LOCUS       sequence-246639-       10977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes lentivirus expression of L. monocytogenes CRISPR Type I-B 
            Flag-Cas8b2 driven by hUbC promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10977)
  AUTHORS   Pickar-Oliver A, Black JB, Lewis MM, Mutchnick KJ, Klann TS, 
            Gilcrest KA, Sitton MJ, Nelson CE, Barrera A, Bartelt LC, Reddy TE, 
            Beisel CL, Barrangou R, Gersbach CA
  TITLE     Targeted transcriptional modulation with type I CRISPR-Cas systems 
            in human cells.
  JOURNAL   Nat Biotechnol. 2019 Sep 23. pii: 10.1038/s41587-019-0235-7. doi: 
            10.1038/s41587-019-0235-7.
  PUBMED    31548729
REFERENCE   2  (bases 1 to 10977)
  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: "Nat 
            Biotechnol. 2019 Sep 23. pii: 10.1038/s41587-019-0235-7. doi: 
            10.1038/s41587-019-0235-7."
COMMENT     Sequence Label: pLenti-LmoCas8b2
FEATURES             Location/Qualifiers
     source          1..10977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2622..3833
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3670..3689
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      3890..3899
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3899..3922
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3929..3949
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    5669..6257
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5722..5742)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6140..6151)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6260..6276)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6261..6277)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6782..6962
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6988..7005)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6994..7218
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7264..7692
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7351..7370)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7561..7582
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7701..7721)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7706..8035
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7886..8021
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7948..7967
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        8083..8130
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             8149..8523
                     /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    8653..8774
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8690..8709)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8744..8763
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8823..8839)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8823..8839)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8836..8858)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8847..8863
                     /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(8871..8877,8878..8895,8896..8901))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8916..8937
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9054..9071)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9225..9813)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9305..9324)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9984..10775,10776..10844))
                     /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     10607..10626
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10845..10949)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta acccgtgtcg gctccagatc tggcctccgc gccgggtttt
     2641 ggcgcctccc gcgggcgccc ccctcctcac ggcgagcgct gccacgtcag acgaagggcg
     2701 cagcgagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga
     2761 ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc
     2821 tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg
     2881 cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc
     2941 gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga
     3001 tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc
     3061 gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt
     3121 ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt
     3181 cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg
     3241 ggggcatggt gggcggcaag aacccaaggt cttgaggcct tcgctaatgc gggaaagctc
     3301 ttattcgggt gagatgggct ggggcaccat ctggggaccc tgacgtgaag tttgtcactg
     3361 actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt gccgttgggc
     3421 agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt
     3481 tggcttataa tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct tttctccgtc
     3541 gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg ccggacctct
     3601 ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt acctatcttc
     3661 ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg
     3721 aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat tttcagtgtt
     3781 agactagtaa attgtccgct aaattctggc cgtttttggc ttttttgtta gacgaagctt
     3841 gggctgcagg tcgactctag aggatccccg ggtaccggtc gccaccgccg ccaccatgga
     3901 ttacaaggac gatgacgaca aaatggcacc gaagaagaag cggaaggtcc gcggaatgca
     3961 gaccgaaatc gaagtcaggg cagacgactg gctcatcaac accggactga ccggcttcct
     4021 gaacattgtg ggaaaagaga acgtccggat tgacgggcag tctatctact tcaccgctga
     4081 cctcctggaa gatttcgaaa ctaagtactt taacttcttc atccaagagt acaaagaaac
     4141 tctctcatgg tacaagatcg tcagctacaa agagactatg gagcagttca gaattgatga
     4201 attcgcgtcc tttgacgaag tggccctggg gaacctgaac aagtacatga aggacgtcgt
     4261 gaagttctat ctgaaaaagg ccaactatat caaagtgttc cccttgattg accctagcgc
     4321 caacgtgacc aagtggctcg aaaagctgaa caccattaat ctgaccaaga agcagaaatg
     4381 ggatgaggtc aaggctgaca ttattgagga ggtcaaacag acctacaccc aactcgacct
     4441 gattattgac ttctgtgcct ccgaaaaggg actgcagtac ctgggagcca agaacctgat
     4501 ctattccgtc attaacaagg gctggagcgg agtctccttc ctgttcaagc aaaccaagtt
     4561 cattgacccg tatctggact acaagacgac cttcgtggat cctgtgctgg agtacctgga
     4621 taccgatctg tccaaggcca agtacaactg cttcaactgc aaccagccca tcaagaacct
     4681 caagctcgac ctctcgttca tgaatgacgt gggctttgac accgcccgca agactgcgca
     4741 cgtctgggat ttcaacaacg atgtggccac ttgtccaatc tgccggctga tctactcatg
     4801 cgtgcctgcc gggttcacct acgtgtatgg agagggaatg ttcgtgaacg actcctacgg
     4861 cgtggaagaa ctgcacagag tgaacgaacg gatgcggaac agcatcctga ggtttaacaa
     4921 ggacggtatc aattccacca acacttaccg cgcgcttgtg gagtccatta ccatggaaca
     4981 cgaaaacaac agacgctacg aattggctga catccagctc gtgcgatacg agaacgagca
