LOCUS       sequence-449965-       14278 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral Expression of dCas9-KRAB from S. uberis fused to the 
            repressive KRAB domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14278)
  AUTHORS   Butterfield GL, Rohm D, Roberts A, Nethery MA, Rizzo AJ, Morone DJ, 
            Garnier L, Iglesias N, Barrangou R, Gersbach CA
  TITLE     Characterization of diverse Cas9 orthologs for genome and epigenome 
            editing.
  JOURNAL   Proc Natl Acad Sci U S A. 2025 Mar 18;122(11):e2417674122. doi: 
            10.1073/pnas.2417674122. Epub 2025 Mar 12.
  PUBMED    40073054
REFERENCE   2  (bases 1 to 14278)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2025-03-18"; volume: "122(11):e2417674122. 
            doi"; pages: " 10.1073/pnas.2417674122. Epub 2025 Mar 12"
COMMENT     Sequence Label: pFUGW hUbC-Sub-dCas9-KRAB-T2A-GFP
FEATURES             Location/Qualifiers
     source          1..14278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        225..423
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     378..398
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             441..621
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    668..793
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1286..1519
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1704..1748
                     /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    2046..2163
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    complement(2205..2238)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2262..3473
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3310..3329
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      3502..3511
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3511..3576
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3586..3606
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6994..7014
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7051..7236
                     /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             7243..7296
                     /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             join(7303..7305,7306..7308,7309..8019)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7303..7305,7306..7308,7309..8019)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7348..7369)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7609..7628)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7956..7977
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             8029..8085
                     /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"
     protein_bind    complement(8518..8551)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    8576..9164
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8629..8649)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9047..9058)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9167..9183)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9168..9184)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9689..9869
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9895..9912)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9901..10125
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10171..10599
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10258..10277)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10468..10489
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10608..10628)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10613..10942
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10793..10928
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10855..10874
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10990..11037
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11056..11430
                     /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    11560..11681
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11597..11616)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11651..11670
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11730..11746)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11730..11746)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11743..11765)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11754..11770
                     /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(11778..11784,11785..11802,11803..11808))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11823..11844
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11961..11978)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12132..12720)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12212..12231)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12891..13682,13683..13751))
                     /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     13514..13533
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13752..13856)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(13931..13950)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(14122..14278,1..223)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
       61 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      121 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      181 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      241 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      301 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      361 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      421 cagcgcgttt tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct
      481 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
      541 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
      601 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc
      661 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg
      721 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg
      781 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt
      841 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
      901 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact
      961 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
     1021 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
     1081 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga
     1141 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
     1201 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
     1261 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
     1321 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
     1381 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
     1441 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
     1501 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
     1561 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
     1621 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
     1681 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
     1741 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
     1801 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
     1861 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
