LOCUS       sequence-377036-       11821 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral expression of S. aureus dead Cas9 
            (dCas9/dSaCas9/SadCas9), eGFP, and the blasticidin resistance gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11821)
  AUTHORS   Sapozhnikov DM, Szyf M
  TITLE     The PROTECTOR strategy employs dCas orthologs to sterically shield 
            off-target sites from CRISPR/Cas activity.
  JOURNAL   Sci Rep. 2023 Feb 9;13(1):2280. doi: 10.1038/s41598-023-29332-2.
  PUBMED    36759683
REFERENCE   2  (bases 1 to 11821)
  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: "Sci Rep."; 
            date: "2023-02-9"; volume: "13(1):2280. doi"; pages: " 
            10.1038/s41598-023-29332-2"
COMMENT     Sequence Label: pLenti-eGFP-dSaCas9-blast
FEATURES             Location/Qualifiers
     source          1..11821
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     13..30
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    147..168
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(183..188,189..206,207..213)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    221..237
                     /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     226..248
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     245..261
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     245..261
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     280..300
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        282..300
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        328..554
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             555..735
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    782..907
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1400..1633
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1818..1862
                     /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    2129..2246
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2269..2572
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2573..2776
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2727..2747
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2773..2797
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2876..2885
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2882..2884,2885..2887,2888..3598)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2882..2884,2885..2887,2888..3598)
                     /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(2927..2948)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3188..3207)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3535..3556
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3608..3664
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3686..3739
                     /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             6914..6934
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6941..7006
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7084..7125
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    7149..7737
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7202..7222)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7620..7631)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        7752..8068
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7919..8054
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        8116..8163
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8182..8580
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             8667..8900
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8972..9093
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9009..9028)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9063..9082
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9133..9268
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9288..9307)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9289..9307)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9317..9334)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9317..9333)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9326..9348)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9475..9930
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9562..9581)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9772..9793
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9956..10060
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10061..10129,10130..10921)
                     /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(10279..10298)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11092..11680
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11581..11600
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
       61 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
      121 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
      181 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
      241 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
      301 gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca
      361 acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc
      421 gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga
      481 catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag
      541 ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct
      601 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
      661 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
      721 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
      781 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
      841 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
      901 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
      961 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
     1021 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
     1081 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
     1141 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
     1201 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca
     1261 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
     1321 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
     1381 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
     1441 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
     1501 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
     1561 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
     1621 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
     1681 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
     1741 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat
     1801 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt
     1861 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag
     1921 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aatagagtta
     1981 ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg ggacccgaca
     2041 ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc attcgattag
     2101 tgaacggatc tcgacggtat cggttaactt ttaaaagaaa aggggggatt ggggggtaca
     2161 gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa
     2221 aacaaattac aaaaattcaa aattttatcg ataagcttgg gagttccgcg ttacataact
     2281 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     2341 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     2401 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     2461 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     2521 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     2581 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     2641 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     2701 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     2761 ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg
     2821 ttttgacctc catagaagac accgactcta gggctgcagg tccgatccac cggtcgccac
     2881 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     2941 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     3001 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     3061 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     3121 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     3181 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     3241 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     3301 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     3361 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     3421 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     3481 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     3541 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg
     3601 atctggagca acaaacttct cactactcaa acaagcaggt gacgtggagg agaatcccgg
     3661 gccttctaga ggatccggct ccggcgaggg caggggaagt ctactaacat gcggggacgt
     3721 ggaggaaaat cccggcccag ctagcatggg caaacggaac tacatcctgg ggcttgccat
     3781 tgggataacc agcgttggct acggaattat tgattatgag acacgcgatg tgattgacgc
     3841 cggggttagg ctgttcaaag aggccaacgt tgaaaacaac gagggaagac ggagtaagcg
     3901 cggagcaaga agactcaagc gcagacggag acatcggatt cagagggtga aaaagctgct
     3961 cttcgattac aatctcctga ccgatcatag tgagctgagc ggaatcaacc cctacgaggc
