LOCUS       sequence-228571-       10621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral TET-ON inducible GFP-Progerin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10621)
  AUTHORS   Kubben N, Brimacombe KR, Donegan M, Li Z, Misteli T
  TITLE     A high-content imaging-based screening pipeline for the systematic 
            identification of anti-progeroid compounds.
  JOURNAL   Methods. 2015 Sep 1. pii: S1046-2023(15)30070-0. doi: 
            10.1016/j.ymeth.2015.08.024.
  PUBMED    26341717
REFERENCE   2  (bases 1 to 10621)
  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: "Methods. 
            2015 Sep 1. pii: S1046-2023(15)30070-0. doi: 
            10.1016/j.ymeth.2015.08.024."
COMMENT     Sequence Label: pLenti CMV TRE3G Neo GFP-Progerin
FEATURES             Location/Qualifiers
     source          1..10621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     98..115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    232..253
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(268..273,274..291,292..298)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    306..322
                     /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     311..333
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     330..346
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     330..346
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     365..385
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        367..385
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        413..639
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             640..820
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    867..992
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1485..1718
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1903..1947
                     /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    2214..2331
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2358..2736
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    2366..2384
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2402..2420
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2438..2456
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2474..2492
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2510..2528
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2546..2564
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2582..2600
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2645..2669
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2782..2806
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2820..2829
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2826..2828,2829..2831,2832..3542)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2826..2828,2829..2831,2832..3542)
                     /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(2871..2892)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3132..3151)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3479..3500
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(5459..5483)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    5514..6102
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5567..5587)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5985..5996)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        6145..6644
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(6180..6203)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6577..6596
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             6658..7461
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6721..6740)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7331..7350
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             7552..7785
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7857..7978
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7894..7913)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7948..7967
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      8018..8153
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(8173..8192)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8174..8192)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8202..8219)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8202..8218)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8211..8233)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      8360..8815
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8447..8466)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8657..8678
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8841..8945
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8946..9014,9015..9806)
                     /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(9164..9183)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9977..10565
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10466..10485
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
       61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
      181 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
      241 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
      301 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
      361 gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct
      421 tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa
      481 ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta
      541 ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc
      601 agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag
      661 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
      721 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
      781 tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact
      841 tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc
      901 gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag
      961 gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc
     1021 gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag
     1081 tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag
     1141 aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac
     1201 ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa
     1261 aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg
     1321 cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga
     1381 agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag
     1441 gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt
     1501 gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag
     1561 gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag
     1621 gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc
     1681 ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga
     1741 aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa
     1801 cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc
     1861 ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta
     1921 ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg
     1981 tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct
     2041 gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac
     2101 ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga
     2161 gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt aacttttaaa
     2221 agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca
     2281 gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tatcgatcac
     2341 gagactagcc tcgagttggc tttactccct atcagtgata gagaacgtat gaagagttta
     2401 ctccctatca gtgatagaga acgtatgcag actttactcc ctatcagtga tagagaacgt
     2461 ataaggagtt tactccctat cagtgataga gaacgtatga ccagtttact ccctatcagt
     2521 gatagagaac gtatctacag tttactccct atcagtgata gagaacgtat atccagttta
     2581 ctccctatca gtgatagaga acgtataagc tttaggcgtg tacggtgggc gcctataaaa
     2641 gcagagctcg tttagtgaac cgtcagatcg cctggagcaa ttccacaaca cttttgtctt
     2701 ataccaactt tccgtaccac ttcctaccct cgtaaaccgg ttcgtcgact agtccagtgt
     2761 ggtggaattc tgcagatatc aacaagtttg tacaaaaaag caggctttaa ataccggtcg
