LOCUS       sequence-6990-ex       12394 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tet-regulated (Tet-on) lentiviral vector for transgene (hEF-1alpha 
            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 12394)
  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 12394)
  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"; doi: "10"; journalName: 
            "Nat Methods"; date: "2006-02"; volume: "3"; issue: "2"; pages: 
            "109-16"
COMMENT     Sequence Label: pLVET-tTR-KRAB
FEATURES             Location/Qualifiers
     source          1..12394
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             2..635
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    682..807
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1300..1533
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1718..1762
                     /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"
     promoter        2120..3297
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2351..3288
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3245..3265
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    3345..3462
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      3539..3548
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3545..3547,3548..3550,3551..4261)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3545..3547,3548..3550,3551..4261)
                     /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(3590..3611)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3851..3870)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4198..4219
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4322..4889
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4340..4891
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4484..4501)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4711..4730
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4894..5514
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MARLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(4967..4986)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     CDS             5515..5535
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5638..5832
                     /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    5974..6562
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6027..6047)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6445..6456)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    join(6700..6718,6719..6741,6742..6760,6761..6783,6784..6802,
                     6803..6825,6826..6844,6845..6867,6868..6886,6887..6909,
                     6910..6928,6929..6951,6952..6970)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     LTR             7009..7189
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(7214..7232)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7215..7232)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(7522..7541)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(7755..7774)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7874..7896
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7934..7952)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8020..8124
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8125..8193,8194..8985)
                     /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(8343..8362)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9156..9744
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9645..9664
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9898..9915
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(9985..10005)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9990..10319
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10170..10305
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          11453..11518
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             11648..11668
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    12093..12227
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12130..12149)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12184..12203
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ttggaagggc taattcactc ccaaagaaga caagatatcc ttgatctgtg gatctaccac
       61 acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt cagatatcca
      121 ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt agaagaggcc
      181 aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat ggatgacccg
      241 gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca cgtggcccga
      301 gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa gggactttcc
      361 gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg agccctcaga
      421 tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag accagatctg
      481 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
      541 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
      601 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
      661 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
      721 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
      781 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
      841 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
      901 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
      961 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
     1021 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
     1081 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
     1141 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
     1201 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
     1261 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
     1321 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
     1381 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
     1441 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
     1501 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
     1561 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
     1621 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
     1681 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
     1741 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