     1921 ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc
     1981 gattagtgaa cggatcggca ctgcgtgcgc caattctgca gacaaatggc agtattcatc
     2041 cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag aatagtagac
     2101 ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa aattcaaaat
     2161 tttcgggttt attacaggga cagcagagat ccagtttggt taatataact tcgtatagca
     2221 tacattatac gaagttatta acccgtgtcg gctccagatc tggcctccgc gccgggtttt
     2281 ggcgcctccc gcgggcgccc ccctcctcac ggcgagcgct gccacgtcag acgaagggcg
     2341 cagcgagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga
     2401 ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc
     2461 tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg
     2521 cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc
     2581 gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga
     2641 tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc
     2701 gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt
     2761 ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt
     2821 cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg
     2881 ggggcatggt gggcggcaag aacccaaggt cttgaggcct tcgctaatgc gggaaagctc
     2941 ttattcgggt gagatgggct ggggcaccat ctggggaccc tgacgtgaag tttgtcactg
     3001 actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt gccgttgggc
     3061 agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt
     3121 tggcttataa tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct tttctccgtc
     3181 gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg ccggacctct
     3241 ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt acctatcttc
     3301 ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg
     3361 aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat tttcagtgtt
     3421 agactagtaa attgtccgct aaattctggc cgtttttggc ttttttgtta gacgaagctt
     3481 gggctgcagg tcgactctag agccaccatg gactacaaag accatgacgg tgattataaa
     3541 gatcatgaca tcgattacaa ggatgacgat gacaagcacg ttaaccccaa gaagaagagg
     3601 aaggtgggcc gcggaaccgg tatgactaat gggatgattc tgggcttagc aatcggagtc
     3661 gcgtctgtag gggtaggaat tatcgaggcc gatagtggca aggtaattca cgcaagctca
     3721 cggatcttcc ctagtgctaa tgccgacaat aatgtggagc gccgcaagtt ccggggatct
     3781 aggcgccttc ttaggaggaa aaagcacagg gttaagcgct tacaggatct gtttgacaag
     3841 tacgatatcg tgactaactt cgataacctc aaccttaacc cctacgagct gcgagttaag
     3901 ggcttaaacg agccattgag caacgaagag ctcttcgcat cactccggaa catcacaaag
     3961 cacaggggca tttcctatct cgatgatgct gaggatgact cttctggaaa cgggacagaa
     4021 tacgccaaag cgatagagct gaatcagcag cttctgaaag aaaagacccc cggtcagatc
     4081 cagtacgaca gactcaatca gtatgggcaa ctgagaggca atttcgatat cgtggatgaa
     4141 aacggcgaga ttcaccacgt gataaacgtt ttttcaacat caagttacag aaaggaagcc
     4201 gagcagattc tcaagaagca gtctgaaacg aatactagta tcagcaccga ctttataaat
     4261 gatttcatcc aattgctgac ctctaagagg aaatattacc atggtcctgg taatccaaag
     4321 agccgcacag attacgggcg ctaccggacg gatgggacgg atctcgataa catcttcgat
     4381 gttctgatag gtaaatgcag cttttaccca gaggagtacc gagccagcaa gacgagctac
     4441 actgcccaag agttcaactt tcttaatgac ttgaataacc tgaccttacc aaccgagaca
     4501 ggcaagttga gcgagcagca gaagatcgac ctggtgaatt gggctaagga gacaaagatc
     4561 ctcggaccga aaaagctgct tcaggaaatt gccaagagga acaactgcaa gtacgaggac
     4621 attcgcggct atcggcttga taagaaagat aaccccgata tgcatgtatt tgatgtgtat
     4681 cggaagatga attttgacct ggagactatt tccgttaagg atctgtcagt cgactctctg
     4741 aatcagctcg cgcgaattct gacactgaac accgagaggg aggggatcga agaggccatc
     4801 aaaaatctga tgccaaacca gttcaccgag aagcaaatgc ttgaactcat cgccttccgc
     4861 aagagtaatt cctctatctt tgggaagggg tggcacagtc tgtcaattaa actgatgaaa
     4921 gagctgatac ccgagctcta ccacaccagt gacgaacaaa tgaccatact caatcgattt
     4981 ggtaagttca agctcacgaa gctcgactca aaaaggacca attacatcga tgaaaacttt
     5041 gtcactgatg aaatctataa ccctgtagtg gccaagagtg tgaggcaggc aataaagatc
     5101 atcaacgctt ccattaaaaa gtggggggac tttgataaga tcgtgattga gatgccacgc
     5161 gacaagaatg aggaggagga aaggaaacga atcgccgatg gccagaaggt gaatgctaag
     5221 gaaaaagagc aggccgagaa gcacgccgca aagctcttta atggcaagga agagctccct
     5281 tctgaagttt tccatggata taaggagctg gctttgcgaa ttagactctg gtatcagcaa
     5341 gaccagaagt gcctctattc tggcaaggag ataacaattt cagacctgat ctacaacagg
     5401 gagctctttg agattgacgc catccttccg ctgtctcttt cttttgacga cagtctgtct
     5461 aacaaggtcc tggtttacag atgggcaaat caggagaagg gccagaggac ccctttccaa
     5521 gcccttgatt ccatgaaatc agcgtggtcc tatcgggagt tcaagaatgc aatcctgcac
     5581 aattctaaaa tcagccggag aaagcgtgac tattttctga cagaacaaga cattagtaag
     5641 attgaggtga aacaaaagtt tattgagagg aacttggtgg acacacggta cgccagtaga
     5701 acagttctca acgtgctgca gcagtccctg aagaatctgg agaaggagac taaggtgtcc
     5761 gttgtccgag gacagttcac gtcccagctg cgccggaaat ggcacataga taagaccagg
     5821 gatacttacc atcaccatgc ggtggacgca ctgattatcg cggcctccgc taagttgaga
     5881 tattggaaga aacagggcga catcttgttc gagaactatc tcatcaatcg ccacgtagat
     5941 agagtaaccg gggagataca atctgacgat agctataagg aggaggtgtt cacacctccc
     6001 tacgacggat ttgtccagac tattagcaac ccagggtttg aggacgagat ccttttctcc
     6061 tatcaggtag acagtaaagt caacagaaag atctcagacg ccacgatata cgctacgagg
     6121 tctgcgaagc tcgagaagga caagaaggaa cagacgtatg tcttgggtaa gataaaagat
     6181 atctattcac aaactggttt tgagaacttc ctgaagatct ataataagga caagagtaag
     6241 ttcctgatct accagaagga ccctgagact tgggaaaaga tcattgaacc aattctcaaa
     6301 aattatcggg aattcgataa taaaggcaag gatatcgtga atccatttga gaaatacagg
     6361 aatgataacg ggcctatctg caagtacagt cggaaaggca acggccctga gatcaaacaa
     6421 tttaaatact acgacaccgt ttacaaaatt acaagcggtc tcgacatcag cccccgcgaa
     6481 tcaagaaata aggtaattct tcaaagcctg aatccgtgga gaaccgactt ctactttaac
     6541 cctaagacta tgaagtacga acttatgggt atcagatatg tcgacctgga gttcgagaaa
     6601 ggaacagggg actacctgat ttctgacaat ctctataaag agattaaaaa gaacgagggg
     6661 atctctgagc tgagtgtatt caagttcaca ctctacaaga acgatctcct gctgatcaag
     6721 gacactgaga acaacgaaga gcaaattttt aggttttggt ctcggaatga cctgtcctcc
     6781 aaaaaccggg tggaactgaa gccctacgat aggtcccgct tttccggcaa tgagatcctt
     6841 atcaccaaaa tgggcaaggc acctaagcaa tgcattaaga ctttaacata ccaaaacatc
     6901 tccatttata aaatcaaaac agacatcctg ggattcaaat actatctgaa aaacgaagga
     6961 aataagccat tactgcactt taagaagacc ggtcctaaga aaaagcggaa agtggctagc
     7021 atggatgcta agtcactaac tgcctggtcc cggacactgg tgaccttcaa ggatgtattt
     7081 gtggacttca ccagggagga gtggaagctg ctggacactg ctcagcagat cgtgtacaga
     7141 aatgtgatgc tggagaacta taagaacctg gtttccttgg gttatcagct tactaagcca
     7201 gatgtgatcc tccggttgga gaagggagaa gagcccgcta gcgagggcag aggaagtctt
     7261 ctaacatgcg gtgacgtgga ggagaatccc ggccctggac caatggtgag caagggcgag
     7321 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     7381 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     7441 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     7501 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     7561 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     7621 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     7681 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     7741 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     7801 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     7861 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     7921 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     7981 gccgccggga tcactctcgg catggacgag ctgtacaagg gatccggcgc aacaaacttc
