
    11041 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
    11101 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
    11161 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
    11221 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
    11281 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
    11341 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
    11401 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
    11461 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
    11521 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
    11581 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
    11641 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
    11701 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
    11761 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
    11821 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
    11881 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
    11941 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
    12001 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
    12061 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
    12121 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
    12181 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
    12241 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
    12301 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
    12361 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
    12421 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
    12481 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
    12541 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
    12601 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
    12661 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
    12721 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
    12781 cagagatcca gtttggttaa ttaaggtacc
//
