LOCUS       sequence-255304-       12864 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Tet-regulated (Tet-off) lentiviral vector for transgene (CAG 
            promoter) - OR - shRNA (H1 promoter when subcloned from pLVTHM 
            (Addgene#12247)) - 2nd generation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12864)
  AUTHORS   Szulc J, Wiznerowicz M, Sauvain MO, Trono D, Aebischer P
  TITLE     A versatile tool for conditional gene expression and knockdown.
  JOURNAL   Nat Methods. 2006 Feb;3(2):109-16. doi: 10.1038/nmeth846
  PUBMED    16432520
REFERENCE   2  (bases 1 to 12864)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nat Methods"; date: "2006-02"; volume: "3"; issue: "2"; pages: 
            "109-16"
COMMENT     Sequence Label: pLVCT-rtTR-KRAB-2SM2
FEATURES             Location/Qualifiers
     source          1..12864
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             6..639
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    686..811
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1304..1537
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1722..1766
                     /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"
     enhancer        2061..2440
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2442..2717
                     /label=chicken beta-actin promoter
     intron          2718..3734
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3658..3680
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3742..3761
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    3820..3937
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      4014..4023
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4020..4022,4023..4025,4026..4736)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4020..4022,4023..4025,4026..4736)
                     /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(4065..4086)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4326..4345)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4673..4694
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4797..5364
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4815..5366
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4959..4976)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5186..5205
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5990..6010
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6113..6307
                     /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
                     QLTKPDVILRLEKGEEPWLV"
     misc_feature    6449..7037
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6502..6522)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6920..6931)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    join(7175..7193,7194..7216,7217..7235,7236..7258,7259..7277,
                     7278..7300,7301..7319,7320..7342,7343..7361,7362..7384,
                     7385..7403,7404..7426,7427..7445)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     LTR             7484..7664
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(7689..7707)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7690..7707)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(7997..8016)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(8230..8249)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8349..8371
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8409..8427)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8495..8599
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8600..8668,8669..9460)
                     /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(8818..8837)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9631..10219
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10120..10139
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10373..10390
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(10460..10480)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10465..10794
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10645..10780
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          11928..11993
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             12123..12143
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    12568..12702
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12605..12624)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12659..12678
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 gtagttggaa gggctaattc actcccaaag aagacaagat atccttgatc tgtggatcta
       61 ccacacacaa ggctacttcc ctgattagca gaactacaca ccagggccag gggtcagata
      121 tccactgacc tttggatggt gctacaagct agtaccagtt gagccagata aggtagaaga
      181 ggccaataaa ggagagaaca ccagcttgtt acaccctgtg agcctgcatg ggatggatga
      241 cccggagaga gaagtgttag agtggaggtt tgacagccgc ctagcatttc atcacgtggc
      301 ccgagagctg catccggagt acttcaagaa ctgctgatat cgagcttgct acaagggact
      361 ttccgctggg gactttccag ggaggcgtgg cctgggcggg actggggagt ggcgagccct
      421 cagatcctgc atataagcag ctgctttttg cctgtactgg gtctctctgg ttagaccaga
      481 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
      541 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
      601 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
      661 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
      721 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
      781 ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa ttagatcgcg
      841 atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa aacatatagt
      901 atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag aaacatcaga
      961 aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat cagaagaact
     1021 tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga tagagataaa
     1081 agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta agaccaccgc
     1141 acagcaagcg gccgctgatc ttcagacctg gaggaggaga tatgagggac aattggagaa
     1201 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     1261 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     1321 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
     1381 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     1441 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     1501 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     1561 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     1621 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
