LOCUS       sequence-207983-       13350 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  single-vector CROPseq system with blasticidin resistance selection 
            for use in the CROPseq CRISPR knockout screens.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13350)
  TITLE     CROPseq plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 13350)
  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            "Unpublished"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: CROPseq-EFS-SpCas9-P2A-blast
FEATURES             Location/Qualifiers
     source          1..13350
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1491..1535
                     /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    1716..1833
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2022..2233
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2252..2261
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2258..6361
                     /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             2258..6361
                     /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             6362..6409
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6410..6433
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6443..6499
                     /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             6500..6898
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    6914..7502
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6967..6987)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7385..7396)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        7658..7906
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7658..7678
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7829..7848
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9788..9863
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     LTR             9916..10096
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    10174..10295
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10211..10230)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10265..10284
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      10335..10470
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(10490..10509)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(10491..10509)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10519..10536)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(10519..10535)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10528..10550)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10677..11132
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10764..10783)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10974..10995
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        11158..11262
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(11263..11331,11332..12123)
                     /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(11481..11500)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      12294..12882
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12783..12802
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     13036..13053
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    13170..13191
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(13206..13211,13212..13229,13230..13236)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13244..13260
                     /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     13249..13271
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     13268..13284
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     13268..13284
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     13303..13323
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        13305..13323
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      301 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      361 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
      421 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
      481 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      541 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      601 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      661 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      721 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      781 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      841 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      901 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      961 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1021 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1081 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1141 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1201 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1261 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1321 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1381 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1441 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1501 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1561 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1621 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1681 agaatcttga gacaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
     1741 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     1801 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     1861 ccactttggc gccggctcga gggggcccgg gtgcaaagat ggataaagtt ttaaacagag
     1921 aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccgta
     1981 ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc
     2041 cacagtcccc gagaagttgg ggggaggggt cggcaattga tccggtgcct agagaaggtg
     2101 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
     2161 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
     2221 cgccagaaca caggaccggt tctagagcgc tgccaccatg gacaagaagt acagcatcgg
     2281 cctggacatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc
     2341 cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat
     2401 cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc
     2461 cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa
     2521 cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga
     2581 agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta
     2641 ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa
     2701 ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt
     2761 cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct
     2821 ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc
     2881 caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca
     2941 gctgcccggc gagaagaaga atggcctgtt cggaaacctg attgccctga gcctgggcct
     3001 gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa
     3061 ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga
     3121 cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt
     3181 gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca
     3241 ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa
     3301 agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag
     3361 ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga
     3421 actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg
     3481 cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga
     3541 tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat
     3601 cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa
     3661 gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc
     3721 ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct
     3781 gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa
     3841 atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat
     3901 cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta
     3961 cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa
     4021 cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga
     4081 caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga
     4141 cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat
     4201 gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa
     4261 cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt
     4321 cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat
     4381 ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc
     4441 cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt
     4501 gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca
     4561 gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat
     4621 caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa
     4681 cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact
     4741 ggacatcaac cggctgtccg actacgatgt ggaccatatc gtgcctcaga gctttctgaa
     4801 ggacgactcc atcgacaaca aggtgctgac cagaagcgac aagaaccggg gcaagagcga
     4861 caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc agctgctgaa
     4921 cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga gaggcggcct
     4981 gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac
     5041 aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg agaatgacaa
     5101 gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa
     5161 ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta
     5221 cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt
     5281 cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga
     5341 aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac
     5401 cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga
     5461 aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga aagtgctgag
     5521 catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga
     5581 gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg actgggaccc
     5641 taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa
     5701 agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat
     5761 catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa
     5821 agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa
     5881 cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct
     5941 gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc
     6001 ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact acctggacga
     6061 gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga
     6121 caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc aggccgagaa
     6181 tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca agtactttga
     6241 caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg ccaccctgat
     6301 ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggcga
     6361 caagcgacct gccgccacaa agaaggctgg acaggctaag aagaagaaag attacaaaga
     6421 cgatgacgat aagggatccg gcgcaacaaa cttctctctg ctgaaacaag ccggagatgt
     6481 cgaagagaat cctggaccga tggccaagcc tttgtctcaa gaagaatcca ccctcattga
     6541 aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc
     6601 agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg
     6661 accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac
     6721 ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg
     6781 acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca
     6841 gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaaac
     6901 gcgttaagtc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     6961 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     7021 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     7081 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     7141 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     7201 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     7261 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     7321 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     7381 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     7441 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     7501 gcgtcgactt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     7561 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagattt caccattatc
     7621 gtttcagacc cacctcccaa ccccgagggg acccagagag ggcctatttc ccatgattcc
     7681 ttcatatttg catatacgat acaaggctgt tagagagata attagaatta atttgactgt
     7741 aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt
     7801 tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta
     7861 tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccggag acggttgtaa
     7921 atgagcacac aaaatacaca tgctaaaata ttatattcta tgacctttat aaaatcaacc
     7981 aaaatcttct ttttaataac tttagtatca ataattagaa tttttatgtt cctttttgca
     8041 aacttttaat aaaaatgagc aaaataaaaa aacgctagtt ttagtaactc gcgttgtttt
     8101 cttcaccttt aataatagct actccaccac ttgttcctaa gcggtcagct cctgcttcaa
     8161 tcattttttg agcatcttca aatgttctaa ctccaccagc tgctttaact aaagcattgt
     8221 ctttaacaac tgacttcatt agtttaacat cttcaaatgt tgcacctgat tttgaaaatc
     8281 ctgttgatgt tttaacaaat tctaatccag cttcaacagc tatttcacaa gctttcatga
     8341 tttcttcttt tgttaataaa caattttcca taatacattt aacaacatgt gatccagctg
     8401 ctttttttac agctttcatg tcttctaaaa ctaattcata atttttgtct tttaatgcac
     8461 caatatttaa taccatatca atttctgttg caccatcttt aattgcttca gaaacttcga
     8521 atgcttttgt agctgttgtg catgcaccta gaggaaaacc tacaacattt gttattccta
     8581 catttgtgcc ttttaataat tctttacaat agcttgttca atatgaatta acacaaactg
     8641 ttgcaaaatc aaattcaatt gcttcatcac ataattgttt aatttcagct ttcgtagcat
     8701 cttgttttaa taatgtgtga tctatatatt tgtttagttt cattttttct cctatatatt
     8761 catttttaat tttaattctt taataatttc gtctacttta actttagcgt tttgaacaga
     8821 ttcaccaaca cctataaaat aaatttttag tttaggttca gttccacttg ggcgaacagc
     8881 aaatcatgac ttatcttcta aataaaattt tagtaagtct tgtcctggca tattatacat
     8941 tccatcgatg tagtcttcaa cattaacaac tttaagtcca gcaatttgag ttaagggtgt
     9001 tgctctcaat gatttcatta atggttcaat ttttaatttc ttttcttctg gtttaaaatt
     9061 caagtttaaa gtgaaagtgt aatatgcacc catttcttta aataaatctt ctaaatagtc
     9121 tactaatgtt ttattttgtt ttttataaaa tcaagcagcc tctgctatta atatagaagc
     9181 ttgtattcca tctttatctc tagctgagtc atcaattaca tatccataac tttcttcata
     9241 agcaaaaaca aaatttaatc cgttatcttc ttctttagca atttctctac ccattcattt
     9301 aaatccagtt aaagttttta caatattaac tccatatttt tcatgagcga ttctatcacc
     9361 caaatcactt gttacaaaac ttgaatatag agccggattt tttggaatgc tatttaagcg
     9421 ttttagattt gataattttc aatcaattaa aattggtcct gtttgatttc catctaatct
     9481 tacaaaatga ccatcatgtt ttattgccat tccaaatctg tcagcatctg ggtcattcat
     9541 aataataata tctgcatcat gtttaatacc atattcaagc ggtatttttc atgcaggatc
     9601 aaattctgga tttggattta caacattttt aaatgtttca tcttcaaatg catgctcttc
     9661 aacctcaata acgttatatc ctgattcacg taatattttt ggggtaaatt tagttcctgt
     9721 tccattaact gcgctaaaaa taatttttaa atctttttta gcttcttgct cttttttgta
     9781 cgtctctgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
     9841 aaaaagtggc accgagtcgg tgctttttta agcttggcgt aactagatct tgagacactg
     9901 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     9961 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
    10021 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
    10081 tggaaaatct ctagcagtac gtatagtagt tcatgtcatc ttattattca gtatttataa
    10141 cttgcaaaga aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt
    10201 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
    10261 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc
    10321 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
    10381 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
    10441 agaagtagtg aggaggcttt tttggaggcc tagggacgta cccaattcgc cctatagtga
    10501 gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
    10561 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
    10621 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc
    10681 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
    10741 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
    10801 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
    10861 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
    10921 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
    10981 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
    11041 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
    11101 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
    11161 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
    11221 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
    11281 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
    11341 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
    11401 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
    11461 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
    11521 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
    11581 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
    11641 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
    11701 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
    11761 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
    11821 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
    11881 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
    11941 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
    12001 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
    12061 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
    12121 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
    12181 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
    12241 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
    12301 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
    12361 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
    12421 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
    12481 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
    12541 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
    12601 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
    12661 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
    12721 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
    12781 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
    12841 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
    12901 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
    12961 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
    13021 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
    13081 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
    13141 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
    13201 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
    13261 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
    13321 agggaacaaa agctggagct gcaagcttaa
//