     2821 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     2881 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     2941 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     3001 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     3061 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     3121 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     3181 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     3241 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     3301 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     3361 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     3421 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     3481 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     3541 agtccggact cagatctcga gatttcgaaa aaggcgccgg gtcgaccgag accccgtccc
     3601 agcggcgcgc cacccgcagc ggggcgcagg ccagctccac tccgctgtcg cccacccgca
     3661 tcacccggct gcaggagaag gaggacctgc aggagctcaa tgatcgcttg gcggtctaca
     3721 tcgaccgtgt gcgctcgctg gaaacggaga acgcagggct gcgccttcgc atcaccgagt
     3781 ctgaagaggt ggtcagccgc gaggtgtccg gcatcaaggc cgcctacgag gccgagctcg
     3841 gggatgcccg caagaccctt gactcagtag ccaaggagcg cgcccgcctg cagctggagc
     3901 tgagcaaagt gcgtgaggag tttaaggagc tgaaagcgcg caataccaag aaggagggtg
     3961 acctgatagc tgctcaggct cggctgaagg acctggaggc tctgctgaac tccaaggagg
     4021 ccgcactgag cactgctctc agtgagaagc gcacgctgga gggcgagctg catgatctgc
     4081 ggggccaggt ggccaagctt gaggcagccc taggtgaggc caagaagcaa cttcaggatg
     4141 agatgctgcg gcgggtggat gctgagaaca ggctgcagac catgaaggag gaactggact
     4201 tccagaagaa catctacagt gaggagctgc gtgagaccaa gcgccgtcat gagacccgac
     4261 tggtggagat tgacaatggg aagcagcgtg agtttgagag ccggctggcg gatgcgctgc
     4321 aggaactgcg ggcccagcat gaggaccagg tggagcagta taagaaggag ctggagaaga
     4381 cttattctgc caagctggac aatgccaggc agtctgctga gaggaacagc aacctggtgg
     4441 gggctgccca cgaggagctg cagcagtcgc gcatccgcat cgacagcctc tctgcccagc
     4501 tcagccagct ccagaagcag ctggcagcca aggaggcgaa gcttcgagac ctggaggact
     4561 cactggcccg tgagcgggac accagccggc ggctgctggc ggaaaaggag cgggagatgg
     4621 ccgagatgcg ggcaaggatg cagcagcagc tggacgagta ccaggagctt ctggacatca
     4681 agctggccct ggacatggag atccacgcct accgcaagct cttggagggc gaggaggaga
     4741 ggctacgcct gtcccccagc cctacctcgc agcgcagccg tggccgtgct tcctctcact
     4801 catcccagac acagggtggg ggcagcgtca ccaaaaagcg caaactggag tccactgaga
     4861 gccgcagcag cttctcacag cacgcacgca ctagcgggcg cgtggccgtg gaggaggtgg
     4921 atgaggaggg caagtttgtc cggctgcgca acaagtccaa tgaggaccag tccatgggca
     4981 attggcagat caagcgccag aatggagatg atcccttgct gacttaccgg ttcccaccaa
     5041 agttcaccct gaaggctggg caggtggtga cgatctgggc tgcaggagct ggggccaccc
     5101 acagcccccc taccgacctg gtgtggaagg cacagaacac ctggggctgc gggaacagcc
     5161 tgcgtacggc tctcatcaac tccactgggg aagaagtggc catgcgcaag ctggtgcgct
     5221 cagtgactgt ggttgaggac gacgaggatg aggatggaga tgacctgctc catcaccacc
     5281 acggctccca ctgcagcagc tcgggggacc ccgctgagta caacctgcgc tcgcgcaccg
     5341 tgctgtgcgg gacctgcggg cagcctgccg acaaggcatc tgccagcggc tcaggagccc
     5401 agagccccca gaactgcagc atcatgtaag aattcgcggc cgcactcgag atatctagac
     5461 ccagctttct tgtacaaagt ggttgatatc cagcacagtg gcggccgctc gacaatcaac
     5521 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     5581 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     5641 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     5701 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     5761 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     5821 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     5881 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
     5941 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     6001 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     6061 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt ctgcagatat
     6121 ccggactagt gatctaattc taccgggtag gggaggcgct tttcccaagg cagtctggag
     6181 catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca
     6241 cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc
     6301 caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca
     6361 ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg
     6421 agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt
     6481 tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg
     6541 gcggggcggg cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca
     6601 cgtctgccgc gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agccaatatg
     6661 ggatcggcca ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     6721 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     6781 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     6841 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     6901 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     6961 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     7021 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     7081 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     7141 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     7201 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     7261 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     7321 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     7381 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     7441 cttcttgacg agttcttctg aggggatccg tcgagggccc ggtaccttta agaccaatga
     7501 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     7561 tagctcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag
     7621 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     7681 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     7741 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
     7801 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact
     7861 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     7921 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     7981 atgtctggct ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     8041 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     8101 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagggac
     8161 gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa
     8221 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     8281 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     8341 agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     8401 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     8461 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     8521 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     8581 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     8641 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     8701 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     8761 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     8821 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     8881 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     8941 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     9001 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     9061 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     9121 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     9181 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     9241 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     9301 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     9361 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     9421 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     9481 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     9541 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     9601 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     9661 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     9721 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     9781 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     9841 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     9901 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     9961 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
    10021 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
    10081 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
    10141 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
    10201 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
    10261 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
    10321 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
    10381 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
    10441 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
    10501 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
    10561 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
    10621 a
//