     1681 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     1741 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     1801 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     1861 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     1921 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
     1981 acagagacag atccattcga ttagtgaacg gatctcgacg gtatcgatca cgagactagc
     2041 ctcgaccatc gatgtcgatc gacattgatt attgactagt tattaatagt aatcaattac
     2101 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     2161 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     2221 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     2281 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     2341 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     2401 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt
     2461 ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt
     2521 ttaattattt tgtgcagcga tgggggcggg gggggggggg gggcgcgccr ggsggggggg
     2581 ggsggggggg gggggggggs ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg
     2641 gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc
     2701 gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg
     2761 ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc
     2821 gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt
     2881 ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg
     2941 gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc
     3001 ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg
     3061 aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag
     3121 gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc
     3181 tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct tcgggtgcgg
     3241 ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg
     3301 gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg
     3361 gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt
     3421 aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg
     3481 ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga
     3541 aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc
     3601 agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt
     3661 tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt catgccttct
     3721 tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat cattttggca
     3781 aagaattaaa tttaattaat ctcgacggta tcggttaact tttaaaagaa aaggggggat
     3841 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     3901 agaattacaa aaacaaatta caaaaattca aaattttatc gatcacgaga ctagcctcga
     3961 ggtttaaact acgggatcca ggcctaagct tacgcgtcct agcgctaccg gtcgccacca
     4021 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     4081 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     4141 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     4201 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     4261 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     4321 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     4381 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     4441 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     4501 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     4561 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     4621 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     4681 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtccg
     4741 gactcagatc tcgactagct agtagctagc tagctagtcg agctcaagct tcgaattccg
     4801 cccccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
     4861 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
     4921 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
     4981 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc
     5041 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     5101 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     5161 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
     5221 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt
     5281 tacatgtgtt tagtcgaggt taaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt
     5341 ttcctttgaa aaacacgatg ataataccat ggctagactg gacaagagca aagtcataaa
     5401 cggcgctctg gaattactca atggagtcgg tatcgaaggc ctgacgacaa ggaaactcgc
     5461 tcaaaagctg ggagttgagc agcctaccct gtactggcac gtgaagaaca agcgggccct
     5521 gctcgatgcc ctgccaatcg agatgctgga caggcatcat acccacttct gccccctgga
     5581 aggcgagtca tggcaagact ttctgcggaa caacgccaag tcattccgct gtgctctcct
     5641 ctcacatcgc gacggggcta aagtgcatct cggcacccgc ccaacagaga aacagtacga
     5701 aaccctggaa aatcagctcg cgttcctgtg tcagcaaggc ttctccctgg agaacgcact
     5761 gtacgctctg tccgccgtgg gccactttac actgggctgc gtattggagg aacaggagca
     5821 tcaagtagca aaagaggaaa gagagacacc taccaccgat tctatgcccc cacttctgag
     5881 acaagcaatt gagctgttcg accggcaggg agccgaacct gccttccttt tcggcctgga
     5941 actaatcata tgcggattag aaaaacaact taaatgtgaa agtgggtcgc caaaaaagaa
     6001 gagaaaggtc gacggcggtg gtgctttgtc tcctcagcac tctgctgtca ctcaaggaag
     6061 tatcatcaag aacaaggagg gcatggatgc taagtcacta actgcctggt cccggacact
     6121 ggtgaccttc aaggatgtat ttgtggactt caccagggag gagtggaagc tgctggacac
     6181 tgctcagcag atcgtgtaca gaaatgtgat gctggagaac tataagaacc tggtttcctt
     6241 gggttatcag cttactaagc cagatgtgat cctccggttg gagaagggag aagagccctg
     6301 gctggtggag agagaaattc accaagagac ccatcctgat tcagagactg catttgaaat
     6361 caaatcatca gtttaaggat ctcgactagc tagtagctag ctagctagtc gagctcaagc
     6421 ttcgaattcc cgggactagt catatgataa tcaacctctg gattacaaaa tttgtgaaag
     6481 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     6541 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     6601 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     6661 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     6721 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     6781 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     6841 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     6901 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     6961 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     7021 ttgggccgcc tccccgcatc ggtacgtatg gccaggtacc tttaagacca atgacttaca
     7081 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     7141 actcccaacg aagacaagat ctcgagttta ccactcccta tcagtgatag agaaaagtga
     7201 aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc
     7261 cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa
     7321 aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta
     7381 ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat
     7441 agagaaaagt gaaagtcgga tcgatctgct ttttgcttgt actgggtctc tctggttaga
     7501 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     7561 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     7621 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagcatct agaattaatt
     7681 ccgtgtattc tatagtgtca cctaaatcgt atgtgtatga tacataaggt tatgtattaa
     7741 ttgtagccgc gttctaacga caatatgtac aagcctaatt gtgtagcatc tggcttactg
     7801 aagcagaccc tatcatctct ctcgtaaact gccgtcagag tcggtttggt tggacgaacc
     7861 ttctgagttt ctggtaacgc cgtcccgcac ccggaaatgg tcagcgaacc aatcagcagg
     7921 gtcatcgcta gccagatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca
     7981 ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac
     8041 ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga
     8101 ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc
     8161 aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgaatggtg
     8221 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     8281 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     8341 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     8401 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     8461 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     8521 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     8581 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     8641 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     8701 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     8761 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     8821 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     8881 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     8941 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     9001 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     9061 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     9121 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     9181 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     9241 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     9301 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     9361 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     9421 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     9481 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     9541 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     9601 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     9661 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     9721 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     9781 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     9841 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     9901 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     9961 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
    10021 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
    10081 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
    10141 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
    10201 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
    10261 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
    10321 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
    10381 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
    10441 gattcattaa tgcagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
    10501 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
    10561 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
    10621 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
    10681 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
    10741 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt
    10801 ggacacaaga caggcttgcg agatatgttt gagaatacca ctttatcccg cgtcagggag
    10861 aggcagtgcg taaaaagacg cggactcatg tgaaatactg gtttttagtg cgccagatct
    10921 ctataatctc gcgcaaccta ttttcccctc gaacactttt taagccgtag ataaacaggc
    10981 tgggacactt cacatgagcg aaaaatacat cgtcacctgg gacatgttgc agatccatgc
    11041 acgtaaactc gcaagccgac tgatgccttc tgaacaatgg aaaggcatta ttgccgtaag
    11101 ccgtggcggt ctgtaccggg tgcgttactg gcgcgtgaac tgggtattcg tcatgtcgat
    11161 accgtttgta tttccagcta cgatcacgac aaccagcgcg agcttaaagt gctgaaacgc
    11221 gcagaaggcg atggcgaagg cttcatcgtt attgatgacc tggtggatac cggtggtact
    11281 gcggttgcga ttcgtgaaat gtatccaaaa gcgcactttg tcaccatctt cgcaaaaccg
    11341 gctggtcgtc cgctggttga tgactatgtt gttgatatcc cgcaagatac ctggattgaa
    11401 cagccgtggg atatgggcgt cgtattcgtc ccgccaatct ccggtcgcta atcttttcaa
    11461 cgcctggcac tgccgggcgt tgttcttttt aacttcaggc gggttacaat agtttccagt
    11521 aagtattctg gaggctgcat ccatgacaca ggcaaacctg agcgaaaccc tgttcaaacc
    11581 ccgctttaaa catcctgaaa cctcgacgct agtccgccgc tttaatcacg gcgcacaacc
    11641 gcctgtgcag tcggcccttg atggtaaaac catccctcac tggtatcgca tgattaaccg
    11701 tctgatgtgg atctggcgcg gcattgaccc acgcgaaatc ctcgacgtcc aggcacgtat
    11761 tgtgatgagc gatgccgaac gtaccgacga tgatttatac gatacggtga ttggctaccg
    11821 tggcggcaac tggatttatg agtgggcccc ggatctttgt gaaggaacct tacttctgtg
    11881 gtgtgacata attggacaaa ctacctacag agatttaaag ctctaaggta aatataaaat
    11941 ttttaagtgt ataatgtgtt aaactactga ttctaattgt ttgtgtattt tagattccaa
    12001 cctatggaac tgatgaatgg gagcagtggt ggaatgcctt taatgaggaa aacctgtttt
    12061 gctcagaaga aatgccatct agtgatgatg aggctactgc tgactctcaa cattctactc
    12121 ctccaaaaaa gaagagaaag gtagaagacc ccaaggactt tccttcagaa ttgctaagtt
    12181 ttttgagtca tgctgtgttt agtaatagaa ctcttgcttg ctttgctatt tacaccacaa
    12241 aggaaaaagc tgcactgcta tacaagaaaa ttatggaaaa atattctgta acctttataa
    12301 gtaggcataa cagttataat cataacatac tgttttttct tactccacac aggcatagag
    12361 tgtctgctat taataactat gctcaaaaat tgtgtacctt tagcttttta atttgtaaag
    12421 gggttaataa ggaatatttg atgtatagtg ccttgactag agatcataat cagccatacc
    12481 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
    12541 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
    12601 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
    12661 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcaactggat aactcaagct
    12721 aaccaaaatc atcccaaact tcccacccca taccctatta ccactgccaa ttacctgtgg
    12781 tttcatttac tctaaacctg tgattcctct gaattatttt cattttaaag aaattgtatt
    12841 tgttaaatat gtactacaaa ctta
//
