LOCUS       sequence-338623-       12435 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Lentiviral dSpCas9-KRAB (CRISPRi) delivery, mNeon coexpression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12435)
  TITLE     Lentiviral CRISPR/Cas9 plasmids with mNeon
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 12435)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: SIN40C.SFFV.dCas9-KRAB.P2A.mNeon
FEATURES             Location/Qualifiers
     source          1..12435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    95..116
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(131..136,137..154,155..161)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    169..185
                     /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     174..196
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     193..209
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     193..209
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     228..248
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        230..248
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     enhancer        293..672
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     enhancer        729..920
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(752..772)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        939..1165
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1166..1346
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1393..1518
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2011..2244
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2429..2473
                     /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    2686..2803
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2861..3268
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      3268..3277
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3280..3306
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3310..3330
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3340..3363,3364..3366,3367..5922,5923..5925,
                     5926..7440)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(3340..3363,3364..3366,3367..5853,5854..5856,
                     5857..7440)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             7441..7488
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7501..7521
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7570..7755
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     CDS             7780..7836
                     /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"
     regulatory      8092..8101
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    8563..9151
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8616..8636)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9034..9045)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9223..9456
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9534..9655
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9571..9590)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9625..9644
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9695..9830
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9850..9869)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9851..9869)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9879..9896)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9879..9895)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9888..9910)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      10037..10492
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10124..10143)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10334..10355
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10518..10622
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10623..10691,10692..11483)
                     /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(10841..10860)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11654..12242
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12143..12162
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12396..12413
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
       61 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      121 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      181 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
      241 actaaaggga acaaaagctg gagctgcaag cggtaccgtc gacctgcagg ttgacattga
      301 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      361 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      421 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      481 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      541 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      601 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      661 gctattacca tggcgcgcca tgcagggcct gaaataacct ctgaaagagg aacttggtta
      721 ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
      781 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
      841 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
      901 tagtcagcaa ccatagtccc ttaagaggcg cgccttaatg tagtcttatg caatactctt
      961 gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca
     1021 ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca
     1081 gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt
     1141 taagtgccta gctcgataca taaacgggtc tctctggtta gaccagatct gagcctggga
     1201 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     1261 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     1321 ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg
     1381 aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc
     1441 gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga
     1501 gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt
     1561 cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg
     1621 gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa
     1681 atactgggac agctacaacc atcccttcag acaggatcag aagaacttag atcattatat
     1741 aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa
     1801 gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc
     1861 gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg aattatataa
     1921 atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt
     1981 ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc
     2041 agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt
     2101 gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct
     2161 gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag
     2221 atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac
     2281 cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca
     2341 cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt
     2401 aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa
     2461 atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt
     2521 cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt
     2581 gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag
     2641 gggacaattc tagatcttga gacaaatggc agtattcatc cacaatttta aaagaaaagg
     2701 ggggattggg gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca
     2761 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga
     2821 cagcagagat ccactttggc gccggctcga gcgagctgca gtaacgccat tttgcaaggc
     2881 atggaaaaat accaaaccaa gaatagagaa gttcagatca agggcgggta catgaaaata
     2941 gctaacgttg ggccaaacag gatatctgcg gtgagcagtt tcggccccgg cccggggcca
     3001 agaacagatg gtcaccgcag tttcggcccc ggcccgaggc caagaacaga tggtccccag
     3061 atatggccca accctcagca gtttcttaag acccatcaga tgtttccagg ctcccccaag
     3121 gacctgaaat gaccctgcgc cttatttgaa ttaaccaatc agcctgcttc tcgcttctgt
     3181 tcgcgcgctt ctgcttcccg agctctataa aagagctcac aacccctcac tcggcgcgcc
     3241 agtcctccga cagactgagt cggatccgcc accatgggct acccatacga tgttccagat
     3301 tacgcttcgc cgaagaaaaa gcgcaaggtc gaagcgtccg acaagaagta cagcatcggc
     3361 ctggccatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc
     3421 agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc
     3481 ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc
     3541 agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac
     3601 gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa
     3661 gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac
     3721 cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag
     3781 gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc
     3841 ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg
     3901 gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc
     3961 aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag
     4021 ctgcccggcg agaagaagaa tggcctgttc ggcaacctga ttgccctgag cctgggcctg
     4081 acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag
     4141 gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac
     4201 ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg
     4261 aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac
     4321 caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa
     4381 gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc
     4441 caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa
     4501 ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc
     4561 agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat
     4621 ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc
     4681 ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag
     4741 agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc
     4801 cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg
     4861 cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa
     4921 tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc
     4981 gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac
     5041 ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac
     5101 gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac
     5161 aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac
     5221 agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg
     5281 aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac
     5341 ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc
     5401 gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc
     5461 cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc
     5521 ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg
     5581 aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag
     5641 accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc
     5701 aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac
     5761 gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg
     5821 gacatcaacc ggctgtccga ctacgatgtg gacgccatcg tgcctcagag ctttctgaag
     5881 gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac
     5941 aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac
     6001 gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg
     6061 agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca
     6121 aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag
     6181 ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag
     6241 gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac
     6301 ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc
     6361 gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa
     6421 atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc
     6481 gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa
     6541 accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc
     6601 atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag
     6661 tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct
     6721 aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtactggt ggtggccaaa
     6781 gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc
     6841 atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa
     6901 gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac
     6961 ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg
     7021 ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc
     7081 cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag
     7141 atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac
     7201 aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat
     7261 atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac
     7321 accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc
     7381 caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac
     7441 aagcgtcctg ctgctactaa gaaagctggt caagctaaga aaaagaaagc tagaggctct
     7501 cccaagaaaa agagaaaggt tgaaggcgga ggcggtggca tggatgctaa gtcactaact
     7561 gcctggtccc ggacactggt gaccttcaag gatgtatttg tggacttcac cagggaggag
     7621 tggaagctgc tggacactgc tcagcagatc gtgtacagaa atgtgatgct ggagaactat
     7681 aagaacctgg tttccttggg ttatcagctt actaagccag atgtgatcct ccggttggag
     7741 aagggagaag agcccggcgg agctagcgga ggaagcggcg ccaccaattt cagcctgctg
     7801 aaacaggccg gcgacgtgga agagaaccct ggccctatgg tgtccaaggg cgaagaggac
     7861 aacatggcca gcctgcctgc cacccacgag ctgcacatct tcggcagcat caacggcgtg
     7921 gacttcgaca tggtgggaca gggcaccggc aaccccaacg acggctacga ggaactgaac
     7981 ctgaagtcca caaagggcga cctgcagttc agcccctgga ttctggtgcc ccacatcggc
     8041 tacggcttcc accagtacct gccctacccc gacggcatga gccctttcca agccgccatg
     8101 gtggatggca gcggctacca ggtgcaccgg accatgcagt ttgaggacgg cgccagcctg
     8161 accgtgaact acaggtacac atacgagggc agccacatca agggcgaggc ccaagtgaag
     8221 ggcacaggct ttccagccga cggccccgtg atgaccaata gcctgacagc cgccgactgg
     8281 tgcagaagca agaaaaccta ccccaatgac aagaccatca tcagcacctt caagtggtcc
     8341 tacaccaccg gcaatggcaa gcggtacaga agcaccgccc ggaccaccta caccttcgcc
     8401 aaacccatgg ccgccaacta cctgaagaac cagcctatgt acgtgttccg caagaccgag
     8461 ctgaagcact ccaagacaga actgaacttc aaagagtggc agaaagcctt caccgacgtg
     8521 atgggcatgg acgagctgta caagtgaacg cgttaagtcg acaatcaacc tctggattac
     8581 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     8641 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     8701 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     8761 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     8821 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     8881 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     8941 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     9001 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     9061 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     9121 agtcggatct ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg
     9181 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     9241 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     9301 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     9361 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     9421 tcagaccctt ttagtcagtg tggaaaatct ctagcagtac gtatagtagt tcatgtcatc
     9481 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     9541 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     9601 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     9661 tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     9721 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     9781 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta
     9841 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     9901 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     9961 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
    10021 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
    10081 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
    10141 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
    10201 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
    10261 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
    10321 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
    10381 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
    10441 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
    10501 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
    10561 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
    10621 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
    10681 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
    10741 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
    10801 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
    10861 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
    10921 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
    10981 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
    11041 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
    11101 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
    11161 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
    11221 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
    11281 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
    11341 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
    11401 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
    11461 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
    11521 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
    11581 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
    11641 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
    11701 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
    11761 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
    11821 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
    11881 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
    11941 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
    12001 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
    12061 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
    12121 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
    12181 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
    12241 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
    12301 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
    12361 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
    12421 aagagcgccc aatac
//
