LOCUS       sequence-388043-       14583 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  WT SpCas9 with BSD-TagBFP for selection and sorting.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14583)
  TITLE     Schmierer unpublished plasmids
REFERENCE   2  (bases 1 to 14583)
  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"
COMMENT     Sequence Label: lenti.Cas9.BFP.Blast
FEATURES             Location/Qualifiers
     source          1..14583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(19..607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(99..118)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(778..1569,1570..1638))
                     /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     1401..1420
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1639..1743)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1818..1837)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2009..2388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2390..2588
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2543..2563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2606..2786
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2833..2958
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3451..3684
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3869..3913
                     /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    4211..4328
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        4458..5636
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4689..5627
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5584..5604
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             5652..5699
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5700..9800
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5700..9800
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             9807..9827
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9852..9905
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             9906..10301
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             10323..11021
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     misc_feature    11046..11634
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11099..11119)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(11517..11528)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(11637..11653)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11638..11654)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             12159..12339
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(12365..12382)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12371..12595
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      12641..13069
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12728..12747)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12938..12959
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(13078..13098)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        13083..13412
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      13263..13398
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     13325..13344
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        13460..13507
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             13526..13900
                     /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    14030..14151
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(14067..14086)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     14121..14140
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(14200..14216)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(14200..14216)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14213..14235)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    14224..14240
                     /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(14248..14254,14255..14272,14273..14278))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    14293..14314
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(14431..14448)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
       61 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      121 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      181 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      241 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      301 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      361 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
      421 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
      481 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
      541 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
      601 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
      661 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
      721 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
      781 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
      841 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
      901 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
      961 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1021 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1081 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1141 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1201 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1261 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1321 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1381 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1441 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1501 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     1561 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     1621 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     1681 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     1741 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     1801 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     1861 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     1921 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     1981 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
     2041 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     2101 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     2161 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     2221 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     2281 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     2341 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     2401 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     2461 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     2521 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     2581 ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga
     2641 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     2701 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     2761 ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg
     2821 aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc
     2881 gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga
     2941 gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt
     3001 cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg
     3061 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa
     3121 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat
     3181 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa
     3241 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc
     3301 gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg aattatataa
     3361 atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt
     3421 ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc
     3481 agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt
     3541 gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct
     3601 gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag
     3661 atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac
     3721 cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca
     3781 cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt
     3841 aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa
     3901 atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt
     3961 cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt
     4021 gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag
     4081 gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc
     4141 cattcgatta gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa atggcagtat
     4201 tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
     4261 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
     4321 aaaattttcg ggtttattac agggacagca gagatccagt ttggttaatt agctagctgc
     4381 aaagatggat aaagttttaa acagagagga atctttgcag ctaatggacc ttctaggtct
     4441 tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
     4501 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
     4561 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
     4621 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
     4681 gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc
     4741 cttgcgtgcc ttgaattact tccacctggc tgcagtacgt gattcttgat cccgagcttc
     4801 gggttggaag tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg
     4861 cttgagttga ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc
     4921 gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg
     4981 cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta
     5041 tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg
     5101 cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc
     5161 ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc
     5221 tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag
     5281 ggagctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa
     5341 ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc
     5401 cgtccaggca cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg
     5461 aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag
     5521 cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca
     5581 ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgacgta
     5641 cggccaccat gaaaaggccg gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagg
     5701 acaagaagta cagcatcggc ctggacatcg gcaccaactc tgtgggctgg gccgtgatca
     5761 ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca
     5821 gcatcaagaa gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca
     5881 cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc
     5941 tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg
     6001 aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca
     6061 tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac
     6121 tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga
     6181 tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg
     6241 acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca
     6301 acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc
     6361 tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggaaacctga
     6421 ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg
     6481 ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga
     6541 tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc
     6601 tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga
     6661 tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc
     6721 agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct
     6781 acattgacgg cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa
     6841 agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc
     6901 agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca
     6961 ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga
     7021 agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat
     7081 tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg
     7141 tggacaaggg cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc
     7201 tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata
     7261 acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg
     7321 gcgagcagaa aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga
     7381 agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg
     7441 gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca
     7501 aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga
     7561 ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc
     7621 tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc
     7681 tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt
     7741 tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc
     7801 tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg
     7861 agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga
     7921 aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg
     7981 aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga
     8041 agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg
     8101 aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata
     8161 tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gaccatatcg
     8221 tgcctcagag ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca
     8281 agaaccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact
     8341 actggcggca gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca
     8401 aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg
     8461 tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta
     8521 agtacgacga gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc
     8581 tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc
     8641 accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc
     8701 ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga
     8761 tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca
     8821 tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc
     8881 tgatcgagac aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca
     8941 ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga
     9001 caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgataag ctgatcgcca
     9061 gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt
     9121 ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag
     9181 agctactggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc
     9241 tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact
     9301 ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga
     9361 agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact
     9421 atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc
     9481 acaagcacta cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc
     9541 tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca
     9601 tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg
     9661 ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg
     9721 tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc
     9781 tgtctcagct gggaggcgac agcggaccta agaaaaagag gaaggtggcg gccgctggat
     9841 ccggcagtgg agagggcaga ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg
     9901 gcccaatggc caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta
     9961 caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg
    10021 acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac
    10081 tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga
    10141 tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg
    10201 atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg
    10261 ggattcgtga attgctgccc tctggttatg tgtgggaggg cgctagcggc agtggaagtg
    10321 gcagcgagct gattaaggag aacatgcaca tgaagctcta catggagggc accgtggaca
    10381 accatcactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca
    10441 tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacta
    10501 gcttcctcta cggcagcaag accttcatca accacaccca gggcatcccc gacttcttca
    10561 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg
    10621 tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga
    10681 tcagaggggt gaacttcaca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg
    10741 aggccttcac cgaaacgctg taccccgctg acggcggcct ggaaggcaga aacgacatgg
    10801 ccctgaagct cgtgggcggg agccatctga tcgcaaacat caagaccaca tatagatcca
    10861 agaaacccgc taagaacctc aagatgcctg gcgtctacta tgtggactac agactggaaa
    10921 gaatcaagga ggccaacaac gagacctacg tcgagcagca cgaggtggca gtggccagat
    10981 actgcgacct ccctagcaaa ctggggcaca agcttaatta agaattcgat atcaagctta
    11041 tcggtaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
    11101 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
    11161 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
    11221 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
    11281 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
    11341 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
    11401 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
    11461 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
    11521 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
    11581 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
    11641 accgtcgacc tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta
    11701 ccaatgctga ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca
    11761 cacctcaggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
    11821 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg
    11881 atctgtggat ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc
    11941 cagggatcag atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag
    12001 agaaggtaga agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc
    12061 atgggatgga tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat
    12121 ttcatcacat ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga
    12181 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
    12241 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
    12301 ccctcagacc cttttagtca gtgtggaaaa tctctagcag ggcccgttta aacccgctga
    12361 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
    12421 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
    12481 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
    12541 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
    12601 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
    12661 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
    12721 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
    12781 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
    12841 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
    12901 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
    12961 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
    13021 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
    13081 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
    13141 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
    13201 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
    13261 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
    13321 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
    13381 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
    13441 tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata
    13501 atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac
    13561 cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga
    13621 cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc
    13681 ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga
    13741 cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc
    13801 ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc
    13861 cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga
    13921 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
    13981 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
    14041 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
    14101 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
    14161 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
    14221 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
    14281 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
    14341 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
    14401 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
    14461 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
    14521 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
    14581 aaa
//