     1801 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
     1861 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
     1921 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
     1981 agacagatcc attcgattag tgaacggatc tcgacggtat cgatgtcgac gataagcttt
     2041 gcaaagatgg ataaagtttt aaacagagag gaatctttgc agctaatgga ccttctaggt
     2101 cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca catcgcccac
     2161 agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg
     2221 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg
     2281 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc
     2341 cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg
     2401 cccttgcgtg ccttgaatta cttccactgg ctgcagtacg tgattcttga tcccgagctt
     2461 cgggttggaa gtgggtggga gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt
     2521 gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt
     2581 cgcgcctgtc tcgctgcttt cgataagtct ctagccattt aaaatttttg atgacctgct
     2641 gcgacgcttt ttttctggca agatagtctt gtaaatgcgg gccaagatct gcacactggt
     2701 atttcggttt ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg
     2761 gcgaggcggg gcctgcgagc gcggccaccg agaatcggac gggggtagtc tcaagctggc
     2821 cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg
     2881 ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca
     2941 gggagctcaa aatggaggac gcggcgctcg ggagagcggg cgggtgagtc acccacacaa
     3001 aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg actccacgga gtaccgggcg
     3061 ccgtccaggc acctcgatta gttctcgagc ttttggagta cgtcgtcttt aggttggggg
     3121 gaggggtttt atgcgatgga gtttccccac actgagtggg tggagactga agttaggcca
     3181 gcttggcact tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg atcttggttc
     3241 attctcaagc ctcagacagt ggttcaaagt ttttttcttc catttcaggt gtcgtgagga
     3301 atttcgacat ttaaatttaa ttaatctcga cggtatcggt taacttttaa aagaaaaggg
     3361 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     3421 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttatcgatca cgagactagc
     3481 ctcgaggttt aaactacggg atccaggcct aagcttacgc gtcctagcgc taccggtcgc
     3541 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     3601 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     3661 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     3721 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     3781 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     3841 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     3901 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     3961 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     4021 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     4081 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     4141 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     4201 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     4261 gtccggactc agatctcgac tagctagtag ctagctagct agtcgagctc aagcttcgaa
     4321 ttccgccccc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg
     4381 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa
     4441 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg
     4501 caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca
     4561 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc
     4621 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt
     4681 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg
     4741 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca
     4801 tgctttacat gtgtttagtc gaggttaaaa aacgtctagg ccccccgaac cacggggacg
     4861 tggttttcct ttgaaaaaca cgatgataat accatggcta gattagataa aagtaaagtg
     4921 attaacagcg cattagagct gcttaatgag gtcggaatcg aaggtttaac aacccgtaaa
     4981 ctcgcccaga agctaggtgt agagcagcct acattgtatt ggcatgtaaa aaataagcgg
     5041 gctttgctcg acgccttagc cattgagatg ttagataggc accatactca cttttgccct
     5101 ttagaagggg aaagctggca agatttttta cgtaataacg ctaaaagttt tagatgtgct
     5161 ttactaagtc atcgcgatgg agcaaaagta catttaggta cacggcctac agaaaaacag
     5221 tatgaaactc tcgaaaatca attagccttt ttatgccaac aaggtttttc actagagaat
     5281 gcattatatg cactcagcgc tgtggggcat tttactttag gttgcgtatt ggaagatcaa
     5341 gagcatcaag tcgctaaaga agaaagggaa acacctacta ctgatagtat gccgccatta
     5401 ttacgacaag ctatcgaatt atttgatcac caaggtgcag agccagcctt cttattcggc
     5461 cttgaattga tcatatgcgg attagaaaaa caacttaaat gtgaaagtgg gtcgccaaaa
     5521 aagaagagaa aggtcgacgg cggtggtgct ttgtctcctc agcactctgc tgtcactcaa
     5581 ggaagtatca tcaagaacaa ggagggcatg gatgctaagt cactaactgc ctggtcccgg
     5641 acactggtga ccttcaagga tgtatttgtg gacttcacca gggaggagtg gaagctgctg
     5701 gacactgctc agcagatcgt gtacagaaat gtgatgctgg agaactataa gaacctggtt
     5761 tccttgggtt atcagcttac taagccagat gtgatcctcc ggttggagaa gggagaagag
     5821 ccctggctgg tggagagaga aattcaccaa gagacccatc ctgattcaga gactgcattt
     5881 gaaatcaaat catcagttta aggatctcga ctagctagta gctagctagc tagtcgagct
     5941 caagcttcga attcccggga ctagtcatat gataatcaac ctctggatta caaaatttgt
     6001 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     6061 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     6121 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     6181 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     6241 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     6301 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     6361 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     6421 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     6481 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     6541 tccctttggg ccgcctcccc gcatcggtac gtatggccag gtacctttaa gaccaatgac
     6601 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     6661 aattcactcc caacgaagac aagatctcga gtttaccact ccctatcagt gatagagaaa
     6721 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac
     6781 cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata
     6841 gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga
     6901 gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca ctccctatca
     6961 gtgatagaga aaagtgaaag tcggatcgat ctgctttttg cttgtactgg gtctctctgg