     4021 gcgagtgaaa gggctttccc agaagctgtc cgaagaggag ttctccgccg cgttgctgca
     4081 cctggccaaa cggagggggg ttcacaatgt aaacgaagtg gaggaggaca cgggcaatga
     4141 acttagtacg aaagaacaga tcagtaggaa ctctaaggct ctcgaagaga aatacgtcgc
     4201 tgagttgcag cttgagagac tgaaaaaaga cggcgaagta cgcggatcta ttaataggtt
     4261 caagacttca gattacgtaa aggaagccaa gcagctcctg aaagtacaga aagcgtacca
     4321 tcagctcgat cagagcttca tcgataccta catagatttg ctggagacac ggaggacata
     4381 ctacgagggc ccaggggaag gatctccttt tgggtggaag gacatcaagg aatggtacga
     4441 gatgcttatg ggacattgta catattttcc ggaggagctc aggagcgtca agtacgccta
     4501 caatgccgac ctgtacaatg ccctcaatga cctcaataac ctcgtgatta ccagggacga
     4561 gaacgagaag ctggagtact atgaaaagtt ccagattatc gagaatgtgt ttaagcagaa
     4621 gaagaagccg acacttaagc agattgcaaa ggaaatcctc gtgaatgagg aagatatcaa
     4681 gggatacaga gtgacaagta caggcaagcc cgagttcaca aatctgaagg tgtaccacga
     4741 tattaaggac ataaccgcac gaaaggagat aatcgaaaac gctgagctcc tcgatcagat
     4801 cgcaaaaatt cttaccatct accagtctag tgaggacatt caggaggaac tgactaatct
     4861 gaacagtgag ctcacccaag aggaaattga gcagatttca aacctgaaag gctacaccgg
     4921 gacgcacaat ctgagcctca aagcaatcaa cctcattctg gatgaacttt ggcacacaaa
     4981 tgacaaccaa attgccatat tcaaccgcct gaaactggtg ccaaaaaaag tggatctgtc
     5041 acagcaaaag gaaatcccta caaccttggt tgacgatttt attctgtccc ccgttgtcaa
     5101 gcggagcttc atccagtcaa tcaaggtgat caatgccatc attaaaaaat acggattgcc
     5161 aaacgatata attatcgagc ttgcacgaga gaagaactca aaggacgccc agaagatgat
     5221 taacgaaatg cagaagcgca accgccagac aaacgaacgc atagaggaaa ttataagaac
     5281 aaccggcaaa gagaatgcca agtatctgat cgagaaaatc aagctgcacg acatgcaaga
     5341 aggcaagtgc ctgtactctc tggaagctat cccactcgaa gatctgctga ataatccatt
     5401 caattacgag gtggacgcca tcatccctag atccgtaagc tttgacaatt ccttcaataa
     5461 caaagttctg gttaaacagg aggaaaattc taaaaaaggg aaccggaccc cgttccagta
     5521 cctgagctcc agtgacagca agattagcta cgagactttt aagaaacata ttctgaatct
     5581 ggccaaaggc aaaggcagga tcagcaagac caagaaggag tacctcctcg aagaacgcga
     5641 cattaacaga tttagtgtgc agaaagattt catcaaccga aaccttgtcg atactcggta
     5701 cgccacgaga ggcctgatga atctcctcag gagctacttc cgcgtcaata atctggacgt
     5761 taaagtcaag agcataaatg ggggattcac cagctttctg aggagaaagt ggaagtttaa
     5821 gaaggaacga aacaaaggat acaagcacca tgctgaggat gctttgatca tcgctaacgc
     5881 ggactttatc tttaaggaat ggaaaaagct ggataaggca aagaaagtga tggaaaacca
     5941 gatgttcgag gagaagcagg cagagtcaat gcctgagatc gagacagagc aggaatacaa
     6001 ggaaattttc atcacccctc atcagattaa acacataaag gacttcaaag actataaata
     6061 ctctcatagg gtggacaaaa aacccaatcg cgagctcatt aatgacaccc tgtactcaac
     6121 acggaaggat gataaaggta ataccttgat tgtgaataat cttaatggat tgtatgacaa
     6181 agataacgac aagctcaaga agctgatcaa caagtctcca gagaagctcc ttatgtatca
     6241 ccacgaccca cagacttatc agaaattgaa actgatcatg gagcaatacg gggatgagaa
     6301 gaacccactc tacaaatatt atgaggaaac aggtaattac ctgaccaagt actccaagaa
     6361 ggataacgga ccagtgatca aaaagataaa gtactatggc aacaaactta atgcgcattt
     6421 ggacataact gacgattacc ccaattctcg aaacaaggtt gtgaagctct ccctgaagcc
     6481 ttatagattt gacgtgtacc tggataatgg ggtttataaa ttcgtcaccg tgaaaaatct
     6541 ggacgtgatc aaaaaggaga actattatga agtaaactca aagtgctatg aggaggcgaa
     6601 gaagctgaag aagatctcca atcaggccga gttcatcgct tccttctata ataacgatct
     6661 catcaagatc aatggagagc tttatcgcgt cattggtgtg aacaatgact tgctgaacag
     6721 gatcgaagtc aatatgatag acattaccta ccgggagtat ctcgaaaaca tgaatgataa
     6781 acggccgcct cgcatcatca agacaatcgc atctaaaact cagtcaataa aaaagtactc
     6841 taccgatatc ctggggaatc tctatgaagt gaagtcaaag aagcacccac aaatcattaa
     6901 aaaaggtgga tcccccaaga agaagaggaa agtctcgagc gactacaaag accatgacgg
     6961 tgattataaa gatcatgaca tcgattacaa ggatgacgat gacaagtaaa gcggccgcca
     7021 ctgtgctgga tatctgcaga attcgttccg gagtcgtcga ctctagaggg cccgcggttc
     7081 gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggttagtaa
     7141 tgatcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     7201 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     7261 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     7321 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     7381 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     7441 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     7501 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat
     7561 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     7621 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     7681 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
     7741 gcgatggtac cggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     7801 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     7861 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     7921 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     7981 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     8041 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     8101 atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac
     8161 gacaaggtga ggaactaaac catggccaag cctttgtctc aagaagaatc caccctcatt
     8221 gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag cgtcgccagc
     8281 gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca ttttactggg
     8341 ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc tggcaacctg
     8401 acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg cggacggtgc
     8461 cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga cagtgatgga
     8521 cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa
     8581 gcacaattcg agctcggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     8641 ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga
     8701 tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     8761 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     8821 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     8881 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact
     8941 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     9001 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     9061 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     9121 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     9181 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     9241 aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt
     9301 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     9361 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     9421 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     9481 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     9541 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     9601 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     9661 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     9721 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     9781 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     9841 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     9901 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     9961 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
    10021 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
    10081 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
    10141 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
    10201 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
    10261 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
    10321 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
    10381 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
    10441 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
    10501 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
    10561 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
    10621 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
    10681 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
    10741 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
    10801 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
    10861 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
    10921 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
    10981 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
    11041 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
    11101 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
    11161 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
    11221 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
    11281 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
    11341 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
    11401 gtcgggctga acggggggtt cgtgctcaca gcccagcttg gagcgaacga cctacaccga
    11461 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
    11521 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
    11581 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
    11641 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
    11701 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
    11761 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
    11821 a
//