     8041 tctctgctga aacaagccgg agatgtcgaa gagaatcctg gaccgaagac cttcaacatc
     8101 tctcagcagg atctggagct ggtggaggtc gccactgaga agatcaccat gctctatgag
     8161 gacaacaagc accatgtcgg ggcggccatc aggaccaaga ctggggagat catctctgct
     8221 gtccacattg aggcctacat tggcagggtc actgtctgtg ctgaagccat tgccattggg
     8281 tctgctgtga gcaacgggca gaaggacttt gacaccattg tggctgtcag gcacccctac
     8341 tctgatgagg tggacagatc catcagggtg gtcagcccct gtggcatgtg cagagagctc
     8401 atctctgact atgctcctga ctgctttgtg ctcattgaga tgaatggcaa gctggtcaaa
     8461 accaccattg aggaactcat ccccctcaag tacaccagga actaaagttg ataatagata
     8521 acttcgtata gcatacatta tacgaagtta tgaattcgat atcaagctta tcgataatca
     8581 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     8641 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     8701 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     8761 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     8821 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     8881 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     8941 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     9001 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     9061 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     9121 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc
     9181 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
     9241 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
     9301 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     9361 ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
     9421 ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc cagggatcag
     9481 atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga
     9541 agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga
     9601 tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat
     9661 ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg
     9721 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     9781 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     9841 cttttagtca gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga
     9901 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     9961 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
    10021 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
    10081 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
    10141 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
    10201 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
    10261 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
    10321 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
    10381 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
    10441 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
    10501 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
    10561 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
    10621 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
    10681 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
    10741 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
    10801 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
    10861 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
    10921 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
    10981 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
    11041 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
    11101 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
    11161 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
    11221 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
    11281 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
    11341 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
    11401 gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc
    11461 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
    11521 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
    11581 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
    11641 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
    11701 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
    11761 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
    11821 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
    11881 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
    11941 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
    12001 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
    12061 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
    12121 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
    12181 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
    12241 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
    12301 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
    12361 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
    12421 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
    12481 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
    12541 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
    12601 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
    12661 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
    12721 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
    12781 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
    12841 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
    12901 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
    12961 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
    13021 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
    13081 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
    13141 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
    13201 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
    13261 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
    13321 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
    13381 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
    13441 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
    13501 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
    13561 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
    13621 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
    13681 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
    13741 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
    13801 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
    13861 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
    13921 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
    13981 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
    14041 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
    14101 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
    14161 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
    14221 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgt
//