     7021 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     7081 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     7141 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag catctagaat
     7201 taattccgtg tattctatag tgtcacctaa atcgtatgtg tatgatacat aaggttatgt
     7261 attaattgta gccgcgttct aacgacaata tgtacaagcc taattgtgta gcatctggct
     7321 tactgaagca gaccctatca tctctctcgt aaactgccgt cagagtcggt ttggttggac
     7381 gaaccttctg agtttctggt aacgccgtcc cgcacccgga aatggtcagc gaaccaatca
     7441 gcagggtcat cgctagccag atcctctacg ccggacgcat cgtggccggc atcaccggcg
     7501 ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc
     7561 gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg
     7621 ggggactgtt gggcgccatc tccttgcatg caccattcct tgcggcggcg gtgctcaacg
     7681 gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgtcgaa
     7741 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     7801 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     7861 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     7921 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     7981 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     8041 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     8101 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     8161 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     8221 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     8281 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     8341 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     8401 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     8461 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     8521 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     8581 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     8641 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     8701 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     8761 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     8821 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     8881 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     8941 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     9001 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     9061 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     9121 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     9181 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     9241 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     9301 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     9361 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     9421 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     9481 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     9541 cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     9601 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     9661 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     9721 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     9781 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     9841 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     9901 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     9961 tggccgattc attaatgcag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
    10021 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
    10081 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
    10141 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
    10201 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
    10261 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
    10321 agcttggaca caagacaggc ttgcgagata tgtttgagaa taccacttta tcccgcgtca
    10381 gggagaggca gtgcgtaaaa agacgcggac tcatgtgaaa tactggtttt tagtgcgcca
    10441 gatctctata atctcgcgca acctattttc ccctcgaaca ctttttaagc cgtagataaa
    10501 caggctggga cacttcacat gagcgaaaaa tacatcgtca cctgggacat gttgcagatc
    10561 catgcacgta aactcgcaag ccgactgatg ccttctgaac aatggaaagg cattattgcc
    10621 gtaagccgtg gcggtctgta ccgggtgcgt tactggcgcg tgaactgggt attcgtcatg
    10681 tcgataccgt ttgtatttcc agctacgatc acgacaacca gcgcgagctt aaagtgctga
    10741 aacgcgcaga aggcgatggc gaaggcttca tcgttattga tgacctggtg gataccggtg
    10801 gtactgcggt tgcgattcgt gaaatgtatc caaaagcgca ctttgtcacc atcttcgcaa
    10861 aaccggctgg tcgtccgctg gttgatgact atgttgttga tatcccgcaa gatacctgga
    10921 ttgaacagcc gtgggatatg ggcgtcgtat tcgtcccgcc aatctccggt cgctaatctt
    10981 ttcaacgcct ggcactgccg ggcgttgttc tttttaactt caggcgggtt acaatagttt
    11041 ccagtaagta ttctggaggc tgcatccatg acacaggcaa acctgagcga aaccctgttc
    11101 aaaccccgct ttaaacatcc tgaaacctcg acgctagtcc gccgctttaa tcacggcgca
    11161 caaccgcctg tgcagtcggc ccttgatggt aaaaccatcc ctcactggta tcgcatgatt
    11221 aaccgtctga tgtggatctg gcgcggcatt gacccacgcg aaatcctcga cgtccaggca
    11281 cgtattgtga tgagcgatgc cgaacgtacc gacgatgatt tatacgatac ggtgattggc
    11341 taccgtggcg gcaactggat ttatgagtgg gccccggatc tttgtgaagg aaccttactt
    11401 ctgtggtgtg acataattgg acaaactacc tacagagatt taaagctcta aggtaaatat
    11461 aaaattttta agtgtataat gtgttaaact actgattcta attgtttgtg tattttagat
    11521 tccaacctat ggaactgatg aatgggagca gtggtggaat gcctttaatg aggaaaacct
    11581 gttttgctca gaagaaatgc catctagtga tgatgaggct actgctgact ctcaacattc
    11641 tactcctcca aaaaagaaga gaaaggtaga agaccccaag gactttcctt cagaattgct
    11701 aagttttttg agtcatgctg tgtttagtaa tagaactctt gcttgctttg ctatttacac
    11761 cacaaaggaa aaagctgcac tgctatacaa gaaaattatg gaaaaatatt ctgtaacctt
    11821 tataagtagg cataacagtt ataatcataa catactgttt tttcttactc cacacaggca
    11881 tagagtgtct gctattaata actatgctca aaaattgtgt acctttagct ttttaatttg
    11941 taaaggggtt aataaggaat atttgatgta tagtgccttg actagagatc ataatcagcc
    12001 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
    12061 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
    12121 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
    12181 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcaac tggataactc
    12241 aagctaacca aaatcatccc aaacttccca ccccataccc tattaccact gccaattacc
    12301 tagtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat
    12361 tgtatttgtt aaatatgtac tacaaactta gtag
//