     5041 ttaccggttc aatctgttgt caaagaagat gcttcacatc gtgaacgatt ccaagggtat
     5101 cctgaagtcg ctgatacgct gcggttacaa ggaggggaac ctcaacgtga acctctacaa
     5161 ggaagtgatc cagcatctga tgaacaacga aaacctcttc acccttatcc acaagttgat
     5221 ctactacaag cagaccagcg tgaacggcct gtactacaac atgggccacg tgtccggaat
     5281 cctggatatc aacaccaagt tcctgaaaga aatggaggtc atgacgaaca tctcacagaa
     5341 ccagctgtgg ttcgtgcaaa attgtggcaa agaattcaag gagggttacg tgaagaagaa
     5401 gtccgagaac aagatctccg gaatcactta caagctgctg aattcgctga aggtcaacga
     5461 caaggacgga ttcatggaca ccctgctgaa ctcctactcg tacctgtcca tgccgatccc
     5521 cgacgtgttc atcaagatgt tctccaacaa cgaagccttc aagagcgtgg gctacgcttt
     5581 tatgcttggc gtgggtggag aaaagactaa gaaggaagat gggggcaaca ccgatgagaa
     5641 gtgagaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     5701 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     5761 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     5821 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     5881 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     5941 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     6001 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     6061 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     6121 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     6181 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     6241 ttgggccgcc tccccgcatc gataccgtcg acctcgagac ctagaaaaac atggagcaat
     6301 cacaagtagc aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga
     6361 ggaggaggtg ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc
     6421 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc
     6481 ccaacgaaga caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga
     6541 ttggcagaac tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta
     6601 caagctagta ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg
     6661 cttgttacac cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg
     6721 gaggtttgac agccgcctag catttcatca catggcccga gagctgcatc cggactgtac
     6781 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     6841 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     6901 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     6961 cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     7021 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     7081 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     7141 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     7201 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     7261 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     7321 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     7381 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     7441 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     7501 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     7561 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     7621 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     7681 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     7741 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     7801 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     7861 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     7921 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     7981 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     8041 cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg
     8101 gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg
     8161 accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc
     8221 gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac
     8281 gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc
     8341 tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg
     8401 aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg
     8461 gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac
     8521 tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     8581 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     8641 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     8701 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     8761 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     8821 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     8881 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     8941 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     9001 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     9061 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     9121 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     9181 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     9241 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     9301 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     9361 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     9421 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     9481 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     9541 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     9601 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     9661 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     9721 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     9781 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     9841 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     9901 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     9961 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
    10021 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
    10081 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
    10141 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
    10201 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
    10261 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
    10321 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
    10381 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
    10441 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
    10501 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
    10561 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
    10621 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
    10681 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
    10741 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
    10801 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
    10861 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
    10921 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgac
//
